CN107164394B - Biosynthetic gene cluster of atypical keratinocyte compound nenestatin A and application thereof - Google Patents

Abstract

The invention discloses a biosynthetic gene cluster of atypical keratinocyte compound nenostatin A and application thereof. The nucleotide sequence of the biosynthetic gene cluster of the atypical angulatin compound nenestatin A is shown as the base sequence of 3153-68576 site of SEQ ID NO. 1. The gene and protein information related to the biosynthesis of the nerestatin A provided by the invention can help people to understand the biosynthesis mechanism of the natural product of the ceratin, and provide materials and theoretical basis for further genetic modification. The gene and protein provided by the invention can be used for searching and discovering the nenestatin A compound, gene or protein which can be used for medicine, health or agriculture.

Description

Translated fromChinese

一种非典型角环素类化合物nenestatin A的生物合成基因簇及其应用Biosynthetic gene cluster of an atypical keratin-like compound nenestatin Aand its application

技术领域：Technical field:

本发明属于微生物基因工程领域，具体涉及一种非典型角环素类化合物nenestatin A的生物合成基因簇及其应用。The invention belongs to the field of microbial genetic engineering, and in particular relates to a biosynthetic gene cluster of an atypical keratin compound nenestatin A and its application.

背景技术：Background technique:

Nenestatin A属于非典型角环素类化合物，由小单孢菌(M.echinospora)SCSIO04089产生。近年来蓬勃发展起来的组合生物合成技术，为改造nenestatin A的生产菌SCSIO 04089带来了新的机遇。在学习自然界非典型角环素类化合物生物合成机理的基础上，采用组合生物合成技术对化合物的生物合成基因、调控基因进行体内敲除、突变、置换和重组等操作，不但能够生产“非天然”的天然产物结构类似物，而且还可以提高天然产物的产量，或定向积累所需要的天然产物，为天然产物的发现和药物开发提供化合物和生物活性多样化。迄今为止，有关非典型角环素类化合物nenestatin A的生物合成基因簇、氧化酶NES27等生物合成关键酶的生物催化机理及应用方面的研究，国内外尚未研究清楚。Nenestatin A belongs to atypical keratin-type compounds, produced by M. echinospora SCSIO04089. The combinatorial biosynthesis technology that has developed vigorously in recent years has brought new opportunities for the transformation of nenestatin A-producing strain SCSIO 04089. On the basis of studying the biosynthesis mechanism of atypical keratin-type compounds in nature, the combined biosynthesis technology is used to carry out in vivo knockout, mutation, replacement and recombination of the biosynthetic genes and regulatory genes of the compounds, which can not only produce "non-natural" "Structural analogs of natural products, but also can improve the yield of natural products, or directional accumulation of natural products required, providing compounds and biological activity diversification for natural product discovery and drug development. So far, the research on the biosynthesis gene cluster of the atypical keratin-like compound nenestatin A and the biocatalytic mechanism and application of key biosynthetic enzymes such as the oxidase NES27 has not been clearly studied at home and abroad.

发明内容：Invention content:

本发明的目的是提供一种非典型角环素类化合物nenestatin A的生物合成基因簇及其应用。The purpose of the present invention is to provide a biosynthetic gene cluster of an atypical keratin compound nenestatin A and its application.

本发明是以南海北部深海沉积物来源小单孢菌(M.echinospora)SCSIO 04089中非典型角环素类化合物nenestatin A为目标分子，在筛选到生物合成基因簇的基础上，综合运用分子生物学、微生物学、合成生物学、生物化学以及有机化学相结合的技术，阐述nenestatin A的生物合成机制，为利用组合生物合成方法修饰改造nenestatin A类化合物提供依据，为药物筛选提供化合物实体。The present invention takes the atypical keratin-type compound nenestatin A from M. echinospora SCSIO 04089 from deep-sea sediments in the northern South China Sea as a target molecule, and on the basis of screening biosynthetic gene clusters, comprehensively utilizes molecular biology Using the combination of science, microbiology, synthetic biology, biochemistry and organic chemistry, the biosynthesis mechanism of nenestatin A is described, which provides a basis for the modification and transformation of nenestatin A compounds by combinatorial biosynthesis, and provides compound entities for drug screening.

本发明的第一个目的是提供非典型角环素类化合物nenestatin A的生物合成基因簇，来源于南海北部深海沉积物小单孢菌(M.echinospora)SCSIO 04089，该生物合成基因簇的核苷酸序列如SEQ ID NO.1的第3153-68576位的碱基序列所示，包含60个基因，具体为：The first object of the present invention is to provide a biosynthetic gene cluster of an atypical keratinoid compound nenestatin A, which is derived from M. echinospora SCSIO 04089 in the deep-sea sediments of the northern South China Sea. The core of the biosynthetic gene cluster is The nucleotide sequence is shown in the nucleotide sequence of positions 3153-68576 of SEQ ID NO.1, including 60 genes, specifically:

1)nes1位于基因簇核苷酸序列3153-3635个碱基处，长度为483bp，编码未知功能蛋白，160个氨基酸。1) nes1 is located at 3153-3635 bases in the nucleotide sequence of the gene cluster, with a length of 483 bp, and encodes a protein of unknown function with 160 amino acids.

2)nes2位于基因簇核苷酸序列3788-4618个碱基处，长度为831bp，编码ABC转运蛋白，276个氨基酸。2) nes2 is located at 3788-4618 bases in the nucleotide sequence of the gene cluster, with a length of 831 bp, and encodes an ABC transporter with 276 amino acids.

3)nes3位于基因簇核苷酸序列4615-5574个碱基处，长度为960bp，编码ABC转运蛋白，319个氨基酸。3) nes3 is located at 4615-5574 bases in the nucleotide sequence of the gene cluster, with a length of 960 bp, encoding an ABC transporter, with 319 amino acids.

4)nes4位于基因簇核苷酸序列5670-6095个碱基处，长度为426bp，编码醛酮变位酶/博来霉素抗性蛋白/双加氧酶，141个氨基酸。4) nes4 is located at 5670-6095 bases of the nucleotide sequence of the gene cluster, with a length of 426 bp, encoding aldosterone mutase/bleomycin resistance protein/dioxygenase, with 141 amino acids.

5)nes5位于基因簇核苷酸序列6215-6991个碱基处，长度为777bp，编码推测的蛋白，258个氨基酸。5) nes5 is located at 6215-6991 bases of the nucleotide sequence of the gene cluster, with a length of 777 bp, and encodes a deduced protein with 258 amino acids.

6)nes6位于基因簇核苷酸序列7114-7926个碱基处，长度为813bp，编码SARP家族调控蛋白，270个氨基酸。6) nes6 is located at 7114-7926 bases in the nucleotide sequence of the gene cluster, with a length of 813 bp, encoding a regulatory protein of the SARP family, with 270 amino acids.

7)nes7位于基因簇核苷酸序列9201-10049个碱基处，长度为849bp，编码AraC家族的转录调控因子，282个氨基酸。7) nes7 is located at 9201-10049 bases in the nucleotide sequence of the gene cluster, with a length of 849 bp, and encodes a transcriptional regulator of the AraC family, with 282 amino acids.

8)nes8位于基因簇核苷酸序列10479-11873个碱基处，长度为1395bp，编码AraC家族的转录调控因子，464个氨基酸。8) nes8 is located at 10479-11873 bases in the nucleotide sequence of the gene cluster, with a length of 1395 bp, and encodes a transcriptional regulator of the AraC family, with 464 amino acids.

9)nes9位于基因簇核苷酸序列11953-12618个碱基处，长度为666bp，编码NDP-己糖2,3脱水酶，221个氨基酸。9) nes9 is located at 11953-12618 bases in the nucleotide sequence of the gene cluster, with a length of 666 bp, encoding NDP-hexose 2,3 dehydratase, with 221 amino acids.

10)nes10位于基因簇核苷酸序列12655-13101个碱基处，长度为447bp，编码羧酸粘康酸内酯脱羧酶，148个氨基酸。10) nes10 is located at 12655-13101 bases of the nucleotide sequence of the gene cluster, with a length of 447 bp, encoding a carboxymuconolide decarboxylase, with 148 amino acids.

11)nes11位于基因簇核苷酸序列13171-14025个碱基处，长度为855bp，编码谷氨酰基水解酶/氨基转移酶，284个氨基酸。11) nes11 is located at 13171-14025 bases of the nucleotide sequence of the gene cluster, with a length of 855 bp, encoding glutamyl hydrolase/aminotransferase, with 284 amino acids.

12)nes12位于基因簇核苷酸序列14542-16191个碱基处，长度为1650bp，编码FAD依赖的单加氧酶，549个氨基酸。12) nes12 is located at 14542-16191 bases in the nucleotide sequence of the gene cluster, with a length of 1650 bp, encoding a FAD-dependent monooxygenase, with 549 amino acids.

13)nes13位于基因簇核苷酸序列16329-17798个碱基处，长度为1470bp，编码原卟啉氧化酶，489个氨基酸。13) nes13 is located at 16329-17798 bases in the nucleotide sequence of the gene cluster, with a length of 1470 bp, encoding protoporphyrin oxidase, with 489 amino acids.

14)nes14位于基因簇核苷酸序列17907-18959个碱基处，长度为1053bp，编码核酸酶，350个氨基酸。14) nes14 is located at 17907-18959 bases of the nucleotide sequence of the gene cluster, with a length of 1053 bp, and encodes a nuclease with 350 amino acids.

15)nes15位于基因簇核苷酸序列19096-20577个碱基处，长度为1482bp，编码FAD依赖的单加氧酶，493个氨基酸。15) nes15 is located at 19096-20577 bases in the nucleotide sequence of the gene cluster, with a length of 1482 bp, encoding a FAD-dependent monooxygenase, with 493 amino acids.

16)nes16位于基因簇核苷酸序列20574-22034个碱基处，长度为1461bp，编码FAD依赖的单加氧酶，486个氨基酸。16) nes16 is located at 20574-22034 bases in the nucleotide sequence of the gene cluster, with a length of 1461 bp, and encodes a FAD-dependent monooxygenase with 486 amino acids.

17)nes17位于基因簇核苷酸序列22272-23000个碱基处，长度为729bp，编码短链脱氢酶/还原酶，242个氨基酸。17) nes17 is located at 22272-23000 bases of the nucleotide sequence of the gene cluster, with a length of 729 bp, encoding a short-chain dehydrogenase/reductase, with 242 amino acids.

18)nes18位于基因簇核苷酸序列23075-23920个碱基处，长度为846bp，编码NmrA家族蛋白，281个氨基酸。18) nes18 is located at 23075-23920 bases in the nucleotide sequence of the gene cluster, with a length of 846 bp, and encodes a NmrA family protein with 281 amino acids.

19)nes19位于基因簇核苷酸序列23983-24600个碱基处，长度为618bp，编码DSBA氧化还原酶，205个氨基酸。19) nes19 is located at 23983-24600 bases of the nucleotide sequence of the gene cluster, with a length of 618 bp, encoding DSBA oxidoreductase, with 205 amino acids.

20)nes20位于基因簇核苷酸序列24663-24992个碱基处，长度为330bp，编码聚酮合成酶/环化酶，109个氨基酸。20) nes20 is located at 24663-24992 bases of the nucleotide sequence of the gene cluster, with a length of 330 bp, encoding a polyketide synthase/cyclase, with 109 amino acids.

21)nes21位于基因簇核苷酸序列25036-25821个碱基处，长度为786bp，编码聚酮合成，酮基还原酶，261个氨基酸。21) nes21 is located at 25036-25821 bases of the nucleotide sequence of the gene cluster, with a length of 786 bp, encoding polyketide synthesis, ketoreductase, and 261 amino acids.

22)nes22位于基因簇核苷酸序列26017-27054个碱基处，长度为1038bp，编码氧甲基转移酶，345个氨基酸。22) nes22 is located at 26017-27054 bases in the nucleotide sequence of the gene cluster, with a length of 1038 bp, encoding an oxymethyltransferase, with 345 amino acids.

23)nes23位于基因簇核苷酸序列27109-28620个碱基处，长度为1512bp，编码FAD依赖的单加氧酶，503个氨基酸。23) nes23 is located at 27109-28620 bases of the nucleotide sequence of the gene cluster, with a length of 1512 bp, encoding a FAD-dependent monooxygenase, with 503 amino acids.

24)nes24位于基因簇核苷酸序列28647-29606个碱基处，长度为960bp，编码聚酮合成，酮基还原酶，319个氨基酸。24) nes24 is located at 28647-29606 bases of the nucleotide sequence of the gene cluster, with a length of 960bp, encoding polyketide synthesis, ketoreductase, and 319 amino acids.

25)nes25位于基因簇核苷酸序列29603-31039个碱基处，长度为1437bp，编码FAD依赖的单加氧酶，478个氨基酸。25) nes25 is located at 29603-31039 bases of the nucleotide sequence of the gene cluster, with a length of 1437 bp, encoding a FAD-dependent monooxygenase, with 478 amino acids.

26)nes26位于基因簇核苷酸序列31026-32504个碱基处，长度为1479bp，编码FAD依赖的单加氧酶，492个氨基酸。26) nes26 is located at 31026-32504 bases of the nucleotide sequence of the gene cluster, with a length of 1479 bp, encoding a FAD-dependent monooxygenase, with 492 amino acids.

27)nes27位于基因簇核苷酸序列32561-33304个碱基处，长度为744bp，编码蒽酮氧化酶，247个氨基酸。27) nes27 is located at 32561-33304 bases of the nucleotide sequence of the gene cluster, with a length of 744 bp, encoding anthrone oxidase, with 247 amino acids.

28)nes28位于基因簇核苷酸序列33373-35256个碱基处，长度为1884bp，编码未知功能蛋白，627个氨基酸。28) nes28 is located at 33373-35256 bases in the nucleotide sequence of the gene cluster, with a length of 1884 bp, and encodes a protein of unknown function with 627 amino acids.

29)nes29位于基因簇核苷酸序列35237-35593个碱基处，长度为357bp，编码4Fe-4S铁氧还蛋白，118个氨基酸。29) nes29 is located at 35237-35593 bases of the nucleotide sequence of the gene cluster, with a length of 357bp, encoding 4Fe-4S ferredoxin, with 118 amino acids.

30)nes30位于基因簇核苷酸序列35668-36462个碱基处，长度为795bp，编码未知功能蛋白，264个氨基酸。30) nes30 is located at 35668-36462 bases of the nucleotide sequence of the gene cluster, with a length of 795 bp, encoding a protein of unknown function, with 264 amino acids.

31)nes31位于基因簇核苷酸序列36499-37992个碱基处，长度为1494bp，编码谷氨酰胺合成酶，497个氨基酸。31) nes31 is located at 36499-37992 bases of the nucleotide sequence of the gene cluster, with a length of 1494 bp, encoding glutamine synthase, with 497 amino acids.

32)nes32位于基因簇核苷酸序列37995-39413个碱基处，长度为1419bp，编码酰胺酶，472个氨基酸。32) nes32 is located at 37995-39413 bases of the nucleotide sequence of the gene cluster, with a length of 1419 bp, encoding amidase, with 472 amino acids.

33)nes33位于基因簇核苷酸序列39624-40910个碱基处，长度为1287bp，编码腺苷酸琥珀酸裂合酶，428个氨基酸。33) nes33 is located at 39624-40910 bases of the nucleotide sequence of the gene cluster, with a length of 1287 bp, encoding adenylate succinate lyase, with 428 amino acids.

34)nes34位于基因簇核苷酸序列40894-41295个碱基处，长度为402bp，编码N-乙酰基转移酶，133个氨基酸。34) nes34 is located at 40894-41295 bases of the nucleotide sequence of the gene cluster, with a length of 402 bp, encoding N-acetyltransferase, with 133 amino acids.

35)nes35位于基因簇核苷酸序列41406-43172个碱基处，长度为1767bp，编码ABC转运蛋白，588个氨基酸。35) nes35 is located at 41406-43172 bases in the nucleotide sequence of the gene cluster, with a length of 1767 bp, encoding an ABC transporter, with 588 amino acids.

36)nes36位于基因簇核苷酸序列43575-43904个碱基处，长度为330bp，编码Rieske型非血红素铁加氧酶，109个氨基酸。36) nes36 is located at 43575-43904 bases of the nucleotide sequence of the gene cluster, with a length of 330 bp, encoding a Rieske type non-heme iron oxygenase, with 109 amino acids.

37)nes37位于基因簇核苷酸序列44148-44558个碱基处，长度为411bp，编码醛酮变位酶，136个氨基酸。37) nes37 is located at 44148-44558 bases of the nucleotide sequence of the gene cluster, with a length of 411 bp, encoding an aldosterone mutase, with 136 amino acids.

38)nes38位于基因簇核苷酸序列44642-45442个碱基处，长度为801bp，编码醛酮变位酶，266个氨基酸。38) nes38 is located at 44642-45442 bases of the nucleotide sequence of the gene cluster, with a length of 801 bp, encoding an aldosterone mutase, and having 266 amino acids.

39)nes39位于基因簇核苷酸序列45500-47050个碱基处，长度为1551bp，编码分泌肽酶，516个氨基酸。39) nes39 is located at 45500-47050 bases of the nucleotide sequence of the gene cluster, with a length of 1551 bp, encoding a secreted peptidase, with 516 amino acids.

40)nes40位于基因簇核苷酸序列47571-48692个碱基处，长度为1122bp，编码糖基转移酶，373个氨基酸。40) nes40 is located at 47571-48692 bases of the nucleotide sequence of the gene cluster, with a length of 1122 bp, and encodes a glycosyltransferase with 373 amino acids.

41)nes41位于基因簇核苷酸序列48689-49708个碱基处，长度为1020bp，编码氧甲基转移酶(糖合成)，339个氨基酸。41) nes41 is located at 48689-49708 bases in the nucleotide sequence of the gene cluster, with a length of 1020 bp, encoding an oxymethyltransferase (sugar synthesis), with 339 amino acids.

42)nes42位于基因簇核苷酸序列50074-51006个碱基处，长度为933bp，编码氮甲基转移酶(糖合成)，310个氨基酸。42) nes42 is located at 50074-51006 bases of the nucleotide sequence of the gene cluster, with a length of 933 bp, encoding nitrogen methyltransferase (sugar synthesis), 310 amino acids.

43)nes43位于基因簇核苷酸序列51054-52055个碱基处，长度为1002bp，编码NAD依赖的差向异构酶/脱水酶(糖合成)，333个氨基酸。43) nes43 is located at 51054-52055 bases of the nucleotide sequence of the gene cluster, with a length of 1002 bp, encoding an NAD-dependent epimerase/dehydratase (sugar synthesis), 333 amino acids.

44)nes44位于基因簇核苷酸序列52042-52767个碱基处，长度为726bp，编码甲基转移酶(糖合成)，241个氨基酸。44) nes44 is located at 52042-52767 bases in the nucleotide sequence of the gene cluster, with a length of 726 bp, encoding a methyltransferase (sugar synthesis), with 241 amino acids.

45)nes45位于基因簇核苷酸序列52764-53873个碱基处，长度为1110bp，编码氨基转移酶(糖合成)，369个氨基酸。45) nes45 is located at 52764-53873 bases in the nucleotide sequence of the gene cluster, with a length of 1110 bp, encoding an aminotransferase (sugar synthesis), with 369 amino acids.

46)nes46位于基因簇核苷酸序列53897-54508个碱基处，长度为612bp，编码NDP-4酮-6-脱氧己糖3,5-差向异构酶(糖合成)，203个氨基酸。46) nes46 is located at 53897-54508 bases of the gene cluster nucleotide sequence, with a length of 612bp, encoding NDP-4 keto-6-deoxyhexose 3,5-epimerase (sugar synthesis), 203 amino acids .

47)nes47位于基因簇核苷酸序列54685-55680个碱基处，长度为996bp，编码NDP-己糖-3-酮基还原酶(糖合成)，331个氨基酸。47) nes47 is located at 54685-55680 bases of the gene cluster nucleotide sequence, with a length of 996 bp, encoding NDP-hexose-3-keto reductase (sugar synthesis), 331 amino acids.

48)nes48位于基因簇核苷酸序列55735-56451个碱基处，长度为717bp，编码推测的蛋白，238个氨基酸。48) nes48 is located at 55735-56451 bases of the nucleotide sequence of the gene cluster, with a length of 717 bp, and encodes a deduced protein with 238 amino acids.

49)nes49位于基因簇核苷酸序列56544-57596个碱基处，长度为1053bp，编码糖基转移酶，350个氨基酸。49) nes49 is located at 56544-57596 bases of the nucleotide sequence of the gene cluster, with a length of 1053 bp, encoding a glycosyltransferase, with 350 amino acids.

50)nes50位于基因簇核苷酸序列57789-59009个碱基处，长度为1221bp，编码脱氧糖氨基转移酶(糖合成)，406个氨基酸。50) nes50 is located at 57789-59009 bases of the nucleotide sequence of the gene cluster, with a length of 1221 bp, encoding a deoxyglycosylaminotransferase (sugar synthesis), with 406 amino acids.

51)nes51位于基因簇核苷酸序列59205-60278个碱基处，长度为1074bp，编码dTDP-葡萄糖4，6脱水酶(糖合成)，357个氨基酸。51) nes51 is located at the nucleotide sequence of 59205-60278 bases in the gene cluster, with a length of 1074 bp, encoding dTDP-glucose 4,6 dehydratase (sugar synthesis), 357 amino acids.

52)nes52位于基因簇核苷酸序列60275-61546个碱基处，长度为1272bp，编码聚酮合成酶，423个氨基酸。52) nes52 is located at 60275-61546 bases of the nucleotide sequence of the gene cluster, with a length of 1272 bp, encoding a polyketide synthase, with 423 amino acids.

53)nes53位于基因簇核苷酸序列61543-62841个碱基处，长度为1299bp，编码聚酮链长因子，442个氨基酸。53) nes53 is located at 61543-62841 bases of the nucleotide sequence of the gene cluster, with a length of 1299 bp, encoding a polyketide chain length factor, with 442 amino acids.

54)nes54位于基因簇核苷酸序列62829-63083个碱基处，长度为255bp，编码酰基转运蛋白，84个氨基酸。54) nes54 is located at 62829-63083 bases of the nucleotide sequence of the gene cluster, with a length of 255 bp, encoding an acyl transporter, with 84 amino acids.

55)nes55位于基因簇核苷酸序列63065-64102个碱基处，长度为1038bp，编码酰基转移酶，345个氨基酸。55) nes55 is located at 63065-64102 bases of the nucleotide sequence of the gene cluster, with a length of 1038 bp, encoding an acyltransferase, with 345 amino acids.

56)nes56位于基因簇核苷酸序列64219-65196个碱基处，长度为978bp，编码酰基转移酶，325个氨基酸。56) nes56 is located at 64219-65196 bases of the nucleotide sequence of the gene cluster, with a length of 978 bp, encoding an acyltransferase, with 325 amino acids.

57)nes57位于基因簇核苷酸序列65225-65554个碱基处，长度为330bp，编码酰基载体蛋白，109个氨基酸。57) nes57 is located at 65225-65554 bases of the nucleotide sequence of the gene cluster, with a length of 330 bp, and encodes an acyl carrier protein with 109 amino acids.

58)nes58位于基因簇核苷酸序列65681-66136个碱基处，长度为456bp，编码转录调控因子，151个氨基酸。58) nes58 is located at 65681-66136 bases in the nucleotide sequence of the gene cluster, with a length of 456 bp, encoding a transcriptional regulator, with 151 amino acids.

59)nes59位于基因簇核苷酸序列66215-67639个碱基处，长度为1425bp，编码丙酰CoA羧化酶β亚基，474个氨基酸。59) nes59 is located at 66215-67639 bases in the nucleotide sequence of the gene cluster, with a length of 1425 bp, encoding the beta subunit of propionyl CoA carboxylase, with 474 amino acids.

60)nes60位于基因簇核苷酸序列67704-68576个碱基处，长度为873bp，编码dTDP-1-葡萄糖合成酶，290个氨基酸。60) nes60 is located at 67704-68576 bases of the gene cluster nucleotide sequence, with a length of 873 bp, encoding dTDP-1-glucose synthase, 290 amino acids.

SEQ ID NO.1所示序列的第3153-68576位核苷酸序列的互补序列可根据DNA碱基互补原则随时得到，且第3153-68576位的核苷酸序列或部分核苷酸序列可以通过聚合酶链式反应(PCR)或用合适的限制性内切酶酶切相应DNA或DNA重组技术获得。本发明提供了构建至少包含部分SEQ ID NO.1所示序列的第3153-68576位中DNA片段的重组DNA载体途径。The complementary sequence of the 3153-68576 nucleotide sequence of the sequence shown in SEQ ID NO.1 can be obtained at any time according to the principle of DNA base complementarity, and the 3153-68576 nucleotide sequence or part of the nucleotide sequence can be obtained by It can be obtained by polymerase chain reaction (PCR) or by appropriate restriction endonuclease digestion of the corresponding DNA or DNA recombination techniques. The present invention provides a method for constructing a recombinant DNA vector comprising at least part of the DNA fragment in positions 3153-68576 of the sequence shown in SEQ ID NO.1.

本发明还提供了nenestatin A生物合成基因被中断或其他基因改造的途径，至少其中之一的基因包含有SEQ ID NO.1的第3153-68576位中的核苷酸序列。The present invention also provides a pathway for the nenestatin A biosynthesis gene to be interrupted or other genetically modified, at least one of the genes comprising the nucleotide sequence in positions 3153-68576 of SEQ ID NO.1.

本发明所提供的核苷酸序列或部分核苷酸序列，可使用PCR探针法等技术，从其他生物体重筛选获得nenestatin A生物合成基因簇同源基因。For the nucleotide sequence or part of the nucleotide sequence provided by the present invention, the homologous gene of the nenestatin A biosynthesis gene cluster can be obtained by re-screening from other organisms by using techniques such as PCR probe method.

本发明所提供的核苷酸序列或部分核苷酸序列，可用于M.echinospora SCSIO04089基因组文库中定位更多的文库质粒。这些文库质粒至少包含本发明中的部分序列，也包含M.echinospora SCSIO 04089基因组中邻近区域未克隆DNA。The nucleotide sequence or part of the nucleotide sequence provided by the present invention can be used to locate more library plasmids in the M. echinospora SCSIO04089 genomic library. These library plasmids contain at least part of the sequences of the present invention, and also contain uncloned DNA from adjacent regions in the M. echinospora SCSIO 04089 genome.

包含本发明所提供的核苷酸序列或至少部分核苷酸序列的DNA片段，可以被体内外突变等修饰，包括插入、置换、缺失、易错聚合酶链式反应、位点特异性突变、不同序列重组、定向进化等。DNA fragments comprising the nucleotide sequences provided by the present invention or at least part of the nucleotide sequences can be modified by in vitro and in vivo mutations, including insertion, substitution, deletion, error-prone polymerase chain reaction, site-specific mutation, Recombination of different sequences, directed evolution, etc.

包含本发明所提供的核苷酸序列或至少部分核苷酸序列的克隆，可以通过合适表达系统在外源宿主中表达生产相应酶或者提高生物活性化合物产量。这些外源宿主包括大肠杆菌、链霉菌、假单胞菌、芽孢杆菌、酵母及动植物等。A clone comprising the nucleotide sequence provided by the present invention or at least part of the nucleotide sequence can be expressed in a foreign host by a suitable expression system to produce the corresponding enzyme or increase the yield of biologically active compounds. These foreign hosts include Escherichia coli, Streptomyces, Pseudomonas, Bacillus, yeast, animals and plants.

本发明提供的氨基酸序列可以用于分离所需蛋白并可用于抗体制备。The amino acid sequences provided by the present invention can be used to isolate the desired protein and can be used for antibody preparation.

包含本发明所提供的氨基酸序列或者至少部分序列的多肽，可能在去除或者替代某些氨基酸之后仍有生物活性甚至新的生物活性，或提高了产量或优化了蛋白动力学特征或其他致力于得到的性质。The polypeptide comprising the amino acid sequence provided by the present invention or at least part of the sequence may still have biological activity or even new biological activity after removing or replacing certain amino acids, or improve the yield or optimize the protein kinetic characteristics or other efforts to obtain nature.

包含本发明所提供的核苷酸序列或部分核苷酸序列的基因或基因簇可以在异源宿主中表达及揭示它们在宿主代谢中的功能。Genes or gene clusters comprising nucleotide sequences or partial nucleotide sequences provided by the present invention can be expressed in heterologous hosts and their function in host metabolism can be revealed.

包含本发明所提供核苷酸序列或至少部分核苷酸序列的基因或基因簇可以通过DNA重组技术构建重组载体，以获得新型生物合成途径，也可以通过插入、置换、缺失或失活，进而获得其他基于生物合成途径的新结构化合物。The gene or gene cluster comprising the nucleotide sequence or at least part of the nucleotide sequence provided by the present invention can construct a recombinant vector by DNA recombination technology to obtain a new biosynthetic pathway, or can be inserted, replaced, deleted or inactivated, and then Obtain other new structural compounds based on biosynthetic pathways.

本发明的第二个目的是提供非典型角环素类化合物nenestatin A的生物合成基因簇在制备非典型角环素类化合物nenestatin A或其类似物中的应用。The second object of the present invention is to provide the application of the biosynthetic gene cluster of the atypical keratin compound nenestatin A in the preparation of the atypical keratin compound nenestatin A or its analogs.

本发明的第三个目的是提供一种蒽醌氧化酶基因nes27，其特征在于，其核苷酸序列如SEQ ID NO.1的第32561-33304位的碱基序列所示。The third object of the present invention is to provide an anthraquinone oxidase gene nes27, which is characterized in that its nucleotide sequence is shown in the base sequence of positions 32561-33304 of SEQ ID NO.1.

本发明的第四个目的是提供上述蒽醌氧化酶基因nes27编码的蒽醌氧化酶NES27。The fourth object of the present invention is to provide anthraquinone oxidase NES27 encoded by the above-mentioned anthraquinone oxidase gene nes27.

本发明的第五个目的是提供蒽醌氧化酶NES27在催化式1所示的化合物ethyl-dehydrorabelomycin形成式2所示的化合物ethyl-kinobscurinone中的应用The fifth object of the present invention is to provide the application of anthraquinone oxidase NES27 in catalyzing the compound ethyl-dehydrorabelomycin shown informula 1 to form the compound ethyl-kinobscurinone shown informula 2

本发明的第六个目的是提供一种缺失上述蒽醌氧化酶基因nes27的基因工程菌。The sixth object of the present invention is to provide a genetically engineered bacterium lacking the above-mentioned anthraquinone oxidase gene nes27.

所述的基因工程菌优选为小单孢菌(Micromonospora echinospora)SCSIO04089。The genetically engineered bacteria are preferably Micromonospora echinospora SCSIO04089.

本发明的第七个目的是提供缺失上述的蒽醌氧化酶基因nes27的基因工程菌在制备式1所示的化合物ethyl-dehydrorabelomycin中的应用The seventh object of the present invention is to provide the application of genetically engineered bacteria lacking the above-mentioned anthraquinone oxidase gene nes27 in the preparation of the compound ethyl-dehydrorabelomycin shown informula 1

总之本发明所提供的包括nenestatin A生物合成相关的所有基因和蛋白信息，可以帮助人们理解角环素类天然产物的生物合成机制，为进一步遗传改造提供材料和理论基础。本发明所提供的基因和蛋白质可以用来寻找和发现可用于医药、卫生或农业的nenestatin A类化合物、基因或蛋白。In conclusion, the information provided by the present invention, including all genes and proteins related to the biosynthesis of nenestatin A, can help people understand the biosynthesis mechanism of keratin-like natural products, and provide materials and theoretical basis for further genetic modification. The genes and proteins provided by the present invention can be used to find and discover nenestatin A class compounds, genes or proteins that can be used in medicine, health or agriculture.

本发明的小单孢菌(Micromonospora echinospora)SCSIO 04089于2017年1月18日保藏于广东省微生物菌种保藏中心(GDMCC)，地址：广州市先烈中路100号大院59号楼5楼，其保藏编号为GDMCC No：60142。The Micromonospora echinospora SCSIO 04089 of the present invention was deposited in the Guangdong Provincial Microbial Culture Collection Center (GDMCC) on January 18, 2017, and the address is: 5th Floor, Building 59, No. 100, Xianlie Middle Road, Guangzhou City. The deposit number is GDMCC No: 60142.

附图说明：Description of drawings:

图1是nenestatin A和ethyl-dehydrorabelomycin的化学结构式。Figure 1 shows the chemical structures of nenestatin A and ethyl-dehydrorabelomycin.

图2是nenestatin A产生菌株小单孢菌(M.echinospora)SCSIO 04089最适发酵培养基的摸索；培养基N4为最适培养基，其中1表示化合物nenestatin A。Figure 2 is the exploration of the optimum fermentation medium for the nenestatin A producing strain M. echinospora SCSIO 04089; medium N4 is the optimum medium, wherein 1 represents the compound nenestatin A.

图3是nenestatin A生物合成基因簇示意图。包括了重叠覆盖nenestatin A生物合成基因簇的阳性克隆子pCSG4102、pCSG4103和pCSG4104。其中，nes1、nes5、nes9～11、nes19、nes30、nes39和nes59为function unknown，nes2～4、nes14、nes35～38为transportand resistance，nes6、nes7和nes58为regulation，nes8、nes13、nes41～48、nes50、nes51和nes60为sugar biosynthesis，nes12、nes15～17和nes20～27为tailoring ofpolyketide scaffold，nes18为dimerization，nes28、nes29、nes31～34为diazo groupbiosynthesis，nes40和nes49为glycosyltransferases，nes52～57为type II polyketidesynthase and starter unit biosunthesis，orf(-1)和orf(+1)为unrelated genes。Figure 3 is a schematic diagram of the nenestatin A biosynthetic gene cluster. Positive clones pCSG4102, pCSG4103 and pCSG4104 overlapping the nenestatin A biosynthetic gene cluster were included. Among them, nes1, nes5, nes9～11, nes19, nes30, nes39 and nes59 are function unknown, nes2～4, nes14, nes35～38 are transport and resistance, nes6, nes7 and nes58 are regulation, nes8, nes13, nes41～48, nes50, nes51 and nes60 are sugar biosynthesis, nes12, nes15-17 and nes20-27 are tailoring of polyketide scaffold, nes18 is dimerization, nes28, nes29, nes31-34 are diazo group biosynthesis, nes40 and nes49 are glycosyltransferases, and nes52-57 are type II polyketidesynthase and starter unit biosunthesis, orf(-1) and orf(+1) are unrelated genes.

图4是nenestatin A生物合成基因簇的鉴定。(A)nes52-54基因缺失示意图；(B)nes52-54基因缺失PCR验证图，泳道M为DL2000DNA Marker，泳道1模板为野生菌株M.echinospora SCSIO 04089，泳道2模板为nes52-54基因缺失突变株NES001；(C)HPLC检测野生菌株M.echinospora SCSIO 04089((i)Wild strain)和突变株NES001((ii)Δnes52-54)，其中1表示化合物nenestatin A。Figure 4 is the identification of nenestatin A biosynthetic gene clusters. (A) Schematic diagram of nes52-54 gene deletion; (B) nes52-54 gene deletion PCR verification diagram, lane M is DL2000 DNA Marker,lane 1 template is wild strain M.echinospora SCSIO 04089,lane 2 template is nes52-54 gene deletion mutation Strain NES001; (C) HPLC detection of wild strain M. echinospora SCSIO 04089 ((i) Wild strain) and mutant strain NES001 ((ii) Δnes52-54), wherein 1 represents the compound nenestatin A.

图5是蒽醌氧化酶nes27基因缺失突变株的鉴定。(A)nes27基因缺失示意图；(B)nes27基因缺失PCR验证图，泳道M为DL2000DNA Marker，泳道1模板为突变株NES002，泳道2模板为野生菌株M.echinospora SCSIO 04089；(C)HPLC检测野生菌株M.echinosporaSCSIO 04089((i)Wild strain)和突变株NES002((ii)Δnes27)，其中1表示化合物nenestatin A，2表示化合物ethyl-dehydrorabelomycin。Figure 5 is the identification of anthraquinone oxidase nes27 gene deletion mutants. (A) Schematic diagram of nes27 gene deletion; (B) nes27 gene deletion PCR verification diagram, lane M is DL2000 DNA Marker,lane 1 template is mutant NES002,lane 2 template is wild strain M.echinospora SCSIO 04089; (C) HPLC detected wild Strain M. echinosporaSCSIO 04089 ((i) Wild strain) and mutant strain NES002 ((ii) Δnes27), wherein 1 represents the compound nenestatin A and 2 represents the compound ethyl-dehydrorabelomycin.

图6是推测的nenestatin A的生物合成途径。Figure 6 is a putative biosynthetic pathway of nenestatin A.

图7是nenestatin A质谱图。Figure 7 is a mass spectrum of nenestatin A.

图8是nenestatin A在DMSO-d₆中NMR谱图。A为nenestatin A的H谱；B为nenestatinA的C谱；C为nenestatin A的DEPT 135谱；D为nenestatin A的HSQC谱；E为nenestatin A的COSY谱；F为nenestatin A的HMBC谱。Figure 8 is a NMR spectrum of nenestatin A in DMSO-d₆ . A is the H spectrum of nenestatin A; B is the C spectrum of nenestatin A; C is theDEPT 135 spectrum of nenestatin A; D is the HSQC spectrum of nenestatin A; E is the COSY spectrum of nenestatin A; F is the HMBC spectrum of nenestatin A.

图9是nenestatin A在CD₃OD中NMR谱图。A为nenestatin A的H谱；B为nenestatinA的C谱；C为nenestatin A的DEPT 135谱；D为nenestatin A的HSQC谱；E为nenestatin A的COSY谱；F为nenestatin A的HMBC谱。Figure 9 is a NMR spectrum of nenestatin A in_CD3OD . A is the H spectrum of nenestatin A; B is the C spectrum of nenestatin A; C is theDEPT 135 spectrum of nenestatin A; D is the HSQC spectrum of nenestatin A; E is the COSY spectrum of nenestatin A; F is the HMBC spectrum of nenestatin A.

图10是ethyl’-dehydrorabelomycin质谱图。Figure 10 is a mass spectrum of ethyl'-dehydrorabelomycin.

图11是ethyl’-dehydrorabelomycin NMR谱图。A为ethyl’-dehydrorabelomycin的H谱；B为ethyl’-dehydrorabelomycin的C谱；C为ethyl’-dehydrorabelomycin的DEPT135谱；D为ethyl’-dehydrorabelomycin的HSQC谱；E为ethyl’-dehydrorabelomycin的COSY谱；F为ethyl’-dehydrorabelomycin的HMBC谱。Figure 11 is an ethyl'-dehydrorabelomycin NMR spectrum. A is the H spectrum of ethyl'-dehydrorabelomycin; B is the C spectrum of ethyl'-dehydrorabelomycin; C is the DEPT135 spectrum of ethyl'-dehydrorabelomycin; D is the HSQC spectrum of ethyl'-dehydrorabelomycin; E is the COSY spectrum of ethyl'-dehydrorabelomycin; F is the HMBC spectrum of ethyl'-dehydrorabelomycin.

具体实施方式：Detailed ways:

以下实施例是对本发明的进一步说明，而不是对本发明保护范围的限制。The following examples are to further illustrate the present invention, rather than limit the protection scope of the present invention.

1.Nenestatin A产生菌株M.echinospora SCSIO 04089最适发酵培养基优化1. Optimization of optimal fermentation medium for Nenestatin A-producing strain M.echinospora SCSIO 04089

菌株M.echinospora SCSIO 04089分离自中国南海北部深海沉积物中，为充分挖掘该菌株次级代谢产物产生能力，通过8种培养基筛选最适培养基，培养基组成分别为：(1)N4培养基：淀粉15g，鱼蛋白胨8g，细菌学蛋白胨5g，甘油6mL，CaCO₃ 2g，KBr 0.2g，海盐10g，加水定容至1L，pH7.0；(2)Am6-1培养基：可溶性淀粉20g，甘油10g，甘氨酸0.1g，丙氨酸0.1g，苏氨酸0.1g；异亮氨酸0.1g，酵母粉5g，CaCO₃ 5g，海盐30g，加水定容至1L，pH7.0；(3)Am6-4培养基：甘油10g，蛋白胨5g，甘氨酸0.1g，丙氨酸0.1g，CaCO₃ 5g，海盐30g，加水定容至1L，pH7.0；(4)RA培养基：葡萄糖10g，可溶性淀粉20g，麦芽提取粉10g，玉米粉5g，海盐30g，微量元素(每升微量元素含有FeSO₄·H₂O 0.1g，MnCl₂·4H₂O 0.1g和ZnSO₄·7H₂O0.1g，余量为水)100μL，加水定容至1L，pH 7.2-7.4；(5)ISP2培养基：葡萄糖4g，麦芽提取粉10g，酵母粉5g，海盐30g，加水定容至1L，pH7.2-7.4；(6)ISP4培养基：淀粉10g，(NH₄)₂SO₄2g，K₂HPO₄ 1g，CaCO₃ 1g，MgSO₄·7H₂O 1g，微量元素(每升微量元素含有FeSO₄·H₂O 0.1g，MnCl₂·4H₂O 0.1g，ZnSO₄·7H₂O 0.1g，余量为水)100μL，海盐30g，加水定容至1L，pH 7.2-7.4；(7)YMS培养基：酵母粉4g，麦芽提取物10g，可溶性淀粉5g、氯化钴50mg，海盐30g，加水定容至1L，pH 7.2-7.4；(8)AM2ab培养基：可溶性淀粉5g、葡萄糖20g、酵母粉2g、蛋白胨2g、黄豆粉5g、MgSO₄·7H₂O0.5g、K₂HPO₄ 0.5g、氯化钠4g、海盐30g、碳酸钙2g，加水定容至1L，pH7.2-7.4。28℃，200r/min振荡培养7天的发酵粗提物HPLC检测显示N4培养基有明显的次级代谢产物产生(图2)。The strain M.echinospora SCSIO 04089 was isolated from the deep-sea sediments in the northern South China Sea. In order to fully exploit the secondary metabolite production capacity of the strain, the optimal medium was screened through 8 kinds of medium. The medium composition was: (1) N4 culture Base: starch 15g, fish peptone 8g, bacteriological peptone 5g, glycerol 6mL, CaCO₃ 2g, KBr 0.2g, sea salt 10g, add water to 1L, pH 7.0; (2) Am6-1 medium: soluble starch 20g , glycerol 10g, glycine 0.1g, alanine 0.1g, threonine 0.1g; isoleucine 0.1g, yeast powder 5g, CaCO₃ 5g, sea salt 30g, add water to make up to 1L, pH7.0; (3 ) Am6-4 medium: glycerol 10g, peptone 5g, glycine 0.1g, alanine 0.1g, CaCO₃ 5g, sea salt 30g, add water to make up to 1L, pH 7.0; (4) RA medium: glucose 10g, Soluble starch 20g, malt extract powder 10g, corn flour 5g, sea salt 30g, trace elements (each liter of trace elements contains FeSO₄ ·H₂ O 0.1g, MnCl₂ ·4H₂ O 0.1g and ZnSO₄ ·7H₂ O 0.1g (5) ISP2 medium: glucose 4g, malt extract powder 10g, yeast powder 5g, sea salt 30g, add water to volume to 1L, pH7.2 -7.4; (6) ISP4 medium: starch 10g, (NH₄ )₂ SO₄ 2g, K₂ HPO₄ 1g, CaCO₃ 1g, MgSO₄ ·7H₂ O 1g, trace elements (each liter of trace elements contains FeSO_{4 )} ·H₂ O 0.1g, MnCl₂ ·4H₂ O 0.1g, ZnSO₄ ·7H₂ O 0.1g, the balance is water) 100μL, sea salt 30g, add water to make up to 1L, pH 7.2-7.4; (7) YMS Medium: yeast powder 4g, malt extract 10g, soluble starch 5g, cobalt chloride 50mg, sea salt 30g, add water to make up to 1L, pH 7.2-7.4; (8) AM2ab medium: soluble starch 5g, glucose 20g, yeast Powder 2g, peptone 2g, soybean powder 5g, MgSO₄ ·7H₂ O 0.5g, K₂ HPO₄ 0.5g, sodium chloride 4g, sea salt 30g, calcium carbonate 2g, add water to make up to 1L, pH7.2-7.4. The HPLC detection of the fermentation crude extract after 7 days of shaking culture at 28°C and 200r/min showed that the N4 medium had obvious secondary metabolite production (Fig. 2).

2.化合物Nenestatin A的分离鉴定2. Isolation and identification of the compound Nenestatin A

将菌种M.echinospora SCSIO 04089接种至ISP4平板培养基上，28℃培养7天，接种于一级种子培养基(A1培养基：淀粉10克，酵母提取粉4克，细菌学蛋白胨2克，海盐30克，加水定容至1L，pH7.0)中(50mL培养基/250mL三角瓶)，28℃，200r/min振荡培养3天作为一级发酵种子，吸取该一级种子液2mL接种到二级种子培养基(A1培养基)中(50mL培养基/250mL三角瓶)，28℃，200r/min振荡培养2天作为二级发酵种子，将生长良好的二级种子液以20％(v/v)的接种量接入发酵培养基(N4培养基：淀粉15克，鱼蛋白胨8克，细菌学蛋白胨5克，甘油6毫升，碳酸钙2克，溴化钾0.2克，海盐10克，加水定容至1升，pH7.0)中(200mL培养基/1000mL三角瓶)，28℃，200r/min培养7～10天。The strain M.echinospora SCSIO 04089 was inoculated on the ISP4 plate medium, cultivated at 28°C for 7 days, and inoculated in the first-class seed medium (A1 medium: 10 grams of starch, 4 grams of yeast extract powder, 2 grams of bacteriological peptone, 30 grams of sea salt, add water and dilute to 1L, pH 7.0) (50mL medium/250mL conical flask), 28 ℃, 200r/min shaking culture for 3 days as the first-level fermentation seeds, suck 2mL of the first-level seed liquid and inoculate it into In the secondary seed medium (A1 medium) (50mL medium/250mL conical flask), 28 ℃, 200r/min shaking culture for 2 days as the secondary fermentation seeds, the well-grown secondary seed liquid was 20% (v /v) of the inoculum into the fermentation medium (N4 medium: starch 15 g, fish peptone 8 g, bacteriological peptone 5 g,glycerol 6 ml, calcium carbonate 2 g, potassium bromide 0.2 g, sea salt 10 g, Add water to dilute to 1 liter, pH 7.0) (200 mL medium/1000 mL conical flask), and culture at 28° C., 200 r/min for 7 to 10 days.

发酵结束，离心收集发酵液，上清液过大孔树脂吸附3次，后用丙酮洗脱3次，旋转蒸发回收丙酮，水相用丁酮萃取，旋转蒸发丁酮萃取液得到发酵液浸膏；菌丝体用丙酮浸泡，旋转蒸发回收丙酮，水相用丁酮萃取，旋转蒸发丁酮萃取液得到菌丝体浸膏，合并得粗浸膏。粗浸膏首先使用正向硅胶(100-200目，100g)梯度洗脱(CHCl₃/MeOH/H₂O，95/5/0.5，90/10/1，V/V，每次600mL)得到2个组分(Fr.1-Fr2)，其中组分Fr2经过ODS column(50μm,YMC*GEL ODS-A-HG,30g)在Agela Cheetahtm MP200MPLC工作站上进行洗脱(CH₃OH/H₂O，体积分数为10％-100％)进行1.5小时的梯度洗脱，获得5个亚馏分(Fr.2.1to Fr.2.5)，其中组分Fr2.2经过反向高压制备得到nenestatin A[制备条件：Phenomenex ODS column(250mm×10.0mm i.d.,5μm；Phenomenex Phenomenex ODS column(250mm×10.0mm i.d.,5μm；Phenomenex，流动相A,0.1％formic acid in water；流动相B,90％CH₃CN in water；在流动相B为55％的条件下等度洗脱；流速为2.5mL/min，保留时间为13min时得到nenestatinA)。After fermentation, the fermentation broth was collected by centrifugation. The supernatant was adsorbed by the macroporous resin for three times, and then eluted with acetone for three times. The acetone was recovered by rotary evaporation. The mycelium is soaked in acetone, the acetone is recovered by rotary evaporation, the aqueous phase is extracted with butanone, and the butanone extract is obtained by rotary evaporation to obtain a mycelium extract, which is combined to obtain a crude extract. The crude extract was first obtained by gradient elution (CHCl₃ /MeOH/H₂ O, 95/5/0.5, 90/10/1, V/V, 600 mL each time) using normal silica gel (100-200 mesh, 100 g) 2 components (Fr.1-Fr2), wherein the component Fr2 was eluted (CH₃ OH/H₂ O) on an Agela Cheetahtm MP200MPLC workstation through an ODS column (50μm, YMC*GEL ODS-A-HG, 30g). , the volume fraction is 10%-100%) for 1.5 hours of gradient elution to obtain 5 sub-fractions (Fr.2.1 to Fr.2.5), in which the fraction Fr2.2 was prepared by reverse high pressure to obtain nenestatin A [preparation conditions : Phenomenex ODS column (250mm×10.0mm id, 5μm; Phenomenex Phenomenex ODS column (250mm×10.0mm id, 5μm; Phenomenex, mobile phase A, 0.1% formic acid in water; mobile phase B, 90% CH₃ CN in water ; Isocratic elution under the condition that the mobile phase B is 55%; the flow rate is 2.5mL/min, and the retention time is 13min to obtain nenestatinA).

3.Nenestatin A的生物合成基因簇的克隆与功能预测3. Cloning and functional prediction of the biosynthetic gene cluster of Nenestatin A

Nenestatin A为非典型角环素类化合物，通过对菌株M.echinospora SCSIO04089进行全基因组扫描和注释，分析到65kb的Nenestatin A生物合成基因簇，共包含60个开放阅读框(open reading frames，ORFs)，详细基因注释如表1所示。Nenestatin A is an atypical keratin-like compound. Through genome-wide scanning and annotation of the strain M.echinospora SCSIO04089, a 65kb Nenestatin A biosynthetic gene cluster was analyzed, containing a total of 60 open reading frames (ORFs). , and the detailed gene annotations are shown in Table 1.

表1Nenestatin A生物合成基因簇中基因功能预测Table 1 Gene function prediction in Nenestatin A biosynthetic gene cluster

依据分析到的基因簇序列设计特异引物(表2)，从构建的菌株M.echinosporaSCSIO 04089基因组文库中筛选出包含有目的片段的阳性克隆，阳性克隆子经末端测序及限制性内切酶酶切分析，最终确定3个cosmids，即pCSG4102，pCSG4103，pCSG4104可重叠覆盖nenestatin A的生物合成基因簇(图3)。Specific primers (Table 2) were designed according to the analyzed gene cluster sequences, and positive clones containing the target fragments were screened from the constructed strain M.echinosporaSCSIO 04089 genomic library. The positive clones were sequenced and cut with restriction enzymes. The analysis finally determined that three cosmids, namely pCSG4102, pCSG4103, and pCSG4104, could overlap and cover the biosynthetic gene cluster of nenestatin A (Fig. 3).

表2本发明中使用的引物Table 2 Primers used in the present invention

4.Nenestatin A的生物合成基因簇的确定4. Determination of the biosynthetic gene cluster of Nenestatin A

为进一步确定所克隆的nenestatin A生物合成基因簇的正确性，构建了菌株M.echinospora SCSIO 04089的遗传操作系统，并选取其中的聚酮链延伸基因nes52-54进行缺失，构建突变株NES001，突变过程示意图和突变株PCR验证如图4(A)和4(B)所示，HPLC显示突变株NES001丧失了产生nenestatin A的能力(图4(C)，(ii))，实验结果进一步证明克隆到的nenestatin A的生物合成基因簇的正确性。In order to further confirm the correctness of the cloned nenestatin A biosynthetic gene cluster, the genetic operating system of the strain M.echinospora SCSIO 04089 was constructed, and the polyketide chain extension gene nes52-54 was selected for deletion, and the mutant strain NES001 was constructed. The schematic diagram of the process and the PCR verification of the mutant strain are shown in Figures 4(A) and 4(B), HPLC showed that the mutant strain NES001 lost the ability to produce nenestatin A (Figure 4(C), (ii)), and the experimental results further proved that the clone To the correctness of the biosynthetic gene cluster of nenestatin A.

5.基因nes27功能验证及代谢中间体分离鉴定5. Functional verification of gene nes27 and isolation and identification of metabolic intermediates

选取生物合成基因簇中的蒽醌氧化酶基因nes27进行中断缺失，构建双交换突变株，突变过程示意图和突变株PCR验证如图5(A)和5(B)所示。构建的Δnes27突变株NES002丧失了产生nenestatin A的能力(图5(C)，(ii))，而累积一个新的代谢中间产物，经发酵分离纯化，质谱、NMR解析确定了单体化合物的结构式。命名为ethyl-dehydrorabelomycin(图1)，基因nes27的缺失结果进一步证明克隆到的nenestatin A的生物合成基因簇的正确性。The anthraquinone oxidase gene nes27 in the biosynthetic gene cluster was selected for interruption and deletion, and a double-crossover mutant was constructed. The schematic diagram of the mutation process and the PCR verification of the mutant are shown in Figures 5(A) and 5(B). The constructed Δnes27 mutant strain NES002 lost the ability to produce nenestatin A (Fig. 5(C), (ii)), but accumulated a new metabolic intermediate, which was purified by fermentation, and the structural formula of the monomer compound was determined by mass spectrometry and NMR analysis. . Named ethyl-dehydrorabelomycin (Figure 1), the deletion of the gene nes27 further confirmed the correctness of the cloned nenestatin A biosynthetic gene cluster.

6.Nenestatin A的生物合成机制推导6. Deduction of the biosynthetic mechanism of Nenestatin A

根据生物信息学分析，推导了nenestatin A的生物合成机制。聚酮链的起始单元为丙酰-CoA，由甲基丙二酰CoA通过Nes55-Nes57脱羧生成，随后通过Nes52-Nes54添加9个延伸单元(丙二酰CoA)生成全长聚酮链，在经过酮基还原(Nes21)、环化作用(Nes20和Nes24)和氧化作用生成ethyl-dehydrorabelomycin，经过Nes27氧化开环、闭环生成ethyl-kinobscurinone，在重氮形成酶Nes28、Nes29和Nes31-Nes34等6个酶的作用下添加重氮基团生成化合物ethyl-prekinamycin，随后经过羟化，环氧，环氧打开，脱去重氮基团和糖基化等生物合成步骤，生成终产物nenestatin A，同时推导了稀有糖基4-O-甲基-angolosamine的合成途径。推导的合成途径如图6所示。Based on bioinformatics analysis, the biosynthetic mechanism of nenestatin A was deduced. The starting unit of the polyketone chain is propionyl-CoA, which is generated by decarboxylation of methylmalonyl CoA through Nes55-Nes57, and then 9 extension units (malonyl-CoA) are added through Nes52-Nes54 to generate a full-length polyketone chain, After ketone reduction (Nes21), cyclization (Nes20 and Nes24) and oxidation to generate ethyl-dehydrorabelomycin, after Nes27 oxidative ring opening and ring closure to generate ethyl-kinobscurinone, in the diazo-forming enzymes Nes28, Nes29 and Nes31-Nes34, etc. Under the action of 6 enzymes, the diazo group is added to generate the compound ethyl-prekinamycin, and then through the biosynthesis steps such as hydroxylation, epoxy, epoxy opening, removal of the diazo group and glycosylation, the final product nenestatin A is generated, At the same time, the synthetic route of rare glycosyl 4-O-methyl-angolosamine was deduced. The deduced synthetic route is shown in Figure 6.

以下进一步提供实施实例，这些实施实例有助于理解本发明，仅用作说明而不限制本发明的应用范围。Embodiments are further provided below, which are helpful for understanding the present invention, are only used for illustration and do not limit the scope of application of the present invention.

实施例1：Nenestatin A生产菌M.echinospora SCSIO 04089基因组DNA的提取Example 1: Extraction of genomic DNA of Nenestatin A producing strain M.echinospora SCSIO 04089

将新鲜M.echinospora SCSIO 04089的菌丝体按照5％的接种量接种于50mL的TSB培养基(配方：大豆蛋白胨5g，胰蛋白胨15g，蔗糖100g，葡萄糖2.5g，K₂HPO₄ 2.5g，海盐30g，加水定容至1L，pH 7.0)中，28℃～30℃，振荡培养3-4d，4000rpm离心10min收集菌丝体。菌丝体用STE溶液(75mM NaCl，25mM EDTA，20mM Tris-Cl，余量为水，pH＝8.0)洗涤两次，向洗涤后的菌丝体中加入30mL STE溶液和终浓度3mg/mL的溶菌酶，涡旋均匀，37℃温浴3h，加入至终浓度0.1-0.2mg/mL的蛋白酶K，混匀，37℃温浴10min，加入至终浓度1％-2％的SDS，混匀，放入55℃水浴约1h，期间颠倒数次。加入等体积酚-氯仿-异戊醇(V/V/V＝25:24:1)，混合均匀，置于冰上冷却30min。12000rpm，4℃离心10min，用剪过的大口径枪头小心吸取上清到新的离心管中，用同样的方法反复处理3次，用等体积的氯仿洗涤两次，12000rpm，4℃离心10min。用剪过的大口径枪头将水相吸出转移至新的离心管，加入1/10体积3mol/L NaAc(pH＝5.2)，混匀后再加入等体积的异丙醇，混匀后冰上放置，沉淀DNA。小心地用玻璃棒将DNA纤维团转移至新的离心管中，用70％乙醇洗涤两次，将液体倾出，在37℃下略微烘干，加5mL TE(10mM Tris-HCl，1mM EDTA，余量为水，pH 8.0)溶解，并加入3-5U的RNA酶，由此得到M.echinospora SCSIO 04089基因组DNA。The mycelium of fresh M.echinospora SCSIO 04089 was inoculated in 50 mL of TSB medium (formula: soy peptone 5 g, tryptone 15 g, sucrose 100 g, glucose 2.5 g, K₂ HPO₄ 2.5 g, sea salt 30g, add water to dilute to 1L, pH 7.0), 28℃～30℃, shake culture for 3-4d, centrifuge at 4000rpm for 10min to collect mycelium. Mycelium was washed twice with STE solution (75mM NaCl, 25mM EDTA, 20mM Tris-Cl, the balance was water, pH=8.0), and 30mL STE solution and 3mg/mL final concentration of 30mL STE solution were added to the washed mycelia. Lysozyme, vortex evenly, incubate at 37 °C for 3 h, add proteinase K to a final concentration of 0.1-0.2 mg/mL, mix well, incubate at 37 °C for 10 min, add SDS to a final concentration of 1%-2%, mix well, put Into the 55 ℃ water bath for about 1h, inversion several times during the period. An equal volume of phenol-chloroform-isoamyl alcohol (V/V/V=25:24:1) was added, mixed uniformly, and cooled on ice for 30 min. Centrifuge at 12,000 rpm for 10 min at 4°C, carefully pipette the supernatant into a new centrifuge tube with a large-diameter pipette tip that has been cut, repeat the treatment three times with the same method, wash twice with an equal volume of chloroform, and centrifuge at 12,000 rpm for 10 min at 4°C. . Use a cut large-diameter pipette tip to suck out the aqueous phase and transfer it to a new centrifuge tube, add 1/10 volume of 3mol/L NaAc (pH=5.2), and then add an equal volume of isopropanol after mixing. Place on top to precipitate DNA. Carefully transfer the DNA fiber mass to a new centrifuge tube with a glass rod, wash twice with 70% ethanol, pour out the liquid, dry it slightly at 37 °C, add 5 mL of TE (10 mM Tris-HCl, 1 mM EDTA, The balance is water, pH 8.0) is dissolved, and 3-5U of RNase is added, thereby obtaining M. echinospora SCSIO 04089 genomic DNA.

实施例2：Nenestatin A生产菌M.echinospora SCSIO 04089全基因组文库的构建Example 2: Construction of the whole genome library of Nenestatin A producing strain M.echinospora SCSIO 04089

首先通过一系列的稀释实验确定限制性核酸内切酶Sau3AI的用量，在20μL体系中，含有17μL的基因组DNA，2μL的10×反应缓冲液和1μL不同稀释度的Sau3A I，其终止反应为4μL 0.5mol/L EDTA和合适的上样缓冲液。通过摸索确定了0.025-0.05U的酶活单位比较合适。在此基础上通过大量部分酶切得到略大于40kb的基因组DNA片段，用去磷酸化酶进行去磷酸化处理。First, the amount of restriction endonuclease Sau3AI was determined by a series of dilution experiments. In a 20 μL system, containing 17 μL of genomic DNA, 2 μL of 10× reaction buffer and 1 μL of Sau3A I at different dilutions, the termination reaction was 4 μL 0.5mol/L EDTA and appropriate loading buffer. Through exploration, it is determined that the enzyme activity unit of 0.025-0.05U is more suitable. On this basis, a genomic DNA fragment slightly larger than 40kb was obtained by a large number of partial enzyme digestion, and dephosphorylated with dephosphorylase.

用于构建文库的载体SuperCos I质粒先用限制性核酸内切酶XbaI从两个cos序列中间切开，然后进行去磷酸化处理，再从多克隆位点处用限制性核酸内切酶BamHI切开，获得两个臂。处理后的载体与之前制备的部分酶切的约40kb的基因组DNA片段连接过夜，连接体系为10μL，含有1.25μg制备的基因组DNA和0.5μg处理后的SuperCos I质粒，1μL的10×Buffer，0.3U的连接酶。连接产物在65℃处理15min，使其失活。从-80℃冰箱中取出一管包装混合物(50μL)置于冰上，将包装混合物在指间迅速融化，小心吸取一半包装混合物(25μL)至一个新的离心管中，加入10μL热处理后的连接产物，其余包装混合物于-80℃保存。小心混匀，30℃温浴90min，加入另外一半包装混合物(25μL)，30℃温浴90min。加入500μL噬菌体稀释缓冲液(100mmol/L NaCl，10mmol/L MgCl₂，10mmol/L Tris-HCl，余量为水，pH＝8.3)，再加入25μL氯仿，轻轻混匀，于4℃保存。The vector SuperCos I plasmid used to construct the library was first cut from the middle of the two cos sequences with the restriction endonuclease XbaI, then dephosphorylated, and then cut with the restriction endonuclease BamHI from the multiple cloning site Open to get two arms. The treated vector was ligated with the previously prepared partially digested genomic DNA fragment of about 40 kb overnight. The ligation system was 10 μL, containing 1.25 μg of the prepared genomic DNA and 0.5 μg of the treated SuperCos I plasmid, 1 μL of 10×Buffer, 0.3 U ligase. The ligation product was inactivated by treatment at 65°C for 15 min. Remove one tube of packing mix (50 μL) from the -80°C freezer and place on ice, thaw the packing mix quickly between your fingers, carefully pipette half of the packing mix (25 μL) into a new centrifuge tube, and add 10 μL of heat-treated ligation product, the rest of the packaging mix was stored at -80°C. Mix carefully, incubate at 30°C for 90 minutes, add the other half of the packaging mixture (25 μL), and incubate at 30°C for 90 minutes. Add 500 μL of phage dilution buffer (100 mmol/L NaCl, 10 mmol/L MgCl₂ , 10 mmol/L Tris-HCl, the balance is water, pH=8.3), then add 25 μL of chloroform, mix gently, and store at 4°C.

将冻存于-80℃的菌株E.coli LE392MP(Stratagene)涂布在LB培养基上复苏。包装反应前一天，挑取单克隆接种于LB培养基中(添加0.2％麦芽糖和10mM MgSO₄)37℃振荡培养过夜，包装反应当天，取5mL过夜培养的菌液加入到50mL新鲜的LB培养基中(添加0.2％麦芽糖和10mM MgSO₄)，37℃，200rpm振荡至培养物OD600达到0.8-1时，4℃保存备用。取100μL如上处理的宿主菌液和100μL适度稀释的包装液轻轻混匀，于37℃温浴15min，涂布于含有100μg/mL氨苄青霉素和50μg/mL卡那霉素的LB平板上，37℃培养过夜。将长出来的单个克隆子，用无菌牙签点种于含100μg/mL氨苄青霉素和50μg/mL卡那霉素的LB的96孔板上，37℃培养过夜，加入终浓度为20％的甘油，混合均匀，置于-80℃保存，由此得到Micromonosporaechinospora SCSIO 04089全基因组文库。The strain E. coli LE392MP (Stratagene) cryopreserved at -80°C was spread on LB medium for recovery. One day before the packaging reaction, pick a single clone and inoculate it in LB medium (add 0.2% maltose and 10 mM MgSO₄ ), shake and culture at 37°C overnight, on the day of the packaging reaction, take 5 mL of the overnight culture and add it to 50 mL of fresh LB medium (add 0.2% maltose and 10 mM MgSO₄ ), shake at 200 rpm at 37° C. until the OD600 of the culture reaches 0.8-1, and store at 4° C. for later use. Take 100 μL of the above-treated host bacterial solution and 100 μL of the moderately diluted packaging solution, mix gently, incubate at 37 °C for 15 minutes, and spread on LB plates containing 100 μg/mL ampicillin and 50 μg/mL kanamycin, at 37 °C Incubate overnight. The grown single clones were seeded with sterile toothpicks on LB 96-well plates containing 100 μg/mL ampicillin and 50 μg/mL kanamycin, cultured at 37°C overnight, and added with a final concentration of 20% glycerol. , mixed well, and stored at -80°C, thereby obtaining the Micromonosporaechinospora SCSIO 04089 whole genome library.

实施例3：从菌株M.echinospora SCSIO 04089基因组文库中筛选覆盖nenestatinA生物合成基因簇的阳性克隆子Example 3: Screening of positive clones covering the nenestatinA biosynthetic gene cluster from the genomic library of strain M.echinospora SCSIO 04089

将菌株M.echinospora SCSIO 04089基因组DNA送中国科学院北京基因组研究所进行全基因组扫描和注释，根据扫描和注释的结果，通过生物信息学分析，初步确定nenestatin A生物合成基因簇位于contig15和contig61上，contigs之间的缺口通过PCR填充连接。设计特异性引物4089S1-F/R，4089S3-F/R，4089S5-F/R(表2)筛选M.echinosporaSCSIO 04089全基因组文库，获得的阳性克隆子通过限制性核酸内切酶酶切及末端测序确定3个克隆子pCSG4102，pCSG4103和pCSG4104可重叠覆盖nenestatin A生物合成基因簇，并经测序，其核苷酸序列如SEQ ID NO.1的第3153-68576位的碱基序列所示。(图3)。The genome DNA of strain M.echinospora SCSIO 04089 was sent to Beijing Institute of Genomics, Chinese Academy of Sciences for genome-wide scanning and annotation. According to the results of scanning and annotation, the bioinformatics analysis preliminarily determined that the nenestatin A biosynthetic gene cluster was located on contig15 and contig61. The gaps between the contigs were connected by PCR filling. Design specific primers 4089S1-F/R, 4089S3-F/R, 4089S5-F/R (Table 2) to screen the M.echinosporaSCSIO 04089 whole genome library, and the obtained positive clones were digested with restriction endonuclease and the ends Sequencing confirmed that the three clones pCSG4102, pCSG4103 and pCSG4104 could overlap and cover the nenestatin A biosynthetic gene cluster, and after sequencing, their nucleotide sequences are shown in the base sequence of positions 3153-68576 of SEQ ID NO.1. (image 3).

实施例4：Nenestatin A生产菌M.echinospora SCSIO 04089遗传操作系统的建立，以敲除蒽酮氧化酶基因nes27为例：Example 4: The establishment of the genetic operating system of the Nenestatin A producing strain M.echinospora SCSIO 04089, taking the knockout of the anthrone oxidase gene nes27 as an example:

利用PCR-targeting的方法获得体外敲除突变株，根据获得的nenestatin A生物合成基因簇序列，设计nes27基因的敲除引物nes27-TarF/R(表2)，随后将构建好的敲除质粒转入接合转移的供体菌中，筛选阳性突变株。具体步骤如下：(1)将cosmid质粒pCSG4103(其核苷酸序列如SEQ ID NO.1第25405-59361位碱基所示)转入大肠杆菌E.coli BW25113/pIJ790中获得E.coli BW25113/pIJ790/pCSG4103重组菌株，用10mmol/L的L-阿拉伯糖诱导λ/red重组系统表达，并将其制备为电转感受态细胞待用。(2)用内切酶EcoRI和HindIII酶切质粒pIJ773，回收约1.4kb含有接合转移原点和阿泊拉霉素抗性基因的DNA片段，以此作为PCR模板，用引物nes27-TarF/R(表2)通过PCR扩增出1.4kb的PCR产物；50μL的PCR反应体系：高保真DNA聚合酶3U，10×Buffer 5μL，dNTPs 0.5mmol/L，DMSO 2.5μL，引物各0.5μmol/L，DNA模板约1ng，加水补至50μL。PCR反应条件为：预变性95℃，5min；扩增循环为94℃变性30s，58℃退火30s，72℃延伸90s，32个循环；最后72℃延伸10min。将1.4kb的PCR产物回收纯化待用。(3)将回收后的1.4kb PCR产物电转入步骤(1)中制备的感受态细胞使其发生重组，涂布于LB筛选平板(含100μg/mL氨苄青霉素，50μg/mL卡那霉素，50μg/mL阿泊拉霉素)上，37℃过夜培养。从平板上挑出阳性单克隆，使用验证引物nes27-TestF/R(表2)验证，抽提经PCR验证的质粒，命名为pCSG4117，该质粒中的nes27基因的部分片段被接合转移原点和阿泊拉霉素抗性基因取代。(4)将构建好的重组突变质粒pCSG4117转化到E.coli ET12567/pUZ8002中，构建成E.coli ET12567/pUZ8002/pCSG4117重组菌株，作为接合转移的供体菌。The in vitro knockout mutant was obtained by PCR-targeting, and the knockout primer nes27-TarF/R of the nes27 gene was designed according to the obtained nenestatin A biosynthetic gene cluster sequence (Table 2), and then the constructed knockout plasmid was transfected into Into the donor bacteria transferred by conjugation, the positive mutants were screened. The specific steps are as follows: (1) The cosmid plasmid pCSG4103 (the nucleotide sequence of which is shown in the bases 25405-59361 of SEQ ID NO.1) is transferred into E. coli E.coli BW25113/pIJ790 to obtain E.coli BW25113/ The pIJ790/pCSG4103 recombinant strain was induced to express by the λ/red recombinant system with 10 mmol/L L-arabinose, and it was prepared as an electrocompetent cell for use. (2) The plasmid pIJ773 was digested with endonucleases EcoRI and HindIII, and a DNA fragment of about 1.4 kb containing the junction transfer origin and the apramycin resistance gene was recovered, which was used as a PCR template, and the primers nes27-TarF/R ( Table 2) PCR product of 1.4kb was amplified by PCR; 50μL PCR reaction system: high-fidelity DNA polymerase 3U, 10×Buffer 5μL, dNTPs 0.5mmol/L, DMSO 2.5μL, primers 0.5μmol/L each, DNA The template is about 1 ng, and water is added to make up to 50 μL. The PCR reaction conditions were: pre-denaturation at 95°C for 5 min; amplification cycle was 32 cycles of denaturation at 94°C for 30s, annealing at 58°C for 30s, and extension at 72°C for 90s; and a final extension at 72°C for 10 min. The 1.4kb PCR product was recovered and purified for use. (3) The recovered 1.4kb PCR product was electroporated into the competent cells prepared in step (1) to recombine, and then spread on LB screening plates (containing 100 μg/mL ampicillin, 50 μg/mL kanamycin) , 50 μg/mL apramycin), cultured overnight at 37°C. The positive single clones were picked from the plate and verified using the verification primer nes27-TestF/R (Table 2), and the PCR-verified plasmid was extracted and named pCSG4117. Part of the nes27 gene fragment in this plasmid was conjugated to the transfer origin and A Polamycin resistance gene substitution. (4) The constructed recombinant mutant plasmid pCSG4117 was transformed into E.coli ET12567/pUZ8002 to construct E.coli ET12567/pUZ8002/pCSG4117 recombinant strain as the donor strain for conjugation transfer.

菌株M.echinospora SCSIO 04089在ATCC172培养基(酵母粉4g，葡萄糖4g，麦芽提取粉5g，FeSO₄·7H₂O 1mg，MnCl₂ 1mg，ZnSO₄·7H₂O 1mg，海盐30g，加水定容至1L，pH 7.0)平板中划线培养7天，长出的孢子用无菌棉签收集于含3％海盐的TSB培养基中(大豆蛋白胨5g，胰蛋白胨15g，蔗糖100g，葡萄糖2.5g，K₂HPO₄ 2.5g，海盐30g，加水定容至1L，pH 7.0)，涡旋振荡，分散孢子。过滤分离菌丝体和孢子，孢子悬浮于5mL含3％海盐的TSB培养基中，50℃热激10min，然后于28℃萌发4h，作为接合转移的受体菌。供体菌E.coli ET12567/pUZ8002/pCSG4117在50mL含50μg/mL卡那霉素，25μg/mL氯霉素和50μg/mL阿泊拉霉素的LB液体培养基中于37℃振荡培养4h至OD600值约为0.6时，4000rpm离心10min收集菌体，用LB清洗菌体3遍，悬浮于300μL LB培养基中，作为接合转移的供体菌。取上述受体菌400μL和供体菌100μL混合均匀，涂布于含有90mM MgSO₄不含任何抗生素的ISP4固体培养基上，吹干后，于28℃培养18h，然后将平板取出，用含有抗生素的水覆盖平板，其终浓度为15μg/mL阿泊拉霉素和20μg/mL甲氧苄氨嘧啶，吹干后，置于28℃培养箱中，培养5d后观察。Strain M.echinospora SCSIO 04089 in ATCC172 medium (yeast powder 4g, glucose 4g, malt extract powder 5g, FeSO₄ 7H₂ O 1mg, MnCl₂ 1mg, ZnSO₄ 7H₂ O 1mg, sea salt 30g, add water to make up 1L, pH 7.0) plates were streaked for 7 days, and the grown spores were collected with sterile cotton swabs in TSB medium containing 3% sea salt (soy peptone 5 g, tryptone 15 g, sucrose 100 g, glucose 2.5 g, K₂ HPO₄ 2.5g, sea salt 30g, add water to dilute to 1L, pH 7.0), vortex to disperse the spores. The mycelium and spores were separated by filtration, and the spores were suspended in 5 mL of TSB medium containing 3% sea salt, heat-shocked at 50°C for 10 minutes, and then germinated at 28°C for 4 hours as recipient bacteria for conjugation transfer. The donor bacteria E.coli ET12567/pUZ8002/pCSG4117 were cultured with shaking at 37°C for 4h to When the OD600 value was about 0.6, the cells were collected by centrifugation at 4000 rpm for 10 min, washed three times with LB, and suspended in 300 μL of LB medium to serve as the donor bacteria for conjugation transfer. Take 400 μL of the above-mentioned recipient bacteria and 100 μL of the donor bacteria and mix them evenly, spread them on ISP4 solid medium containing 90 mM MgSO₄ without any antibiotics, dry them, and cultivate them at 28 °C for 18 h. The plate was covered with water with a final concentration of 15 μg/mL apramycin and 20 μg/mL trimethoprim. After drying, it was placed in a 28 °C incubator and observed after 5 days of incubation.

当接合转移平板上长出菌落后，用无菌牙签将其转接到含有35μg/mL阿泊拉霉素和50μg/mL甲氧苄氨嘧啶的ATCC172号培养基(淀粉20g，葡萄糖10g，酵母提取物5g，Aobox酪蛋白5g，CaCO₃ 19g，海盐10g，加水定容至1L，pH 7.0)，28℃培养3d后，抽取各个突变株的基因组DNA，利用检测引物nes27-TestF/R通过PCR检测克隆，如图5(B)所示，阳性结果表示获得nes27敲除双交换突变菌株Δnes27，突变株命名为NES002。When colonies grew on the conjugation transfer plate, use a sterile toothpick to transfer it to ATCC No. 172 medium (20 g starch, 10 g glucose, yeast) containing 35 μg/mL apramycin and 50 μg/mL trimethoprim Extract 5g, Aobox casein 5g, CaCO₃ 19g, sea salt 10g, add water to make up to 1L, pH 7.0), after culturing at 28°C for 3 days, extract the genomic DNA of each mutant strain, use the detection primer nes27-TestF/R to pass PCR The clone was detected, as shown in Figure 5(B), and the positive result indicated that the nes27 knockout double-crossover mutant strain Δnes27 was obtained, and the mutant strain was named NES002.

实施例5：Nenestatin A及其中间体的生物发酵与检测Example 5: Biological fermentation and detection of Nenestatin A and its intermediates

将菌株M.echinospora SCSIO 04089或突变株NES002在ATCC172培养基平板上(淀粉20g，葡萄糖10g，酵母提取物5g，Aobox酪蛋白5g，CaCO₃ 19g，海盐10g，加水定容至1L，pH7.0)活化后，刮取适量孢子接种到含50mL N4培养基(淀粉15g，鱼蛋白胨8g，细菌学蛋白胨5g，甘油6mL，CaCO₃ 2g，KBr 0.2g，海盐10g，加水定容至1L，pH 7.0)的250mL三角烧瓶中，于28℃，200rpm，培养5d后，加入等体积丁酮，超声破碎细胞10min，静置分层，吸取上层丁酮萃取液，旋转蒸发仪蒸干。粗提物使用二甲基亚砜溶解，进行HPLC检测，分析条件为：菲罗门Luna C18(5μm,150×4.6mm)反相柱，流动相A相为10％乙腈水溶液(含0.1％甲酸)，流动相B相为90％乙腈水溶液；流速为1mL/min。检测波长为265nm和480nm。HPLC程序：0-18min，5％Bto 70％B(线性梯度)，19-25min 100％B，27-30min 100％B to 5％B，32min 5％B。Strain M.echinospora SCSIO 04089 or mutant NES002 on ATCC172 medium plate (starch 20g, glucose 10g, yeast extract 5g,_Aobox casein 5g, CaCO3 19g, sea salt 10g, add water to make up to 1L, pH7.0 ) after activation, scrape an appropriate amount of spores and inoculate it into 50mL N4 medium (starch 15g, fish peptone 8g, bacteriological peptone 5g, glycerol 6mL, CaCO₃ 2g, KBr 0.2g, sea salt 10g, add water to make up to 1L, pH 7.0 ) in a 250 mL Erlenmeyer flask, at 28 ° C, 200 rpm, after culturing for 5 d, add an equal volume of butanone, ultrasonically disrupt the cells for 10 min, let stand for stratification, absorb the upper butanone extract, and evaporate to dryness on a rotary evaporator. The crude extract was dissolved in dimethyl sulfoxide and detected by HPLC. The analysis conditions were: Philomena Luna C18 (5μm, 150×4.6mm) reversed-phase column, and the mobile phase A was 10% acetonitrile aqueous solution (containing 0.1% formic acid). ), the mobile phase B was 90% acetonitrile aqueous solution; the flow rate was 1 mL/min. Detection wavelengths are 265 nm and 480 nm. HPLC program: 0-18 min, 5% B to 70% B (linear gradient), 19-25min 100% B, 27-30min 100% B to 5% B, 32min 5% B.

从图5可以看出，Δnes27突变株NES002丧失了产生nenestatin A的能力(图5(C)，(i)和(ii))，而累积一个新的代谢中间产物，经发酵分离纯化，质谱、NMR解析确定了单体化合物的结构式(图1)，该化合物命名为ethyl-dehydrorabelomycin。It can be seen from Figure 5 that the Δnes27 mutant strain NES002 lost the ability to produce nenestatin A (Figure 5(C), (i) and (ii)), but accumulated a new metabolic intermediate, which was isolated and purified by fermentation, mass spectrometry, NMR analysis confirmed the structural formula of the monomeric compound (Figure 1), which was named ethyl-dehydrorabelomycin.

实施例6：Nenestatin A及其中间体的分离鉴定Example 6: Isolation and identification of Nenestatin A and its intermediates

从ATCC172号培养基(淀粉20g，葡萄糖10g，酵母提取物5g，Aobox酪蛋白5g，CaCO₃19g，海盐10g，加水定容至1L，pH 7.0)平板上刮取适量M.echinospora SCSIO 04089的孢子于A1液体培养基中(淀粉10g，酵母提取物4g，细菌学蛋白胨2g，海盐10g，加水定容至1L，pH7.0)，于28℃，200rpm摇床中培养3-4天，待其菌体生长良好作为种子；以10％(v/v)的接种量将种子接入N4培养基中(淀粉15g，鱼蛋白胨8g，细菌学蛋白胨5g，甘油6mL，CaCO₃ 2g，KBr0.2g，海盐10g，加水定容至1L，pH 7.0)，添加5％(m/v)的大孔树脂于28℃，200rpm摇床中培养4天左右，取样检测。Scrape an appropriate amount of spores of M.echinospora SCSIO 04089 from ATCC No. 172 medium (starch 20g, glucose 10g, yeast extract 5g, Aobox casein 5g, CaCO₃ 19g, sea salt 10g, make up to 1L with water, pH 7.0) In A1 liquid medium (starch 10g, yeast extract 4g, bacteriological peptone 2g, sea salt 10g, add water to make up to 1L, pH 7.0), cultivate at 28°C, 200rpm shaker for 3-4 days, wait for The bacteria grow well as seeds; the seeds are inserted into N4 medium at an inoculum of 10% (v/v) (starch 15g, fish peptone 8g, bacteriological peptone 5g, glycerol 6mL, CaCO₃ 2g, KBr 0.2g, 10 g of sea salt, add water to volume to 1 L, pH 7.0), add 5% (m/v) macroporous resin, incubate at 28° C., 200 rpm shaker for about 4 days, and sample for detection.

将发酵液用筛子过滤分离大孔树脂和菌液，树脂用丙酮洗脱，并用旋转蒸发仪浓缩，获得含水浓缩物，再将菌液以4000rpm离心10min，分离发酵液上清与菌体沉淀，菌液上清取样HPLC检测后没有目标产物，舍弃上清。菌体用丙酮浸泡，超声处理后离心，取液体用旋转蒸发仪浓缩，处理三次后将所得水相并入含水浓缩物。将含水浓缩物用丁酮萃取三次，旋转蒸发丁酮萃取液得到粗浸膏(6g)。粗浸膏首先使用正向硅胶(100-200目，100g)梯度洗脱(CHCl₃/MeOH/H₂O，95/5/0.5，90/10/1，V/V，每次600mL)得到2个组分(Fr.1-Fr.2)，其中组分Fr.2经过ODS column(50μm,YMC*GEL ODS-A-HG,30g)在Agela Cheetahtm MP200MPLC工作站上进行洗脱(CH₃OH/H₂O，体积分数为10％-100％)进行1.5小时的梯度洗脱，获得5个亚馏分(Fr.2.1to Fr.2.5)，其中组分Fr2.2经过反向高压制备得到nenestatin A[制备条件：Phenomenex ODS column(250mm×10.0mm i.d.,5μm；Phenomenex Phenomenex ODScolumn(250mm×10.0mm i.d.,5μm；Phenomenex，流动相A,0.1％formic acid in water；流动相B,90％CH₃CN in water；在流动相B为55％的条件下等度洗脱；流速为2.5mL/min，保留时间为13min时得到nenestatin A)，质谱(图7)及NMR(图8和图9)确定其结构(图1)。The fermentation broth was filtered with a sieve to separate the macroporous resin and the bacterial liquid, the resin was eluted with acetone, and concentrated with a rotary evaporator to obtain an aqueous concentrate, and then the bacterial liquid was centrifuged at 4000 rpm for 10 min to separate the fermentation broth supernatant and bacterial cell precipitation, The bacterial liquid supernatant was sampled and detected by HPLC, and there was no target product, and the supernatant was discarded. The bacterial cells were soaked in acetone, centrifuged after ultrasonic treatment, and the liquid was concentrated with a rotary evaporator. After three treatments, the obtained aqueous phase was incorporated into an aqueous concentrate. The aqueous concentrate was extracted three times with butanone, and the butanone extract was rotary evaporated to obtain a crude extract (6 g). The crude extract was first obtained by gradient elution (CHCl₃ /MeOH/H₂ O, 95/5/0.5, 90/10/1, V/V, 600 mL each time) using normal silica gel (100-200 mesh, 100 g) 2 components (Fr.1-Fr.2), wherein the component Fr.2 was eluted (CH₃ OH) on an Agela Cheetahtm MP200MPLC workstation through an ODS column (50 μm, YMC*GEL ODS-A-HG, 30 g). /H₂ O, the volume fraction is 10%-100%) for 1.5 hours gradient elution to obtain 5 sub-fractions (Fr.2.1 to Fr.2.5), wherein the fraction Fr2.2 is prepared by reverse high pressure to obtain nenestatin A[Preparation conditions: Phenomenex ODS column (250mm×10.0mm id, 5μm; Phenomenex Phenomenex ODScolumn (250mm×10.0mm id, 5μm; Phenomenex, mobile phase A, 0.1% formic acid in water; mobile phase B, 90% CH₃ CN in water; isocratic elution at 55% mobile phase B; nenestatin A was obtained at a flow rate of 2.5 mL/min and a retention time of 13 min), mass spectrometry (Fig. 7) and NMR (Fig. 8 and Fig. 9) Determine its structure (Figure 1).

将突变株NES002接种至ATCC172固体培养基(淀粉20g，葡萄糖10g，酵母提取物5g，Aobox酪蛋白5g，CaCO₃ 19g，海盐10g，加水定容至1L，pH 7.0)上，28℃培养7d，接种于A1种子培养基(淀粉10g，酵母提取物4g，细菌学蛋白胨2g，海盐10g，加水定容至1L，pH 7.0)中(50mL培养基/250mL三角瓶)，28℃，200r/min振荡培养3d作为发酵培养的种子，将生长良好的种子以10％(v/v)的接种量接入10L添加5％(m/v)大孔树脂的N4培养基(淀粉15g，鱼蛋白胨8g，细菌学蛋白胨5g，甘油6mL，CaCO₃ 2g，KBr0.2g，海盐10g，加水定容至1L，pH 7.0；)中(200mL培养基/1000mL三角瓶)，28℃，200r/min培养7d后。离心收集大孔树脂后，用丙酮洗脱，旋转蒸发回收丙酮，水相用丁酮萃取，旋转蒸发丁酮萃取液得到6g粗浸膏。粗浸膏用氯仿溶解拌样上硅胶柱(100-200目)，石油醚︰氯仿梯度洗脱(95/5,90/10,v/v,每个比例600mL)，每200mL洗脱收集作为一个馏分，顺序收集了六个馏分，顺序命名为Fr.1-Fr.6，结合HPLC分析，Fr.4上Sephadex LH20柱，收集馏分，结合HPLC分析，合并馏分为L1、L2、L3、L4。L3用HPLC制备50％的CH₃CN︰H₂O(CH₃CN︰H₂O为50︰50)作为流动相等度洗脱(柱子：Phenomenex ODS column(250mm×10.0mm i.d.,5μm；Phenomenex)，流速为2.5mL/min)，制备得到化合物ethyl-dehydrorabelomycin(42mg)(保留时间为23.6min)。质谱(图10)及NMR(图11)确定其结构。The mutant strain NES002 was inoculated into ATCC172 solid medium (starch 20 g, glucose 10 g, yeast extract 5 g, Aobox casein 5 g, CaCO₃ 19 g, sea salt 10 g, added water to make up to 1 L, pH 7.0), and cultured at 28°C for 7 d. Inoculated in A1 seed medium (starch 10g, yeast extract 4g, bacteriological peptone 2g, sea salt 10g, add water to volume to 1L, pH 7.0) (50mL medium/250mL conical flask), 28 ℃, 200r/min shaking Cultivate 3d as the seeds of fermentation culture, insert the well-grown seeds into 10L N4 medium (starch 15g,fish peptone 8g, 5% (m/v) macroporous resin added with 10% (v/v) inoculum amount, Bacteriological peptone 5g, glycerol 6mL, CaCO₃ 2g, KBr 0.2g, sea salt 10g, add water to dilute to 1L, pH 7.0;) (200mL medium/1000mL conical flask), 28 ℃, 200r/min after culturing for 7d. After the macroporous resin was collected by centrifugation, eluted with acetone, the acetone was recovered by rotary evaporation, the aqueous phase was extracted with butanone, and the butanone extract was rotary evaporated to obtain 6 g of crude extract. The crude extract was dissolved in chloroform and mixed with the sample and loaded onto a silica gel column (100-200 mesh), with petroleum ether: chloroform gradient elution (95/5, 90/10, v/v, 600 mL for each ratio), and each 200 mL of elution was collected as One fraction, six fractions were collected sequentially, named Fr.1-Fr.6, combined with HPLC analysis, Fr.4 was placed on a Sephadex LH20 column, the fractions were collected, combined with HPLC analysis, the combined fractions were L1, L2, L3, L4 . L3 was prepared by HPLC with 50%_CH3CN:H2O (50:50 for_CH3CN :_H2O ) as mobile equivalence elution (column: Phenomenex ODS column (250mm x 10.0mm id, 5μm; Phenomenex) , the flow rate was 2.5mL/min), and the compound ethyl-dehydrorabelomycin (42mg) was prepared (retention time was 23.6min). Its structure was confirmed by mass spectrometry (FIG. 10) and NMR (FIG. 11).

序列表sequence listing

<110>中国科学院南海海洋研究所<110> South China Sea Institute of Oceanography, Chinese Academy of Sciences

<120>一种非典型角环素类化合物nenestatin A的生物合成基因簇及其应用<120> Biosynthetic gene cluster of an atypical keratinoid compound nenestatin A and its application

<160>1<160>1

<210>1<210>1

<211>700000<211>700000

<212> DNA<212> DNA

<213>小单孢菌（Micromonospora echinospora）SCSIO 04089<213> Micromonospora echinospora SCSIO 04089

<400> 1<400> 1

cggcaggtag gacaccgact ggacgaaccg cttgagggcg cggttgcgca cttcgttgag 60cggcaggtag gacaccgact ggacgaaccg cttgagggcg cggttgcgca cttcgttgag 60

cagcagcgcg aggacgatcg gcagcgggaa gcagaacagc agggtcagcg cgcccagcac 120cagcagcgcg aggacgatcg gcagcgggaa gcagaacagc agggtcagcg cgcccagcac 120

gagcgtgttg gtgaacacgc tccagaaggt ggggtcggtg aagaacagcc ggaagtaccg 180gagcgtgttg gtgaacacgc tccagaaggt ggggtcggtg aagaacagcc ggaagtaccg 180

cagcccggtc cagtactcgc cgaagatgct gccgcccggc tggaagcgcc ggaacgcgat 240cagcccggtc cagtactcgc cgaagatgct gccgcccggc tggaagcgcc ggaacgcgat 240

cacgttgccg agcatcggca ggtagcggaa cgtcacgaag aacagcagcg gcaggacggc 300cacgttgccg agcatcggca ggtagcggaa cgtcacgaag aacagcagcg gcaggacggc 300

gagcgagtag agctgccagt cccggcgcag ggcccgccgc cagggcgtac gacgactgcg 360gagcgagtag agctgccagt cccggcgcag ggcccgccgc cagggcgtac gacgactgcg 360

cgcggtcggg gctgccaagc tcgtcctccc gcctgaccgt cagatggaac tttccggtaa 420cgcggtcggg gctgccaagc tcgtcctccc gcctgaccgt cagatggaac tttccggtaa 420

tacgtcgtaa gcctttcgga aatttatggg tatcgttatc ggcgtgtcaa gagcgtcgat 480tacgtcgtaa gcctttcgga aatttatggg tatcgttatc ggcgtgtcaa gagcgtcgat 480

tgataactcc ctcgaccagc tccgtcgcta cacgccggcc gtcgtcgagc ccgccgactt 540tgataactcc ctcgaccagc tccgtcgcta cacgccggcc gtcgtcgagc ccgccgactt 540

cgaccggttc tggcgtgcca ccctcaccgc cgccgcagcc accccggtgc tcgtcgacgt 600cgaccggttc tggcgtgcca ccctcaccgc cgccgcagcc accccggtgc tcgtcgacgt 600

ccggcccgaa cccaccgacc tgcggctggt cgacgtgtgg gacgtgacgt tcgccgggtt 660ccggcccgaa cccaccgacc tgcggctggt cgacgtgtgg gacgtgacgt tcgccgggtt 660

cgacggcgaa cctgtccggg cctggtacac ccgtcccgcg ggcgtaccgg cgccgctgcc 720cgacggcgaa cctgtccggg cctggtacac ccgtcccgcg ggcgtaccgg cgccgctgcc 720

ggccgtagtg gagtaccccg gctacgggcg cggccggggc ctgccgggcg agcggctcac 780ggccgtagtg gagtaccccg gctacgggcg cggccggggc ctgccgggcg agcggctcac 780

ctggccggtg gccggttacg cccacctcct ggtggacaac cggggccagg ccgggctgta 840ctggccggtg gccggttacg cccacctcct ggtggacaac cggggccagg ccgggctgta 840

cagccgcggc gacaccccgg acccgcacga cgcgcccggc gggcccagcc ccgccacgcg 900cagccgcggc gacaccccgg acccgcacga cgcgcccggc gggcccagcc ccgccacgcg 900

ggggatcctg tccccggacg actaccacta ccgccgcctg atcaccgacg cggtccgcgc 960ggggatcctg tccccggacg actaccacta ccgccgcctg atcaccgacg cggtccgcgc 960

gatcgacgcc gtccgcgtcc tgcccggcgt ggacccggcc cgcgtcgccg cggtgggcaa 1020gatcgacgcc gtccgcgtcc tgcccggcgt ggacccggcc cgcgtcgccg cggtgggcaa 1020

cagccagggc ggtgggctcg cgctcgcggc ggcgggcctc gtcgacggcc tcgccgccct 1080cagccagggc ggtgggctcg cgctcgcggc ggcgggcctc gtcgacggcc tcgccgccct 1080

cctggtcacc gccccgttcc tgtgcgacat ccagcgcgcc gtcgaactga ccgaggcatc 1140cctggtcacc gccccgttcc tgtgcgacat ccagcgcgcc gtcgaactga ccgaggcatc 1140

cccctacggc gagatcgccc gctacctggc ggtgcaccgc gaggccgagg aggcggtccg 1200cccctacggc gagatcgccc gctacctggc ggtgcaccgc gaggccgagg aggcggtccg 1200

gcgcaccctg tcgtacgtgg acggcgtcac cttcgcccgg cgggccaccg ccccggcgca 1260gcgcaccctg tcgtacgtgg acggcgtcac cttcgcccgg cgggccaccg ccccggcgca 1260

cttcgggatc ggtctgcgcg acgaggtctg cccgccgagc accggcttcg ccgcctacaa 1320cttcgggatc ggtctgcgcg acgaggtctg cccgccgagc accggcttcg ccgcctacaa 1320

ccagtacgcc gccgccggat cgacggtgcc gttacgggag atgcacacgt acccgttcaa 1380ccagtacgcc gccgccggat cgacggtgcc gttacgggag atgcacacgt acccgttcaa 1380

cggccacgag ggcggcgagg ccgtccacgt ccggcggcag ctgcgctggc tcgacgcggt 1440cggccacgag ggcggcgagg ccgtccacgt ccggcggcag ctgcgctggc tcgacgcggt 1440

gatccgtgcc gccgggacgg caggcggctg accggctcac cggatgcccg cacggagcgc 1500gatccgtgcc gccgggacgg caggcggctg accggctcac cggatgcccg cacggagcgc 1500

ccgcacctgg gccggcgcgt ggtggtccgg ctcggggcac cggtgtggcg ggggaccgct 1560ccgcacctgg gccggcgcgt ggtggtccgg ctcggggcac cggtgtggcg ggggaccgct 1560

cacgccgggc ccaggtgcaa ccgtgccagc ttggccgggt cgatcacggc cgtgacggcg 1620cacgccgggc ccaggtgcaa ccgtgccagc ttggccgggt cgatcacggc cgtgacggcg 1620

gtgatccggc cgccggtgac ggcgaacgcg aggatcgaga gcggcgtgcc gtccgcgcgc 1680gtgatccggc cgccggtgac ggcgaacgcg aggatcgaga gcggcgtgcc gtccgcgcgc 1680

caggagacaa ctccgggcag gccgtccacc agcgcggtcc gggtcgacgc ggcctgggcg 1740caggagacaa ctccgggcag gccgtccacc agcgcggtcc gggtcgacgc ggcctgggcg 1740

gcggcggccc gcgcgccggc ggcgaccctg gtggcaccga tggtgaccac cgcgtcgccg 1800gcggcggccc gcgcgccggc ggcgaccctg gtggcaccga tggtgaccac cgcgtcgccg 1800

ggaccgtcga cggtgagctt cacgtcggga tcgagcacgc gaagcagacc ttcgaagtct 1860ggaccgtcga cggtgagctt cacgtcggga tcgagcacgc gaagcagacc ttcgaagtct 1860

ccaccgcgag ccgccgccag gaaggcggag accacctcgc gttgttcccg gcccgggccg 1920ccaccgcgag ccgccgccag gaaggcggag accacctcgc gttgttcccg gcccgggccg 1920

gtcgggcgtt cggtcgcctg gaccttcctg cgggcacggc tggcgagcat cttggtggcg 1980gtcgggcgtt cggtcgcctg gaccttcctg cgggcacggc tggcgagcat cttggtggcg 1980

tcggtggact tgccgaggat gcggccgatc tcgtcgaacg gcaccgcgaa caggtcgtgg 2040tcggtggact tgccgaggat gcggccgatc tcgtcgaacg gcaccgcgaa caggtcgtgg 2040

agcacgaatg ccagccgctc gctcgggcgc agtgaatcga gaacgacgag gagcgcgagc 2100agcacgaatg ccagccgctc gctcgggcgc agtgaatcga gaacgacgag gagcgcgagc 2100

ccgaccgagt cggcgagcac cgcgtcgtcg tccggggcgt gtgcgtcgtc gaacgtcacc 2160ccgaccgagt cggcgagcac cgcgtcgtcg tccggggcgt gtgcgtcgtc gaacgtcacc 2160

acgaggtcgg gaaggtggat gtcgaaagag gcctcggggc gggcctggcg cgaccgcagg 2220acgaggtcgg gaaggtggat gtcgaaagag gcctcggggc gggcctggcg cgaccgcagg 2220

acgtcgaggc tgagccggcc gaccactgtg gtgagccatc cggcgaggtt gtcgatggtg 2280acgtcgaggc tgagccggcc gaccactgtg gtgagccatc cggcgaggtt gtcgatggtg 2280

tccgcgtcct ggcgggcgag tctcagccag gcttcctgga ccacgtcctc ggcgtcggcg 2340tccgcgtcct ggcgggcgag tctcagccag gcttcctgga ccacgtcctc ggcgtcggcg 2340

tgcgacccga gtacgcggta ggcgaccgcg cgcagccggt cgcggtggga ctcgaatgcc 2400tgcgacccga gtacgcggta ggcgaccgcg cgcagccggt cgcggtggga ctcgaatgcc 2400

gttgccgccg agtccgtcga gccgctgtcg accatgttgt taccttccac gattccgctt 2460gttgccgccg agtccgtcga gccgctgtcg accatgttgt taccttccac gattccgctt 2460

cgtcagaggt gatgacgggt ccgggcgggc ccgggtaacc gctgaaggag cagggactga 2520cgtcagaggt gatgacgggt ccgggcgggc ccgggtaacc gctgaaggag cagggactga 2520

tggaaccacg cctcaagagt gcgatgaacc cggatctgat gactgcggtc cagcacctcc 2580tggaaccacg cctcaagagt gcgatgaacc cggatctgat gactgcggtc cagcacctcc 2580

acaaggcgat agccgccggg ggtgtcgagc cgcgcctgct gtcgctggtc cacctccgcg 2640acaaggcgat agccgccggg ggtgtcgagc cgcgcctgct gtcgctggtc cacctccgcg 2640

ccagtcagat caacggctgt gcgccgtgcg tcttcgcgtc cgtctcgggg gcgaagaagg 2700ccagtcagat caacggctgt gcgccgtgcg tcttcgcgtc cgtctcgggg gcgaagaagg 2700

ccggcgagac ggacgagcgg ctgcaccacg tggtcgcgtg gcgggagacg ccgttcttca 2760ccggcgagac ggacgagcgg ctgcaccacg tggtcgcgtg gcgggagacg ccgttcttca 2760

gcgaacagga gcgggcggcg ctcgcgctga ccgaggccgc cacccgcatt caggacggtg 2820gcgaacagga gcgggcggcg ctcgcgctga ccgaggccgc cacccgcatt caggacggtg 2820

cccccggcgt gaccgatcag gtctgggacg ccgccgccga acacttcagc gaggaggagt 2880cccccggcgt gaccgatcag gtctgggacg ccgccgccga acacttcagc gaggaggagt 2880

tgaacgcgat cgtcctggag atcgccatga ccaacttctt caaccggatc aaccactcca 2940tgaacgcgat cgtcctggag atcgccatga ccaacttctt caaccggatc aaccactcca 2940

tccgggaaca ggccggcaag acctggtgag tcgccccgca ccatcacctc accgccgccg 3000tccgggaaca ggccggcaag acctggtgag tcgccccgca ccatcacctc accgccgccg 3000

atgaaggcga agacattcct gattcgacct cggtttctgg caatacggat cgtgggtacc 3060atgaaggcga agacattcct gattcgacct cggtttctgg caatacggat cgtgggtacc 3060

gaaaagaaag tgatcaggaa tagagaaggc gtggcacacc gggtgcgact agcctgagga 3120gaaaagaaag tgatcaggaa tagagaaggc gtggcacacc gggtgcgact agcctgagga 3120

gaacggtgac gagagagagg agcggtacct aagtgagcac cacgtcgaaa gccagtaagg 3180gaacggtgac gagagagagg agcggtacct aagtgagcac cacgtcgaaa gccagtaagg 3180

tcggcaagca gagcaagtct tccgatgagg gattctccga ggtcgagcga gccgccatca 3240tcggcaagca gagcaagtct tccgatgagg gattctccga ggtcgagcga gccgccatca 3240

aggaccgcgc ggccgagttg aagtcggagg ctcgccgcag caagagcgcg aacaaggcgg 3300aggaccgcgc ggccgagttg aagtcggagg ctcgccgcag caagagcgcg aacaaggcgg 3300

cggccgacga gaaggacgtg ctcgcgaaga tcgccgagat ggggcagcct gaccggaagg 3360cggccgacga gaaggacgtg ctcgcgaaga tcgccgagat ggggcagcct gaccggaagg 3360

tggccgagcg cctgcacgcc atcatcacgg aaaccgctcc cgaactggcc ccgaagctgt 3420tggccgagcg cctgcacgcc atcatcacgg aaaccgctcc cgaactggcc ccgaagctgt 3420

ggtacggcca gcccgcctac gcccgcggcg ggaaggtggt gtgcttcttc cgcagcggca 3480ggtacggcca gcccgcctac gcccgcggcg ggaaggtggt gtgcttcttc cgcagcggca 3480

aggtggacaa ggaacgctac tcgtcgttcg ggttcacgac ggaagcgaag ctcgacgagg 3540aggtggacaa ggaacgctac tcgtcgttcg ggttcacgac ggaagcgaag ctcgacgagg 3540

accatggcct ctggccgacg tccttcgctc tcaccgagct cagcgacaag ggtgaggcga 3600accatggcct ctggccgacg tccttcgctc tcaccgagct cagcgacaag ggtgaggcga 3600

cgatcaagaa gcttctgaag aaggctatgg gttgatttcc gcggtcgaaa acagcgcggg 3660cgatcaagaa gcttctgaag aaggctatgg gttgatttcc gcggtcgaaa acagcgcggg 3660

cgacccagcg tcatagtccg gccgagcagc gacagctgct cggccggccc gcgcttccgg 3720cgacccagcg tcatagtccg gccgagcagc gacagctgct cggccggccc gcgcttccgg 3720

cccgtctctg cagcccgtgt gcccacggcg tcgcgcgacg ccgtgggcac acgggtttcg 3780cccgtctctg cagcccgtgt gcccacggcg tcgcgcgacg ccgtgggcac acgggtttcg 3780

tcgtcggtca ggcgtctcgg cgcttgagca ggacagctgc gagcgcgagc agcgcggcgg 3840tcgtcggtca ggcgtctcgg cgcttgagca ggacagctgc gagcgcgagc agcgcggcgg 3840

tccacaggca gaacactccg aagccctgcc acggcgtgag cagatcgccg gggagcttcg 3900tccacaggca gaacactccg aagccctgcc acggcgtgag cagatcgccg gggagcttcg 3900

tggcctgccc gatgaggatt ccggcctcgg tgggcaggta cgcgtgggcg tactcaccga 3960tggcctgccc gatgaggatt ccggcctcgg tgggcaggta cgcgtgggcg tactcaccga 3960

ccttgccggg cagcagctgg accagcggag ccagcacgag cacgaacatg atcacgctgg 4020ccttgccggg cagcagctgg accagcggag ccagcacgag cacgaacatg atcacgctgg 4020

tgatgccggc cgcggtgcgc cggacgatgg cgccgatggc gaacgcgaac aggctcagca 4080tgatgccggc cgcggtgcgc cggacgatgg cgccgatggc gaacgcgaac aggctcagca 4080

cggccaggta cagcgccgtg cccaccacgg ccctcgccac accggggtcg cccagcgaga 4140cggccaggta cagcgccgtg cccaccacgg ccctcgccac accggggtcg cccagcgaga 4140

cgtcgacctt gctggccagc atcgaggccc cgacgaagaa ggaggcgaac gccgccacga 4200cgtcgacctt gctggccagc atcgaggccc cgacgaagaa ggaggcgaac gccgccacga 4200

gactcacggc caggaccagc aggcacaaca ccacggcctt ggcggcgaga agcggcatcc 4260gactcacggc caggaccagc aggcacaaca ccacggcctt ggcggcgaga agcggcatcc 4260

gccgcggtac ggcgaggagg ctggcacgga tcatgccggt cgagtactcc gacgcgatcg 4320gccgcggtac ggcgaggagg ctggcacgga tcatgccggt cgagtactcc gacgcgatcg 4320

ccatgacacc caggacgcag atcgccggct ggctgatcag gaagccgctg ccgaggatga 4380ccatgacacc caggacgcag atcgccggct ggctgatcag gaagccgctg ccgaggatga 4380

cggcggcggg atccgaggcg gccagggccc ggtcgctctc ggtcatctga tcccacgagg 4440cggcggcggg atccgaggcg gccagggccc ggtcgctctc ggtcatctga tcccacgagg 4440

cgacggtcag gccgacgaac agcgccgtga aggccggata cagcaccgcg agcagggcga 4500cgacggtcag gccgacgaac agcgccgtga aggccggata cagcaccgcg agcagggcga 4500

gcgaccatcc ggtcgaccgc accgacttca gcttcgtcca ttcggagagc atcacctggc 4560gcgaccatcc ggtcgaccgc accgacttca gcttcgtcca ttcggagagc atcacctggc 4560

cgaacccgga gcgactggcc accggtgtcg tggtgacgtc agcagtggcc gtggtcatcg 4620cgaacccgga gcgactggcc accggtgtcg tggtgacgtc agcagtggcc gtggtcatcg 4620

ggcagctcct cgggagttgg gggcgacggt ttcgcgggtg gatccggtgc caccggtacc 4680ggcagctcct cgggagttgg gggcgacggt ttcgcgggtg gatccggtgc caccggtacc 4680

ggtgtactcg acgctctcgc gcgtcagttc catgaacgcc tcttccaatg tgcttcgctg 4740ggtgtactcg acgctctcgc gcgtcagttc catgaacgcc tcttccaatg tgcttcgctg 4740

cggcacgacc tcgtgcagga cgtgcccccc ggcggaggcg atctcgccga tccgggccgc 4800cggcacgacc tcgtgcagga cgtgcccccc ggcggaggcg atctcgccga tccgggccgc 4800

cgtcatgtcc tggacactca gttcgccgtc gccgttgcgg gtcacctcgc cgccctcacg 4860cgtcatgtcc tggacactca gttcgccgtc gccgttgcgg gtcacctcgc cgccctcacg 4860

cgtgatcgcc tcggccagca ggtcggcgtg cgggctgcgg accagcaccg tctgcttcct 4920cgtgatcgcc tcggccagca ggtcggcgtg cgggctgcgg accagcaccg tctgcttcct 4920

gccgcgttcg atgaactcct gggtggggca gtcggcgacc aaccggccgc gtccgatgat 4980gccgcgttcg atgaactcct gggtggggca gtcggcgacc aaccggccgc gtccgatgat 4980

gatgagatgc tcggcggtca ccgccatctc gttcatcagg tggctggaga cgaacacggt 5040gatgagatgc tcggcggtca ccgccatctc gttcatcagg tggctggaga cgaacacggt 5040

tcgcccctcg gccgccagtt tcctcatcag ggtgcgaatc cacaggatgc cctcagggtc 5100tcgcccctcg gccgccagtt tcctcatcag ggtgcgaatc cacaggatgc cctcagggtc 5100

gagcccgttg accggctcgt ccaggatcag cacggacggg tcgccgagca gggcggcggc 5160gagcccgttg accggctcgt ccaggatcag cacggacggg tcgccgagca gggcggcggc 5160

cagcccgaga cgctgcccca taccgaggga gaagccgccg gcccgcttgc gtgctacctc 5220cagcccgaga cgctgcccca taccgaggga gaagccgccg gcccgcttgc gtgctacctc 5220

cgtcagcccg acgaggtcga ggacctcgtt cacccgcttc ttgccgatgc cctgcgactg 5280cgtcagcccg acgaggtcga ggacctcgtt cacccgcttc ttgccgatgc cctgcgactg 5280

ggcgaggcag agcaggtggt tgtacgcgct gcggccgggg tgcacggcct tggcctccaa 5340ggcgaggcag agcaggtggt tgtacgcgct gcggccgggg tgcacggcct tggcctccaa 5340

caacgagccg acgacggtca gcggcgtggc gagttcgtgg tactgcttgc cgtcgatggt 5400caacgagccg acgacggtca gcggcgtggc gagttcgtgg tactgcttgc cgtcgatggt 5400

ggcgttgccg ccgtcgggac ggtcgaggcc cagcaacagg cgcatcgtcg tcgattttcc 5460ggcgttgccg ccgtcgggac ggtcgaggcc cagcaacagg cgcatcgtcg tcgattttcc 5460

ggcgccgttg ggcccgagga atccggtcac cttgccgggg ccgacggtga aggacagatc 5520ggcgccgttg ggcccgagga atccggtcac cttgccgggg ccgacggtga aggacagatc 5520

cttcaccgcg gtcttgtcgc cgtagcgctt ggtaaggttc ctggcttcga tcatccgttc 5580cttcaccgcg gtcttgtcgc cgtagcgctt ggtaaggttc ctggcttcga tcatccgttc 5580

cgtcctggat taggcgatga gtgggacaaa gcgagggagt cggaatcagt cgcgttcgca 5640cgtcctggat taggcgatga gtgggacaaa gcgagggagt cggaatcagt cgcgttcgca 5640

ttcgccccgg gaacaacgct cccgagcagt catcgggaac gcggttgggc gaaacgcagc 5700ttcgccccgg gaacaacgct cccgagcagt catcgggaac gcggttgggc gaaacgcagc 5700

gcattgccgg cgggatcctg gacggcgcag tcgcgaacgc cccagggctg atccgtcggt 5760gcattgccgg cgggatcctg gacggcgcag tcgcgaacgc cccagggctg atccgtcggt 5760

tcctgcgtga ccgtcgctcc cgcttcgcgg aggcgctcga acgtggcgtc gcagtcgtcg 5820tcctgcgtga ccgtcgctcc cgcttcgcgg aggcgctcga acgtggcgtc gcagtcgtcg 5820

gtacggaaca cgagaccggg cagcaacccc ttcgccagaa ggcttgcgag ggcctgctgg 5880gtacggaaca cgagaccggg cagcaacccc ttcgccagaa ggcttgcgag ggcctgctgg 5880

tcggcgggcg aggtctccgg atcgacgaca agggactgga ggacgatgtc ggcgtcaggc 5940tcggcgggcg aggtctccgg atcgacgaca agggactgga ggacgatgtc ggcgtcaggc 5940

tgtgcggccg aaccgagagt cacccagcgc agtccctcga acgctatgtc cttgagcacc 6000tgtgcggccg aaccgagagt cacccagcgc agtccctcga acgctatgtc cttgagcacc 6000

tcgaaaccga ggacatcccg gtagaaggca attgccttct cgtggtcgtc cacggcgacg 6060tcgaaaccga ggacatcccg gtagaaggca attgccttct cgtggtcgtc cacggcgacg 6060

aagcactgtg aaaccttgac tcccatgcgt gtcacgctac tgacagacgg tgcccggttg 6120aagcactgtg aaaccttgac tcccatgcgt gtcacgctac tgacagacgg tgcccggttg 6120

cttctcgatt cctgacgggt gtcgggccga tccggacggg aaggcccgtg tgccgacggt 6180cttctcgatt cctgacgggt gtcgggccga tccggacggg aaggcccgtg tgccgacggt 6180

tccataagag cgtccttccg tgcgccggtg agcctcaggc cttgacgaat tcgaccagga 6240tccataagag cgtccttccg tgcgccggtg agcctcaggc cttgacgaat tcgaccagga 6240

cccgggcgag gatttcggtc tccacccggt gccaggatcc gggcagttcg cgtccggagc 6300cccgggcgag gatttcggtc tccacccggt gccaggatcc gggcagttcg cgtccggagc 6300

cgttgggcag ggccttcgcc gtcgcctgcg ccgccgcccg cagccagtca ctggtgccgt 6360cgttgggcag ggccttcgcc gtcgcctgcg ccgccgcccg cagccagtca ctggtgccgt 6360

cgctgttgac gaccaacgtc tccgtccgga gggccgcgag ccgctcgacc gggacgtcga 6420cgctgttgac gaccaacgtc tccgtccgga gggccgcgag ccgctcgacc gggacgtcga 6420

aatcaccgca gattgccgtg tcgtacgcga gcgtgtgggc gttcgcctcg ttcgtcgccc 6480aatcaccgca gattgccgtg tcgtacgcga gcgtgtgggc gttcgcctcg ttcgtcgccc 6480

acagcgggcc ctgccgccac tggctgatcg cctcgggcgg cagtcccatc agcaccgtga 6540acagcgggcc ctgccgccac tggctgatcg cctcgggcgg cagtcccatc agcaccgtga 6540

ggaagtactc cgccgcttcg ccgcgcctgc cgtcggcaac aagcgcccgg agcgcgtcgg 6600ggaagtactc cgccgcttcg ccgcgcctgc cgtcggcaac aagcgcccgg agcgcgtcgg 6600

cggcgtcgag gggcggcttc gggtgaccgt cgacccggaa gtacggctcg tgcagtgcga 6660cggcgtcgag gggcggcttc gggtgaccgt cgacccggaa gtacggctcg tgcagtgcga 6660

gcttcgtgat cgccagaccc gccatcgccg cctccagcgc gaggttcgcg ccgcacgaac 6720gcttcgtgat cgccagaccc gccatcgccg cctccagcgc gaggttcgcg ccgcacgaac 6720

cggcgaacac cacagcgctg ccgccggcct cctcgatcac cgcggcgaga tcctcgatct 6780cggcgaacac cacagcgctg ccgccggcct cctcgatcac cgcggcgaga tcctcgatct 6780

cgcgctcgac cgcgtagacg ggcgagtcac cgctctgtcc ccgtcctcgg cggtcgtaga 6840cgcgctcgac cgcgtagacg ggcgagtcac cgctctgtcc ccgtcctcgg cggtcgtaga 6840

cgaacgtggt gcagtacggc gccagctcgg gcacgagcgc gtcgaagatc gtgtgatcgc 6900cgaacgtggt gcagtacggc gccagctcgg gcacgagcgc gtcgaagatc gtgtgatcgc 6900

gaaacgcgcc gttcagcaag acgatcggtg gcccttcgcc ggctcgttca taggcgatcg 6960gaaacgcgcc gttcagcaag acgatcggtg gcccttcgcc ggctcgttca taggcgatcg 6960

tcgtaccgtc ccgcgaaacg accttgcgca tggcgctctc ctcagtgggc ccgcacggat 7020tcgtaccgtc ccgcgaaacg accttgcgca tggcgctctc ctcagtgggc ccgcacggat 7020

gaatccgcag ccaacgtgcg gctggctggt tgacgtccgg tcgaggagcg gtcgtgcctg 7080gaatccgcag ccaacgtgcg gctggctggt tgacgtccgg tcgaggagcg gtcgtgcctg 7080

acgcggtccg aatcgccgcg ccggcggcga ggtctacgag gcgatggcgg tgacgccgcc 7140acgcggtccg aatcgccgcg ccggcggcga ggtctacgag gcgatggcgg tgacgccgcc 7140

ctccaacccc ggatcgccgg cgaggatcgc ggccaggagt cgttgcaccc gcgggcctgg 7200ctccaacccc ggatcgccgg cgaggatcgc ggccaggagt cgttgcaccc gcgggcctgg 7200

ttcgacgccg agctcgtcgt tcagcacggc acgaaggcgg tgatagacct gcagcgatcg 7260ttcgacgccg agctcgtcgt tcagcacggc acgaaggcgg tgatagacct gcagcgatcg 7260

accgacgtgg ccggcgcggt acatggctgt catcagcagg gcacagaggt tctcgttcat 7320accgacgtgg ccggcgcggt acatggctgt catcagcagg gcacagaggt tctcgttcat 7320

cggattccgg gccgtcagga gcgtcagctc ggccagcagg tcggagtgcc gcccgagagc 7380cggattccgg gccgtcagga gcgtcagctc ggccagcagg tcggagtgcc gcccgagagc 7380

gaggtcggcc tcgatccggc gttccagcgc gccgagccgg ctctcctcca gcgaagccgc 7440gaggtcggcc tcgatccggc gttccagcgc gccgagccgg ctctcctcca gcgaagccgc 7440

ctcgatctcg agtatcggcc ccattcggac gtcgaccagt gccgagccgc gccatacctg 7500ctcgatctcg agtatcggcc ccattcggac gtcgaccagt gccgagccgc gccatacctg 7500

cagcgcgcga ctgaattcat gggaagcccg gcggtgatcg tctgattcgg ccgcttcgcg 7560cagcgcgcga ctgaattcat gggaagcccg gcggtgatcg tctgattcgg ccgcttcgcg 7560

accggcgacg agatggcggt tgtagacctc gacgtccgtt tcgcaggcgt ggtcctcgag 7620accggcgacg agatggcggt tgtagacctc gacgtccgtt tcgcaggcgt ggtcctcgag 7620

caggtaaccg ttgtggcggg tgctgatgat ctgccgtgcc cgcagattcc cgcagccggc 7680caggtaaccg ttgtggcggg tgctgatgat ctgccgtgcc cgcagattcc cgcagccggc 7680

ggcggcaaga gcgttgcgca actgcagaat gtacgtctgc aacgtggtgg cgtggcttcg 7740ggcggcaaga gcgttgcgca actgcagaat gtacgtctgc aacgtggtgg cgtggcttcg 7740

tggtgggcgg tccccccaca gttcttcgat aagtgtcggc acggtgacga cgcggcccgc 7800tggtgggcgg tccccccaca gttcttcgat aagtgtcggc acggtgacga cgcggcccgc 7800

attgagcgcg agaagagcga gaacctgtcg ttgtttcggt gcgcttgccg tcaccgccgt 7860attgagcgcg agaagagcga gaacctgtcg ttgtttcggt gcgcttgccg tcaccgccgt 7860

ccggcagaaa cgtagggaca gtgggccgag cagcccgata ctgagtggct tctgattctt 7920ccggcagaaa cgtagggaca gtgggccgag cagcccgata ctgagtggct tctgattctt 7920

ctccatataa tctcctgccc cgtttcggat tgctgtggcc gagctctccg tcgtgacata 7980ctccatataa tctcctgccc cgtttcggat tgctgtggcc gagctctccg tcgtgacata 7980

acaaatgctt gaccgatgcg tcttcggtac acaacgattt tattcacgcg aatgtgaaaa 8040acaaatgctt gaccgatgcg tcttcggtac acaacgattt tattcacgcg aatgtgaaaa 8040

taggcgcgat tggggcacaa ggcccggctc ggcgccgatg gtaacgggtg caagtttttt 8100taggcgcgat tggggcacaa ggcccggctc ggcgccgatg gtaacgggtg caagtttttt 8100

actcgatgag tctcccccta caagatcgaa taactggatt actgcacgcg attacgcatt 8160actcgatgag tctcccccta caagatcgaa taactggatt actgcacgcg attacgcatt 8160

gtggtgtgat ccggtggtcc cggggaactg cgggtttatc gtacccgacc gcttacctga 8220gtggtgtgat ccggtggtcc cggggaactg cgggtttatc gtacccgacc gcttacctga 8220

tgcgctgcaa aagttggggt gggatcgcgt atccggaggc tgagctgcga gaagtcgatg 8280tgcgctgcaa aagttggggt gggatcgcgt atccggaggc tgagctgcga gaagtcgatg 8280

tgtcgagatt tgggacgcgt cctctttaga ctgcttcgac ggactcggcg cccgccggtt 8340tgtcgagatt tgggacgcgt cctctttaga ctgcttcgac ggactcggcg cccgccggtt 8340

cccgcgctgc tgccggggtg cggccgatgg cctccggggg ccgcgtcgag gcaacggaaa 8400cccgcgctgc tgccggggtg cggccgatgg cctccggggg ccgcgtcgag gcaacggaaa 8400

gcgtccgctg gggccagctc gcggcgagcc tcatcggtgc ccggtccgcg tcgatgcagg 8460gcgtccgctg gggccagctc gcggcgagcc tcatcggtgc ccggtccgcg tcgatgcagg 8460

ccgtgatcca caccggtggc gttcgtcaca aatgctcgtc aggtgccctg ggcaaccgaa 8520ccgtgatcca caccggtggc gttcgtcaca aatgctcgtc aggtgccctg ggcaaccgaa 8520

ttcgcgccag gcggcgggtc ggccgacgaa cccggcggcg accgcgtcgg agggccggcg 8580ttcgcgccag gcggcgggtc ggccgacgaa cccggcggcg accgcgtcgg agggccggcg 8580

gatcacggtc tgcaacgtcc gccccgcttc cggccgtacc accgagccgc cgcagcgact 8640gatcacggtc tgcaacgtcc gccccgcttc cggccgtacc accgagccgc cgcagcgact 8640

tcgcagcagc gtcgtttccg ggccggttcg acttccggga agaccagcgc cggtttccgg 8700tcgcagcagc gtcgtttccg ggccggttcg acttccggga agaccagcgc cggtttccgg 8700

cgaatagggg gacgcgccgc gccaggaatc gggcggtaac cggtgaccct gttgcgcgaa 8760cgaatagggg gacgcgccgc gccaggaatc gggcggtaac cggtgaccct gttgcgcgaa 8760

ccgggccggt tcgcccctca tctccgcgga gccacgaatt gtcgcgggac gtggtcggtc 8820ccgggccggt tcgcccctca tctccgcgga gccacgaatt gtcgcgggac gtggtcggtc 8820

gatgaggctg ctccggcctg atgcgccact ccggggatcc ggtgaatggg ccggcgaaag 8880gatgaggctg ctccggcctg atgcgccact ccggggatcc ggtgaatggg ccggcgaaag 8880

gcgtgccacc gatcatcggt gccgactgcc ggttcgaggt ccgacagatc cgtgattcgc 8940gcgtgccacc gatcatcggt gccgactgcc ggttcgaggt ccgacagatc cgtgattcgc 8940

cgcctcgcac cgtcgcgtga gggcggcgcg gggtgttgcg aagattcgcg ggctggctgt 9000cgcctcgcac cgtcgcgtga gggcggcgcg gggtgttgcg aagattcgcg ggctggctgt 9000

ccgtaccggc gctgccgagc ggaaatggcg ggcctcctgg ctcgcggccc ggcctcacgt 9060ccgtaccggc gctgccgagc ggaaatggcg ggcctcctgg ctcgcggccc ggcctcacgt 9060

cagcggtacc ggagtgccga attggtttcc ggcgcttcgg cccgggattt gctgctcgga 9120cagcggtacc ggagtgccga attggtttcc ggcgcttcgg cccgggattt gctgctcgga 9120

gccgggcacc agcggtgctc gagctgatcc cgacggcccg aggggatgct gacggctgcc 9180gccgggcacc agcggtgctc gagctgatcc cgacggcccg aggggatgct gacggctgcc 9180

acggatgtag tgaggtgccc atggaaaatg cagtcgaacg tgccgtcgag tatctctgga 9240acggatgtag tgaggtgccc atggaaaatg cagtcgaacg tgccgtcgag tatctctgga 9240

agcactacga ccgaccgctg tcgctgaccg aggtggcgga gagcgtccgc ctgagccgtt 9300agcactacga ccgaccgctg tcgctgaccg aggtggcgga gagcgtccgc ctgagccgtt 9300

tctacttcgc ccgcctgttc cgggacacca ccggaatcac ccccgggcgc tttctggccg 9360tctacttcgc ccgcctgttc cgggacacca ccggaatcac ccccgggcgc tttctggccg 9360

ccattcgcat acaccacgcc aaacggctcc tcattgacac gtcgatgcgg ataaccgatg 9420ccattcgcat acaccacgcc aaacggctcc tcattgacac gtcgatgcgg ataaccgatg 9420

tctccgtgtc ggtgggttac aacagcctgg ggtcgttcag caattgcttc acctccagcg 9480tctccgtgtc ggtgggttac aacagcctgg ggtcgttcag caattgcttc acctccagcg 9480

tcggcctgtc acccggccgt ttccggcgtc tgtcgcagat cgacggcatc gagctgccgg 9540tcggcctgtc acccggccgt ttccggcgtc tgtcgcagat cgacggcatc gagctgccgg 9540

gcgcgttgcc ggccccgcac ggtccctcgg gcgcggtcgc ggggacgatc agcctgccgg 9600gcgcgttgcc ggccccgcac ggtccctcgg gcgcggtcgc ggggacgatc agcctgccgg 9600

aggggcacgg gaacgcgcgc gtgtatctgg gcgccttccg gacgcagacc gtgaagcacc 9660aggggcacgg gaacgcgcgc gtgtatctgg gcgccttccg gacgcagacc gtgaagcacc 9660

cggcggccgc cgcggtgatc gtcgacgttc ccggtggccg gccggcgtgc taccgcctgc 9720cggcggccgc cgcggtgatc gtcgacgttc ccggtggccg gccggcgtgc taccgcctgc 9720

cgcacgttcc cgagggcacc tggatcgtgc acgcggtggc ggtggccgac ggcgccggac 9780cgcacgttcc cgagggcacc tggatcgtgc acgcggtggc ggtggccgac ggcgccggac 9780

tcgacgcgcg gcggccgcac gccccactgg tcggaatgca cccgtcggtg acggtgaccg 9840tcgacgcgcg gcggccgcac gccccactgg tcggaatgca cccgtcggtg acggtgaccg 9840

cgggtggagt gaccagcgcc gcggtccgtc tgcgtcgccg ccggccgatc gacccgccgg 9900cgggtggagt gaccagcgcc gcggtccgtc tgcgtcgccg ccggccgatc gacccgccgg 9900

tgctgctcgc cctgcccgac ctggtgccgc cgcctgcgcc ggcaccgctc accgggtgcc 9960tgctgctcgc cctgcccgac ctggtgccgc cgcctgcgcc ggcaccgctc accgggtgcc 9960

cggccgtcac ggcacgccag gcgccggccg cgtggcagga ggatcgacgt gggctggcgc 10020cggccgtcac ggcacgccag gcgccggccg cgtggcagga ggatcgacgt gggctggcgc 10020

tggccgcgcc ccgtcccggg ttgtcgtagc gggcccctcc ggcacgaaca cggtgtgagg 10080tggccgcgcc ccgtcccggg ttgtcgtagc gggcccctcc ggcacgaaca cggtgtgagg 10080

tgccgctttc aactgcgaat attgatgcag ttgaacgtca ccgagccggg acggcggtgc 10140tgccgctttc aactgcgaat attgatgcag ttgaacgtca ccgagccggg acggcggtgc 10140

tcccggccgg tcgggacacg gccctccctc agcggagagg gctcgcgccg atccggcacc 10200tcccggccgg tcgggacacg gccctccctc agcggagagg gctcgcgccg atccggcacc 10200

tcatcgaacc atgctcggcg cggcccggtc cgtcaagccc gcggcttcct ctgtcggcgg 10260tcatcgaacc atgctcggcg cggcccggtc cgtcaagccc gcggcttcct ctgtcggcgg 10260

tcgagtccgg caacccgggg ggcaacgtgg aggatgccgg ccgatcgcgg gccactgcac 10320tcgagtccgg caacccgggg ggcaacgtgg aggatgccgg ccgatcgcgg gccactgcac 10320

tatggccgag acacagacgg attgccccat cgggactgtc tctccgcggc ccgtcgcaca 10380tatggccgag acacagacgg attgccccat cgggactgtc tctccgcggc ccgtcgcaca 10380

gcggtgggga aagggctggg gccaccactg cccagcggcc ttcgcatcgt cggcaccgcc 10440gcggtgggga aagggctggg gccaccactg cccagcggcc ttcgcatcgt cggcaccgcc 10440

gcaagcggaa cacgcgagcc agaaagtggt gatcgaccgt gacctcagcc gtactgctca 10500gcaagcggaa cacgcgagcc agaaagtggt gatcgaccgt gacctcagcc gtactgctca 10500

gacctcgctc cggcaccgcc ctcacggaac ggctggcgcg ttcggcggcg gcgaccgagg 10560gacctcgctc cggcaccgcc ctcacggaac ggctggcgcg ttcggcggcg gcgaccgagg 10560

gcgccggcat caagacggcc gacgtgccgg gctggctggc cgccgagacg gtggcctgcg 10620gcgccggcat caagacggcc gacgtgccgg gctggctggc cgccgagacg gtggcctgcg 10620

gtaccagggt gcgccggatt cccttcgctg agctggacgg gtggtcgttc agtccgaccg 10680gtaccagggt gcgccggatt cccttcgctg agctggacgg gtggtcgttc agtccgaccg 10680

gcgggaacct gcgacaccgc agcggacgct tcttctccgt cgaaggcctg catgtcgccc 10740gcgggaacct gcgacaccgc agcggacgct tcttctccgt cgaaggcctg catgtcgccc 10740

ggccggacat cggcaccgag tggcagcagc cgatcatcat gcaaccggag gtcggcatcc 10800ggccggacat cggcaccgag tggcagcagc cgatcatcat gcaaccggag gtcggcatcc 10800

tcggcattct ggccaaggag ttcgacggga cgctgcactt cctcatgcag gcgaaggtgg 10860tcggcattct ggccaaggag ttcgacggga cgctgcactt cctcatgcag gcgaaggtgg 10860

agccgggcaa ccccaacctg gtccagctgt cgcccaccgt gcaggcgaca cgtagcaact 10920agccgggcaa ccccaacctg gtccagctgt cgcccaccgt gcaggcgaca cgtagcaact 10920

acacgaaggc ccacggcggg gcggcggtga agtacctgga gtactttctc cgtcccgacc 10980acacgaaggc ccacggcggg gcggcggtga agtacctgga gtactttctc cgtcccgacc 10980

cccggatggt gatcaccgac gtgctgcagt cggagcacgg tgcgtggttc taccgcaagc 11040cccggatggt gatcaccgac gtgctgcagt cggagcacgg tgcgtggttc taccgcaagc 11040

gcaaccgcaa catgatcgtc gaggtcgagg acgacgtgcc ggccgacgag aacttccggt 11100gcaaccgcaa catgatcgtc gaggtcgagg acgacgtgcc ggccgacgag aacttccggt 11100

ggctcaccct gggccagctc ggacggcttc tccagcacga caacgtcgtg aacatggatg 11160ggctcaccct gggccagctc ggacggcttc tccagcacga caacgtcgtg aacatggatg 11160

cgcggaccgt actggcgagc gcaccggtga tgtatcccga gcaccaggcc ctctcgtccg 11220cgcggaccgt actggcgagc gcaccggtga tgtatcccga gcaccaggcc ctctcgtccg 11220

acgtcgagct gcactcctgg atcaccggta agcgggcact ccacgaggtc cgggcccgcc 11280acgtcgagct gcactcctgg atcaccggta agcgggcact ccacgaggtc cgggcccgcc 11280

gtatccccct gaccgaggtg gcgggctggg tccgggacga gtacaccatc caccgggagg 11340gtatccccct gaccgaggtg gcgggctggg tccgggacga gtacaccatc caccgggagg 11340

accagaggtt cttccaggtg ctggccgtcg ccgtggaggg agccaaccgg gaggtcccga 11400accagaggtt cttccaggtg ctggccgtcg ccgtggaggg agccaaccgg gaggtcccga 11400

gttggagtca gccgctgttc gaaccggtgg cccagggcgt cgtggccttc gcgtaccggg 11460gttggagtca gccgctgttc gaaccggtgg cccagggcgt cgtggccttc gcgtaccggg 11460

cgttcgccgg cgtaccgcac ctgctcgttc atgcccgggt cgaaggcgga ttcctcgaca 11520cgttcgccgg cgtaccgcac ctgctcgttc atgcccgggt cgaaggcgga ttcctcgaca 11520

ccatggaact cgcgccgacc gttcagtgcg tcccggcgaa ctacctccac ctgccggcct 11580ccatggaact cgcgccgacc gttcagtgcg tcccggcgaa ctacctccac ctgccggcct 11580

cgcaacgtcc gccgttcctc gacgtgatgc tggaggcacc ccaggaccgg atccggtacg 11640cgcaacgtcc gccgttcctc gacgtgatgc tggaggcacc ccaggaccgg atccggtacg 11640

ccgcgatcca ctccgaggag ggcggacggt tcctcaacgc cgagagccgc tacctcgtcg 11700ccgcgatcca ctccgaggag ggcggacggt tcctcaacgc cgagagccgc tacctcgtcg 11700

tcgccaccga cgagtcgacg acgccggccg agccgccgcc cggctaccac tggatcactc 11760tcgccaccga cgagtcgacg acgccggccg agccgccgcc cggctaccac tggatcactc 11760

cgggccagct cggatggctc gcaggtcaca gccgctacgt caacgtccag gcgaggacgc 11820cgggccagct cggatggctc gcaggtcaca gccgctacgt caacgtccag gcgaggacgc 11820

tgctcgccat cctcaccagt cgtgccgtcg acatcggtca gggctgttcg tgaacccgcc 11880tgctcgccat cctcaccagt cgtgccgtcg acatcggtca gggctgttcg tgaacccgcc 11880

cgcaacgggt cgtaccggtc ggtggggcag agccgcgact ggctctgccc caccgaccgg 11940cgcaacgggt cgtaccggtc ggtggggcag agccgcgact ggctctgccc caccgaccgg 11940

taccgccgtg tctcagcggg cggcgacagg acccttgccg tcgcccacct gccggatggt 12000taccgccgtg tctcagcggg cggcgacagg acccttgccg tcgcccacct gccggatggt 12000

ccggcgcatc gactccctgt cgttggcgcg cgggaagtcg tcgtccggca ccttcacccc 12060ccggcgcatc gactccctgt cgttggcgcg cgggaagtcg tcgtccggca ccttcacccc 12060

gaggaacgcg cacaggggct cccatccctg cttgacgtcg tagacgagaa ggttgtccgg 12120gaggaacgcg cacaggggct cccatccctg cttgacgtcg tagacgagaa ggttgtccgg 12120

ctcgacggag ttgacgacct cctcgttgtg cctgcggaag acctcgatgg cgtggtcctt 12180ctcgacggag ttgacgacct cctcgttgtg cctgcggaag acctcgatgg cgtggtcctt 12180

gtcggcgaac cgcccaccga acaggccgtc ccacaccatc gtgctcacga ggcggtgcac 12240gtcggcgaac cgcccaccga acaggccgtc ccacaccatc gtgctcacga ggcggtgcac 12240

ccgggccggc atggaaccgg ccggcgggcg gttctcactg ttccgcacgg cgaactggta 12300ccgggccggc atggaaccgg ccggcgggcg gttctcactg ttccgcacgg cgaactggta 12300

cagcgtgtcg tgggtgctgc ggtaccactc ctcgccgtca cggatggtga gaacgacctt 12360cagcgtgtcg tgggtgctgc ggtaccactc ctcgccgtca cggatggtga gaacgacctt 12360

ggcctccggg aaggcccgca cgatctgctt gtagtagatg gagctggggc cgtcgacagc 12420ggcctccggg aaggcccgca cgatctgctt gtagtagatg gagctggggc cgtcgacagc 12420

tgaggtgaag ccgtcgaaga gggccggcca gtcgggctgc tgcccgtcac agacgacctt 12480tgaggtgaag ccgtcgaaga gggccggcca gtcgggctgc tgcccgtcac agacgacctt 12480

ctcccactgg cccaggcggt gcggatcgct gacgacctcg aacatgtggt agcagggccc 12540ctcccactgg cccaggcggt gcggatcgct gacgacctcg aacatgtggt agcagggccc 12540

gtaaccgaga cgttccaacg cgaccttcag cgacgtggtg ccggtcctgc ccaacccggt 12600gtaaccgaga cgttccaacg cgaccttcag cgacgtggtg ccggtcctgc ccaacccggt 12600

gttgatgagc cgcaacaaag gagttccctt cttcagtgga tcgggtgccg tggctcagcg 12660gttgatgagc cgcaacaaag gagttccctt cttcagtgga tcgggtgccg tggctcagcg 12660

ccagtcaccc ggcgcctgcc gggtggacac gttgacgcag ttccacatat tgacgaggcc 12720ccagtcaccc ggcgcctgcc gggtggacac gttgacgcag ttccacatat tgacgaggcc 12720

gatgtgcagg atcagcgcac cgagttccgg ctcggtgaag tgccgggcgg cgtcgttcca 12780gatgtgcagg atcagcgcac cgagttccgg ctcggtgaag tgccgggcgg cgtcgttcca 12780

gacggtgtcc ggcaccgggt cctccttgtc gttgaggcgc gtcgcggcct cggcgagcgc 12840gacggtgtcc ggcaccgggt cctccttgtc gttgaggcgc gtcgcggcct cggcgagcgc 12840

cagggcggcg cgttcggatt cggtgaaaca gtcggcgtcg tgccaggcgt ccaccttgtg 12900cagggcggcg cgttcggatt cggtgaaaca gtcggcgtcg tgccaggcgt ccaccttgtg 12900

cagacgttga tcgatctggg ccgcagcgtc cggatcggtg ggcaggatgt tgacccggcc 12960cagacgttga tcgatctggg ccgcagcgtc cggatcggtg ggcaggatgt tgacccggcc 12960

gttgatctgg ctgacgcgca tgcggaccag atcgagcgtc cgcaccggca cgccgatctg 13020gttgatctgg ctgacgcgca tgcggaccag atcgagcgtc cgcaccggca cgccgatctg 13020

gttgatcgcc ttggcgaggt cgagcagcgg cagcagggcg tccggcacga ccagtgaagg 13080gttgatcgcc ttggcgaggt cgagcagcgg cagcagggcg tccggcacga ccagtgaagg 13080

gttgttcatc cgcggtgcca tggcattctc ctggggatgg gggcggctct ttcgggggaa 13140gttgttcatc cgcggtgcca tggcattctc ctggggatgg gggcggctct ttcgggggaa 13140

cggggagaga gatccggatc gcccggtgcg tcacggatgc gagacgcgag cggggagccg 13200cggggagaga gatccggatc gcccggtgcg tcacggatgc gagacgcgag cggggagccg 13200

gccttcacgc tctcgtgacg aggcggatcg gcccttgcgg gcggcgccga cgagcgcatg 13260gccttcacgc tctcgtgacg aggcggatcg gcccttgcgg gcggcgccga cgagcgcatg 13260

gtgcagccgt tcgtcgtcgt cctgcccggc ttcccactgc accccgagga cgaacggatg 13320gtgcagccgt tcgtcgtcgt cctgcccggc ttcccactgc accccgagga cgaacggatg 13320

cgctgccgcc tccaccacct cggcgacacc gtccggcgcc tcggccgtca ccgtgaggcc 13380cgctgccgcc tccaccacct cggcgacacc gtccggcgcc tcggccgtca ccgtgaggcc 13380

cgtgccgacc cggtcgattc cctgatggtg gtggcagcgg gcctcgggaa cgtcgttgcc 13440cgtgccgacc cggtcgattc cctgatggtg gtggcagcgg gcctcgggaa cgtcgttgcc 13440

cagcacgccg tcgatcgcgc tgccggcatg caggtccagg cgggtccggc cgagggtgaa 13500cagcacgccg tcgatcgcgc tgccggcatg caggtccagg cgggtccggc cgagggtgaa 13500

ggcctcggtc tcgggactgt gcccgtcgtg gccgacgagt tccggtaggt gctggtgcag 13560ggcctcggtc tcgggactgt gcccgtcgtg gccgacgagt tccggtaggt gctggtgcag 13560

cgtgcctccg tgcagcacgt tcagcagctg catgccgcgg cagatggcca ggacgggcag 13620cgtgcctccg tgcagcacgt tcagcagctg catgccgcgg cagatggcca ggacgggcag 13620

gccggcgtcg agcgcggcct cgatcagggc cagctcgacc cggtccgcct ccgggctgcc 13680gccggcgtcg agcgcggcct cgatcagggc cagctcgacc cggtccgcct ccgggctgcc 13680

gcaccgggtc cggggatgac gttcctgtcc gtacagcgcc ggatccacgt cggggccgcc 13740gcaccgggtc cggggatgac gttcctgtcc gtacagcgcc ggatccacgt cggggccgcc 13740

cggcaccagc agtccgtcga gctggggcac cgtgtgctcg gcgccgggca ccagcggcac 13800cggcaccagc agtccgtcga gctggggcac cgtgtgctcg gcgccgggca ccagcggcac 13800

cagcagcggg atgccgtccg ccgaggcgag catgtccacg tgggactgga gcgcgaggct 13860cagcagcggg atgccgtccg ccgaggcgag catgtccacg tgggactgga gcgcgaggct 13860

gacggttgtc tggaggccct gcacggtcac cggaaccgtt cgtgcgctga ttccgatcac 13920gacggttgtc tggaggccct gcacggtcac cggaaccgtt cgtgcgctga ttccgatcac 13920

gggacgcgcg gggccttccc gcctctgcgc ataattgttg aaggatgagt tcatggacac 13980gggacgcgcg gggccttccc gcctctgcgc ataattgttg aaggatgagt tcatggacac 13980

cctcataact tccggaaatc gccgttccag gcggttccgc cgcacgggac gatggtgcgg 14040cctcataact tccggaaatc gccgttccag gcggttccgc cgcacgggac gatggtgcgg 14040

cgaatgaggt ggggaatcat cggacgatgg cgggccggcg accatcgcgg ccgaccgtac 14100cgaatgaggt ggggaatcat cggacgatgg cgggccggcg accatcgcgg ccgaccgtac 14100

aacggttcct cggcgactcg caaccatcgc cggcggcttc gcgggaacgc tacggaatgg 14160aacggttcct cggcgactcg caaccatcgc cggcggcttc gcgggaacgc tacggaatgg 14160

gccgtcgtcc ggtggcgacc gagccggacc gccgccgtcg ccgatcgttc aactgaggcg 14220gccgtcgtcc ggtggcgacc gagccggacc gccgccgtcg ccgatcgttc aactgaggcg 14220

gagcgtgagc ggaaaccgct gttcagcggc gtgctcggct caccggggcg ccgtggcgtc 14280gagcgtgagc ggaaaccgct gttcagcggc gtgctcggct caccggggcg ccgtggcgtc 14280

catcggtcct tgaccgggac taaagaggct ttcgtcaaac tgcgaggcgt ccgtgcgggt 14340catcggtcct tgaccgggac taaagaggct ttcgtcaaac tgcgaggcgt ccgtgcgggt 14340

ccggcgtagc ggttcgatgt ccgtcgcgac accagcgcgt cacgaccggt acagtcatct 14400ccggcgtagc ggttcgatgt ccgtcgcgac accagcgcgt cacgaccggt acagtcatct 14400

tccggaagga cggcaatggc aacaatcgag gtcccggttc tcatcgtcgg cggcggtgga 14460tccggaagga cggcaatggc aacaatcgag gtcccggttc tcatcgtcgg cggcggtgga 14460

tgcggcttgt ccgcatctgt cttcctctcc gaccacggcg taggtcatct gctggtggaa 14520tgcggcttgt ccgcatctgt cttcctctcc gaccacggcg taggtcatct gctggtggaa 14520

cggcactcgg agacatcgag aatgccgaag gcgcattatc tgaaccagcg caccatgagc 14580cggcactcgg agacatcgag aatgccgaag gcgcattatc tgaaccagcg caccatgagc 14580

atcttccgtc agcacgggct cgacggcgag gtggccgaac tcggggcgtc gatcgaggag 14640atcttccgtc agcacgggct cgacggcgag gtggccgaac tcggggcgtc gatcgaggag 14640

ttcggaaagc tccggtggct gacgtcgctc ggcggcgacc ggccgctcga cggcatcgtc 14700ttcggaaagc tccggtggct gacgtcgctc ggcggcgacc ggccgctcga cggcatcgtc 14700

gtccaggaga tggacgcctt cggcggtggt gagctgcgcg agacctacga ggtggccggg 14760gtccaggaga tggacgcctt cggcggtggt gagctgcgcg agacctacga ggtggccggg 14760

ccggtcatgc cggtcaagct gccccaggtc cggctggagc cgatcctgcg ccggcacgcc 14820ccggtcatgc cggtcaagct gccccaggtc cggctggagc cgatcctgcg ccggcacgcc 14820

gagcaacgca atccgggccg gattctgttc agtcacgagc tggtcggctt cgccgaggag 14880gagcaacgca atccgggccg gattctgttc agtcacgagc tggtcggctt cgccgaggag 14880

ggcgaccggg tcgtggccga ggtccgcaac gtcgacaccg gtgaggtcac caccgtcgtg 14940ggcgaccggg tcgtggccga ggtccgcaac gtcgacaccg gtgaggtcac caccgtcgtg 14940

gcgcgctacc tcgtcgcggc cgacggcggc cgcacggtgg gcgccgcgct cggcgtacgg 15000gcgcgctacc tcgtcgcggc cgacggcggc cgcacggtgg gcgccgcgct cggcgtacgg 15000

atggaaggtc ttcccggttt tctccaggtc acgaccgcgt acttcaccgc ggacctgtcg 15060atggaaggtc ttcccggttt tctccaggtc acgaccgcgt acttcaccgc ggacctgtcg 15060

ccgtggtggc aggagggcac gctcctcacc cacttcctca acccctacga ccccgatctg 15120ccgtggtggc aggagggcac gctcctcacc cacttcctca acccctacga ccccgatctg 15120

tccagcaacc tcatcgagat gggaccgacc tggggcaagc actgcgagga gtgggtcctg 15180tccagcaacc tcatcgagat gggaccgacc tggggcaagc actgcgagga gtgggtcctg 15180

cacttcccgc cgggcgaccc ggagcgcttc ccgccggaga cgatcgtgcc caagatccgc 15240cacttcccgc cgggcgaccc ggagcgcttc ccgccggaga cgatcgtgcc caagatccgc 15240

gaggtcctcg ggctgcccga gctggaactc acgctgcacc acgtgacgaa ctgggccgtg 15300gaggtcctcg ggctgcccga gctggaactc acgctgcacc acgtgacgaa ctgggccgtg 15300

gagtccctgg tcgccgaccg ctaccgggtg ggccgggtgc tgctcgccgg cgatgccgtg 15360gagtccctgg tcgccgaccg ctaccgggtg ggccgggtgc tgctcgccgg cgatgccgtg 15360

caccggcagc cgcccaccgt cgggctgggg ctgaacaccg ggatccagga cgcgcacaac 15420caccggcagc cgcccaccgt cgggctgggg ctgaacaccg ggatccagga cgcgcacaac 15420

ctcgcgtgga agctcgccgc ggtgctcggc ggccgggcga acgacaggct gctggacacg 15480ctcgcgtgga agctcgccgc ggtgctcggc ggccgggcga acgacaggct gctggacacg 15480

tacgaggcgg agcggcaccc gatcgggcgg cacaacgtcg actgggcggt gtccgcggcc 15540tacgaggcgg agcggcaccc gatcgggcgg cacaacgtcg actgggcggt gtccgcggcc 15540

ctgcaccacc acgcgatcct cgaggccatc ggcctcggcc agcacacgcc gcagccgcgc 15600ctgcaccacc acgcgatcct cgaggccatc ggcctcggcc agcacacgcc gcagccgcgc 15600

cgcgaccggt cgttcgccgc gctcgtcgac ccgtctccgc tcgcagtcgc ggcgcgcgcc 15660cgcgaccggt cgttcgccgc gctcgtcgac ccgtctccgc tcgcagtcgc ggcgcgcgcc 15660

cggtcggcgg agatcttcgc cacccaccgg ggcgggtgcc aggcgcacga cgtggagatc 15720cggtcggcgg agatcttcgc cacccaccgg ggcgggtgcc aggcgcacga cgtggagatc 15720

ggcttcgcgt acgaggaggg agcggtcatc cccgacggca gtcagccgcc gccgcgggag 15780ggcttcgcgt acgaggaggg agcggtcatc cccgacggca gtcagccgcc gccgcgggag 15780

cccctgcggg acgtgtacca cccgacgacg cggcccggcc acgtgctgcc gcacgcgtgg 15840cccctgcggg acgtgtacca cccgacgacg cggcccggcc acgtgctgcc gcacgcgtgg 15840

atcgaaggtg gcggccgcgt cctgtccacc caggacctca ccggcacgtc gaccggcttc 15900atcgaaggtg gcggccgcgt cctgtccacc caggacctca ccggcacgtc gaccggcttc 15900

gtgctcatca ccggcccgca ggcggcaccc tggcaggagg cggccgcgga ggtcgccacc 15960gtgctcatca ccggcccgca ggcggcaccc tggcaggagg cggccgcgga ggtcgccacc 15960

aagttctccg tcccgatcgt caccgcgtcg atcggggccg gcggggagta cgtgggggcc 16020aagttctccg tcccgatcgt caccgcgtcg atcggggccg gcggggagta cgtgggggcc 16020

gacggccggt ggcaggacgt ccgggagatc acggacgagg gcgcgatcct ggtacggccc 16080gacggccggt ggcaggacgt ccgggagatc acggacgagg gcgcgatcct ggtacggccc 16080

gacaaccacg tcgcgtggcg cagcacgggc gccgcagcgg atcccgtcgg ggacctggcc 16140gacaaccacg tcgcgtggcg cagcacgggc gccgcagcgg atcccgtcgg ggacctggcc 16140

ggtgcgatcg ggcgcctcct cgacagccag ccggggccgg cgctcagctg acctctctct 16200ggtgcgatcg ggcgcctcct cgacagccag ccggggccgg cgctcagctg acctctctct 16200

ctgccacgcc cggaccatcg gtccgggcgt gggtcgttgt gccggccgac gtcagcgagc 16260ctgccacgcc cggaccatcg gtccgggcgt gggtcgttgt gccggccgac gtcagcgagc 16260

agtgcgcaca caaccgggcc ggggtggcaa ccgccacccc ggcccggccg gggacttcgg 16320agtgcgcaca caaccgggcc ggggtggcaa ccgccacccc ggcccggccg gggacttcgg 16320

aggaacgatc aggaggaggc cgctgtcgcg gcggtcctcg ggagcgcggc gagcacctgg 16380aggaacgatc aggaggaggc cgctgtcgcg gcggtcctcg ggagcgcggc gagcacctgg 16380

tcgacggcct tgcgtgcgtc ggcgatgcac gtgccgacgc cgaccccgtc gtacgcggcg 16440tcgacggcct tgcgtgcgtc ggcgatgcac gtgccgacgc cgaccccgtc gtacgcggcg 16440

ccacacacgg cgagcccgcg atgagcggcc agcgcggcac gcagccgggc cgtccggtcc 16500ccacacacgg cgagcccgcg atgagcggcc agcgcggcac gcagccgggc cgtccggtcc 16500

agatgaccga ctgtgtactg cggcagtgcg ctctcccacc tggtcacccg cgtcgccacc 16560agatgaccga ctgtgtactg cggcagtgcg ctctcccacc tggtcacccg cgtcgccacc 16560

ggggcaccca cggcgcccgt ggcctcggcc acctcggcgg cggccagcga cgcgaggtcg 16620ggggcaccca cggcgcccgt ggcctcggcc acctcggcgg cggccagcga cgcgaggtcg 16620

gcgtcgtcac gctgcagcag cgtctcgtcg ccgacccgcc cgatcgagca ccggaggatc 16680gcgtcgtcac gctgcagcag cgtctcgtcg ccgacccgcc cgatcgagca ccggaggatc 16680

tccacctcgc ccgccagatg cggccacttc accgtcgaga aggtcacctc gttcaccgcc 16740tccacctcgc ccgccagatg cggccacttc accgtcgaga aggtcacctc gttcaccgcc 16740

cgcccgtcca ccgccggcac gcggtacccg cagaacccgc gtgcggcgag accgccgggg 16800cgcccgtcca ccgccggcac gcggtacccg cagaacccgc gtgcggcgag accgccgggg 16800

aacgctccgc gcgggtacgc cagcgtgacg atcccgacgc tcgcgtacgg gatctccgcg 16860aacgctccgc gcgggtacgc cagcgtgacg atcccgacgc tcgcgtacgg gatctccgcg 16860

agctcggcgg cggccggggc gacaccgggt accccggcca gaagccggcc ggcgggcccg 16920agctcggcgg cggccggggc gacaccgggt accccggcca gaagccggcc ggcgggcccg 16920

gccggggcgg cgacgatcac cgcgtccgcg gtcagatgtt ccgtcccggc ggcggaggag 16980gccggggcgg cgacgatcac cgcgtccgcg gtcagatgtt ccgtcccggc ggcggaggag 16980

acggtgagcc gccagccgtg ttcgccgggg gagagggcgg tgacctccgc gccggtccgc 17040acggtgagcc gccagccgtg ttcgccgggg gagagggcgg tgacctccgc gccggtccgc 17040

aaggtcgctg ccgtcgagtc ggcgcgcacc gccgtggcca gcaccgtggc caggttcccg 17100aaggtcgctg ccgtcgagtc ggcgcgcacc gccgtggcca gcaccgtggc caggttcccg 17100

aggctgccgg tcagggtgcc gatgccggtc ggaggctcct gcccgtccgg gggcggcggc 17160aggctgccgg tcagggtgcc gatgccggtc ggaggctcct gcccgtccgg gggcggcggc 17160

gggatgagcg aggccgccgc cctggtcagt gacgggtgct tccgggacgc gttggccagg 17220gggatgagcg aggccgccgc cctggtcagt gacgggtgct tccgggacgc gttggccagg 17220

ggagccagca tggcctcgaa ggacaactcg tcggagcggc cggcgcacac gtcgctgagg 17280ggagccagca tggcctcgaa ggacaactcg tcggagcggc cggcgcacac gtcgctgagg 17280

aagggatcga cgagacggtc cagcacctcg cggccgagtc gctccgagac gtacgccgcg 17340aagggatcga cgagacggtc cagcacctcg cggccgagtc gctccgagac gtacgccgcg 17340

acggaaacgt cgccgttccg ctcggtcggg ggcagatcga cgtccctgcg ggcccgggcc 17400acggaaacgt cgccgttccg ctcggtcggg ggcagatcga cgtccctgcg ggcccgggcc 17400

acgccctccg cggagacgat gccggagcgg gcgagcgcgt ccatgtccga cggcacaccc 17460acgccctccg cggagacgat gccggagcgg gcgagcgcgt ccatgtccga cggcacaccc 17460

atgaactggc cggcgggcag cgaccggagt gcgcccctcg tccagatcgc cgtggcggtg 17520atgaactggc cggcgggcag cgaccggagt gcgcccctcg tccagatcgc cgtggcggtg 17520

gcgccggcca ggaccagctg gtcgccgagc ccggctgcgg tgagcaaccg ggttgtcttg 17580gcgccggcca ggaccagctg gtcgccgagc ccggctgcgg tgagcaaccg ggttgtcttg 17580

ggacgccgcg cgtacagccc ctcggccccc tcgtcgacag tgacaccggc cacctcggac 17640ggacgccgcg cgtacagccc ctcggccccc tcgtcgacag tgacaccggc cacctcggac 17640

gcggccagct tcccgccgag ccgcgacgac gcctcgagga gggtgatccg taccgatgcc 17700gcggccagct tcccgccgag ccgcgacgac gcctcgagga gggtgatccg taccgatgcc 17700

tcgcgaaggt agaaggctgc ggtcaaaccc gcgacgccgc ctcccacgat gaccacgtgt 17760tcgcgaaggt agaaggctgc ggtcaaaccc gcgacgccgc ctcccacgat gaccacgtgt 17760

ggcaccccgt ctgcccgttc cggttggttg tccaccatgt cgcctgctct ttcgtcggtg 17820ggcaccccgt ctgcccgttc cggttggttg tccaccatgt cgcctgctct ttcgtcggtg 17820

cgatgtccgt ggcagtttcg ccgaggcccg gtgaatcccg gtcgagcgcg ggtggacccg 17880cgatgtccgt ggcagtttcg ccgaggcccg gtgaatcccg gtcgagcgcg ggtggacccg 17880

gcggtgtgcg gtgtgaaccg cggccgttac tgtccggctt cctgcttccg cttcagctgc 17940gcggtgtgcg gtgtgaaccg cggccgttac tgtccggctt cctgcttccg cttcagctgc 17940

tcggcgaaca ggctccactc cttgctgatg ctgtccgcga tgggagcgat cacgctcagc 18000tcggcgaaca ggctccactc cttgctgatg ctgtccgcga tgggagcgat cacgctcagc 18000

cagtgcggcg ggatcgagcc ggcgacgtcg cgccaccgat ccggggcgcc cgcgtgccat 18060cagtgcggcg ggatcgagcc ggcgacgtcg cgccaccgat ccggggcgcc cgcgtgccat 18060

gccatccagc ggatgaacgc ccggccgccc tcggtgtagc ggatcgacgg gtcggtggcc 18120gccatccagc ggatgaacgc ccggccgccc tcggtgtagc ggatcgacgg gtcggtggcc 18120

agtttctccg ccacgccggc ccaggtgagt ggggcgtccg agcagccctt gcgtcgtacc 18180agtttctccg ccacgccggc ccaggtgagt ggggcgtccg agcagccctt gcgtcgtacc 18180

accgcgcggg gagccgccgg cgaggcggcg gtcgctgtcg cggctctgtc gtccggctcc 18240accgcgcggg gagccgccgg cgaggcggcg gtcgctgtcg cggctctgtc gtccggctcc 18240

ccgtccgggc ggtccgcggg ctggcggtgc ccgttgcgct cggggctgat gccacggcgg 18300ccgtccgggc ggtccgcggg ctggcggtgc ccgttgcgct cggggctgat gccacggcgg 18300

agccgggcgc tcacgtcgtg cacggtgccg agggagacgt ccgcctcccg ggcgacctgc 18360agccgggcgc tcacgtcgtg cacggtgccg agggagacgt ccgcctcccg ggcgacctgc 18360

cggagcgggg cgttcgggtg tgcgctgagg tactcggcgg cgcggcgccg gccctccgcg 18420cggagcgggg cgttcgggtg tgcgctgagg tactcggcgg cgcggcgccg gccctccgcg 18420

gcggtgaccg ggcgccgcct cccgtctcgg ccgatccgct tgctcccggg atcgacggcg 18480gcggtgaccg ggcgccgcct cccgtctcgg ccgatccgct tgctcccggg atcgacggcg 18480

cccgccgacc gggcgcgcag cgaggcgatg gtcttcgcgc tcagcccggt gatgccggcc 18540cccgccgacc gggcgcgcag cgaggcgatg gtcttcgcgc tcagcccggt gatgccggcc 18540

agggcgcgat cggaccagtc gggatgctgg gccaggaccc gatcggcgcc ggccaggcgg 18600agggcgcgat cggaccagtc gggatgctgg gccaggaccc gatcggcgcc ggccaggcgg 18600

tcggcccggg acaggggaag cccgtgggcg atgttggcct tcatcgccag gacgagtgcc 18660tcggcccggg acaggggaag cccgtgggcg atgttggcct tcatcgccag gacgagtgcc 18660

tcggaggcgg tgcagtccag gaagcgggcc ttgatgacct ggtcgccgcg gagccgggcg 18720tcggaggcgg tgcagtccag gaagcgggcc ttgatgacct ggtcgccgcg gagccgggcg 18720

gcttccagcc ggtgcaggcc gtcgatgacg caccagccgt cctcctggac gaggatcggc 18780gcttccagcc ggtgcaggcc gtcgatgacg caccagccgt cctcctggac gaggatcggc 18780

ggaagctccg cggtgccggc cgcgtcgacg agcagttgca cgtgggcggg gttcgtcccg 18840ggaagctccg cggtgccggc cgcgtcgacg agcagttgca cgtgggcggg gttcgtcccg 18840

ccctgacgca gacgaaggtt gggtgacagc gcgttgacgg gaacgtcccg cgccgggagc 18900ccctgacgca gacgaaggtt gggtgacagc gcgttgacgg gaacgtcccg cgccgggagc 18900

ctctcgaagt tcctcagatc gaatccgtcc ccgtcagtgg acccgtcgtc gtccgacacg 18960ctctcgaagt tcctcagatc gaatccgtcc ccgtcagtgg acccgtcgtc gtccgacacg 18960

gcgagcgcga ggctctccct agccagagca gcctgcccgt tgcggcgaga catccggtgc 19020gcgagcgcga ggctctccct agccagagca gcctgcccgt tgcggcgaga catccggtgc 19020

gacccccctc tgttgcggtt tgcgacgagg ttggcagagg gatctttagc cgcagtcgag 19080gacccccctc tgttgcggtt tgcgacgagg ttggcagagg gatctttagc cgcagtcgag 19080

acggggtgga cagggctagc cgtcggccgg tcgtaccccg gtgctggtga gggcgctgcc 19140acggggtgga cagggctagc cgtcggccgg tcgtaccccg gtgctggtga gggcgctgcc 19140

gaaccagtgg gtgagggtgt cggtgagggt gtcggtgctg ccgggtgagg tccaggcgac 19200gaaccagtgg gtgagggtgt cggtgagggt gtcggtgctg ccgggtgagg tccaggcgac 19200

gtagccgtcg ggtcggatga ggacggtgtc gagtggggtg tcgggttggt gggtccaggt 19260gtagccgtcg ggtcggatga ggacggtgtc gagtggggtg tcgggttggt gggtccaggt 19260

tccggtgacg gtgtcgacgc ggtcgtgcca gtggtgggtg ggtggtgtgg tggtgtcggt 19320tccggtgacg gtgtcgacgc ggtcgtgcca gtggtgggtg ggtggtgtgg tggtgtcggt 19320

ggtgacgagg acggcgcggg cggggtggag gaggtcggcg atgcgggtgt gggtgccgtc 19380ggtgacgagg acggcgcggg cggggtggag gaggtcggcg atgcgggtgt gggtgccgtc 19380

gggcagcgtg agttgatggt cgggtggcat gcgtcggccg agcagggggt ggtcgccggg 19440gggcagcgtg agttgatggt cgggtggcat gcgtcggccg agcagggggt ggtcgccggg 19440

gccggtgtcg tagcggatgc ccagcccgct gagcatcccg gcgagatgtc cggccgagtc 19500gccggtgtcg tagcggatgc ccagcccgct gagcatcccg gcgagatgtc cggccgagtc 19500

gcggtaggtg gccagctcgg cgagcacacc gcggaccggg tccatctcct cgccggtgag 19560gcggtaggtg gccagctcgg cgagcacacc gcggaccggg tccatctcct cgccggtgag 19560

ccgcagctcg atggcggccc gggcgttgcg ggccagctgc cgcccgatcg ggaggcgctc 19620ccgcagctcg atggcggccc gggcgttgcg ggccagctgc cgcccgatcg ggaggcgctc 19620

ggtctggtag gtgtcgagca ggccgtccgg cgcccagccc cgtaccgccg ccgccagctt 19680ggtctggtag gtgtcgagca ggccgtccgg cgcccagccc cgtaccgccg ccgccagctt 19680

ccaaccgagg ttcaccgcgt cctggacgcc cacgctcaaa ccctgcgcca tggccggtgg 19740ccaaccgagg ttcaccgcgt cctggacgcc cacgctcaaa ccctgcgcca tggccggtgg 19740

ctgcacgtga gcggcgtcgc cggccaggaa cacccggcca cgccggtact ccgcggcgac 19800ctgcacgtga gcggcgtcgc cggccaggaa cacccggcca cgccggtact ccgcggcgac 19800

gcgcgaggcg tcggtgaagc agctgatcca gcggcaccgt ccgtggtgga tggactcacc 19860gcgcgaggcg tcggtgaagc agctgatcca gcggcaccgt ccgtggtgga tggactcacc 19860

ggtgagccgc tgccacgcgt cggcgagttc ccggtagctc agcgagtcgc ggtccttcgg 19920ggtgagccgc tgccacgcgt cggcgagttc ccggtagctc agcgagtcgc ggtccttcgg 19920

gcgcatgccc cgctcgtgca cgacaacgcg ggtgacaccg ttctccaggt ccatcgccat 19980gcgcatgccc cgctcgtgca cgacaacgcg ggtgacaccg ttctccaggt ccatcgccat 19980

caccatgctg ccgccgggca ggttctcgcc gatgcgccgg cggcgggtgt cgatcccggt 20040caccatgctg ccgccgggca ggttctcgcc gatgcgccgg cggcgggtgt cgatcccggt 20040

gacgtcggcg gtgtagaacc cgcgggtggg ttccgggccg gtgaagtcga tgccggcgaa 20100gacgtcggcg gtgtagaacc cgcgggtggg ttccgggccg gtgaagtcga tgccggcgaa 20100

ccggcggacg gcgctgcgtc cgccgtcgca gcccaccagg taccgagcgg cgtcccggcc 20160ccggcggacg gcgctgcgtc cgccgtcgca gccaccagg taccgagcgg cgtcccggcc 20160

gctgccctcc gggccgtcga aggtgacgac gacgccgtcc gcgctctcct cgaagcccgt 20220gctgccctcc gggccgtcga aggtgacgac gacgccgtcc gcgctctcct cgaagcccgt 20220

gacctcgtat ccacggcgca ccggcacccc gagttcggcc acccagcctt ccagcatccg 20280gacctcgtat ccacggcgca ccggcacccc gagttcggcc acccagcctt ccagcatccg 20280

ctcggtgcgg aactgggaga ttccccggaa gccggcgtga tcctccggca gcaggccgag 20340ctcggtgcgg aactgggaga ttccccggaa gccggcgtga tcctccggca gcaggccgag 20340

gtcgaggggc acgccaccga agtggctgtc cgccggctcc gtcgcgccca gccggcccag 20400gtcgaggggc acgccaccga agtggctgtc cgccggctcc gtcgcgccca gccggcccag 20400

caacccacgc tggtcgaagc actcggccgt acgccgggtg aagctggcgc cgcgcgactc 20460caacccacgc tggtcgaagc actcggccgt acgccgggtg aagctggcgc cgcgcgactc 20460

cgtcggccgc tcgggcagct tctcgtagac gacgacatcg gctccgccga ggcgcagttc 20520cgtcggccgc tcgggcagct tctcgtagac gacgacatcg gctccgccga ggcgcagttc 20520

gccggccatc atcaggccga cggggccggc cccgaccacg atcacgtccc gggtcatcgc 20580gccggccatc atcaggccga cggggccggc cccgaccacg atcacgtccc gggtcatcgc 20580

gctgctccga tccggccggc gtcgacgctg ccgaaccagt gggtgagggt gtcggtgagg 20640gctgctccga tccggccggc gtcgacgctg ccgaaccagt gggtgagggt gtcggtgagg 20640

gtgtcggtgc tgccgggtga ggtccaggcg acgtagccgt cgggtcggat gaggacggtg 20700gtgtcggtgc tgccgggtga ggtccaggcg acgtagccgt cgggtcggat gaggacggtg 20700

tcgagtgggg tgtcgggttg gtgggtccag gttgcggtga cggtgtcgac gcggtcgtgc 20760tcgagtgggg tgtcgggttg gtgggtccag gttgcggtga cggtgtcgac gcggtcgtgc 20760

cagtggtggg tgggtggtgt ggtggtgtcg gtggtgacga ggacggcgcg ggcggggtgg 20820cagtggtggg tgggtggtgt ggtggtgtcg gtggtgacga ggacggcgcg ggcggggtgg 20820

aggaggtcgg cgatgcgggt gtgggtgccg tcgggcagcg tgagttgatg gtcgggtggc 20880aggaggtcgg cgatgcgggt gtgggtgccg tcgggcagcg tgagttgatg gtcgggtggc 20880

atgcgtcggc cgagcagggg gtggtcgccg gggccggtgt cgtagcggat gcccagcccg 20940atgcgtcggc cgagcagggg gtggtcgccg gggccggtgt cgtagcggat gcccagcccg 20940

ctgcccagac cgatgaggtg tgccgccgcc gccttcgcgg tggccaggtc ggacagcacg 21000ctgcccagac cgatgaggtg tgccgccgcc gccttcgcgg tggccaggtc ggacagcacg 21000

cgacgcaggg gttccacgcc gtcgtcgccg aggtacagca tcgacgcggc cttggcgttg 21060cgacgcaggg gttccacgcc gtcgtcgccg aggtacagca tcgacgcggc cttggcgttg 21060

ctgatcagct gccgtccgat cgggtggcgt tcggcctggt aggtgtcgag caggccgtcc 21120ctgatcagct gccgtccgat cgggtggcgt tcggcctggt aggtgtcgag caggccgtcc 21120

ggcgcccagc cccgtaccgt cgccgccagc ttccagccca ggttcaccgc gtccagcagg 21180ggcgcccagc cccgtaccgt cgccgccagc ttccagccca ggttcaccgc gtccagcagg 21180

ccggcgctga gcccccaggc ggcaagcggt ggcatctcgt gcgcggcgtc gccggcgagg 21240ccggcgctga gcccccaggc ggcaagcggt ggcatctcgt gcgcggcgtc gccggcgagg 21240

aagatccggc cgtgccggta ctcgtcggcc accgctgagg cgttgccgct ggcccacaac 21300aagatccggc cgtgccggta ctcgtcggcc accgctgagg cgttgccgct ggcccacaac 21300

cacggtgccg cgccgtggtg gatggattcg ccggtgagcc gctgccacgc gtcggcgacc 21360cacggtgccg cgccgtggtg gatggattcg ccggtgagcc gctgccacgc gtcggcgacc 21360

tcggtgaacg tcagtgccgc cggatcgggg tgcgggcgca gcgccctgtc gtggatcacg 21420tcggtgaacg tcagtgccgc cggatcgggg tgcgggcgca gcgccctgtc gtggatcacg 21420

atgcggtagc ggccctggcc gaccggagtg cacaccacca tgttgccgcc gggaaggcgt 21480atgcggtagc ggccctggcc gaccggagtg cacaccacca tgttgccgcc gggaaggcgt 21480

tcaccgatcg gtcgcggacg cagcgccacc ccggtgacct ccgcggtgta catgccccgg 21540tcaccgatcg gtcgcggacg cagcgccacc ccggtgacct ccgcggtgta catgccccgg 21540

gtggccggcg tcgcgtcggc cccgataccg gccagcgcac ggacggtgct ccccgcgccg 21600gtggccggcg tcgcgtcggc cccgataccg gccagcgcac ggacggtgct ccccgcgccg 21600

tcgcacccga cgaggtaggc ggcggtctcc tcgacgcgtc ccgcgggctc gtcgaaggtc 21660tcgcacccga cgaggtaggc ggcggtctcc tcgacgcgtc ccgcgggctc gtcgaaggtc 21660

gccacgaccc cgtgctccgt ctgccgcagg ccgaccagct cgtggcgccg gcggaccggt 21720gccacgaccc cgtgctccgt ctgccgcagg ccgaccagct cgtggcgccg gcggaccggt 21720

acgccgagtt cgttcagcca ccccgccagc atctcctcgg tccgggactg aggcaggccc 21780acgccgagtt cgttcagcca ccccgccagc atctcctcgg tccgggactg aggcaggccc 21780

agcacaccac tgtggtcgtc gtcgagcatg tcgagagcga tccgcacgcc tccgaagtgg 21840agcacaccac tgtggtcgtc gtcgagcatg tcgagagcga tccgcacgcc tccgaagtgg 21840

cccatcgggc cccaccggaa ggcgccgagc cgggggagca gaccgcgttg gtccagcgac 21900cccatcgggc cccaccggaa ggcgccgagc cgggggagca gaccgcgttg gtccagcgac 21900

tcggcggcgc gcctgttgaa gccgagcgcg cgggactcac cggtgggcgc ggcgagccgg 21960tcggcggcgc gcctgttgaa gccgagcgcg cgggactcac cggtgggcgc ggcgagccgg 21960

tcgtagacgc ggactctcgc cccacccagg tgcagttcgc cggcgagcat caggccgacc 22020tcgtagacgc ggactctcgc cccacccagg tgcagttcgc cggcgagcat caggccgacc 22020

gggcccgcgc ccaccacgat cacatcagtt tgcattgttt cggacctcca cgcgccgcgg 22080gggcccgcgc ccaccacgat cacatcagtt tgcattgttt cggacctcca cgcgccgcgg 22080

ccagagaatc gcgacgccaa agtacacatc atttccgcgc ggctcaacgt cgaaccatcg 22140ccagagaatc gcgacgccaa agtacacatc atttccgcgc ggctcaacgt cgaaccatcg 22140

aaacgcgctc aattgttcga ccgtgcattc ctcctggttg aaccggtggc cgcgcagagc 22200aaacgcgctc aattgttcga ccgtgcattc ctcctggttg aaccggtggc cgcgcagagc 22200

aatttcagag atgccggatg tcgtgctgcc gatcaggatg gacacaaaag cacgagggaa 22260aatttcagag atgccggatg tcgtgctgcc gatcaggatg gacacaaaag cacgagggaa 22260

ggattccaga cgtgccggtc ggcagaatcg cgctcatcac gggcgcgaac aagggcatcg 22320ggattccaga cgtgccggtc ggcagaatcg cgctcatcac gggcgcgaac aagggcatcg 22320

ggtacgagat cgcacgccag atgggggagc ggggatacgt cgtcctcgtc ggagcccggg 22380ggtacgagat cgcacgccag atgggggagc ggggatacgt cgtcctcgtc ggagcccggg 22380

acgaggtccg cggcaagcag gcggtcgagt cgctctcggc ccggggcgtc gaggccgtcc 22440acgaggtccg cggcaagcag gcggtcgagt cgctctcggc ccggggcgtc gaggccgtcc 22440

ggctgcggat cgacgtgacg gacgagacct ccgtcgtcga cgccgcggcc gagatcgagc 22500ggctgcggat cgacgtgacg gacgagacct ccgtcgtcga cgccgcggcc gagatcgagc 22500

ggcgctacgg ccgcctggac gtgctggtga acaacgccgg gatcgccggc ggctccaccg 22560ggcgctacgg ccgcctggac gtgctggtga acaacgccgg gatcgccggc ggctccaccg 22560

gtgcccccag cacggtgagc gcggccgatc tccgtcaggt gtacgagacg aacgtcctcg 22620gtgcccccag cacggtgagc gcggccgatc tccgtcaggt gtacgagacg aacgtcctcg 22620

gtgtcgtgtc ggtgacgaac gcgatgctcc cgctgctgcg ccgcgccgag gccgcgcgaa 22680gtgtcgtgtc ggtgacgaac gcgatgctcc cgctgctgcg ccgcgccgag gccgcgcgaa 22680

tcgtcaacat gtccagccac ctgggctccc tcaccctgaa ctcggcaccg gactcgccgt 22740tcgtcaacat gtccagccac ctgggctccc tcaccctgaa ctcggcaccg gactcgccgt 22740

tcgccggcct caacatggtc gcgtaccagt cgtcgaagac cgcgctgaac gccgtcactg 22800tcgccggcct caacatggtc gcgtaccagt cgtcgaagac cgcgctgaac gccgtcactg 22800

tcgcgtacgc caaggagctg cgggacacgc cgatcaaggt gaacgcggcg agccccggag 22860tcgcgtacgc caaggagctg cgggacacgc cgatcaaggt gaacgcggcg agccccggag 22860

tggtggcgac ggacctcaac caccaccgcg gaaaccggac tccggctcag ggggcggcga 22920tggtggcgac ggacctcaac caccaccgcg gaaaccggac tccggctcag ggggcggcga 22920

tcgcggtccg gctggcgacg ctcgacgaca cgggaccgtc gggcgcctgc ctggcggagg 22980tcgcggtccg gctggcgacg ctcgacgaca cgggaccgtc gggcgcctgc ctggcggagg 22980

aaggcgtcgt tccctggtga caggcccggg gcgacggatc gacgacttga cgacaagggg 23040aaggcgtcgt tccctggtga caggcccggg gcgacggatc gacgacttga cgacaagggg 23040

gacagggatc gtggcggtag atgggccgat tctggtgctg ggcggtaccg gccggcaggg 23100gacagggatc gtggcggtag atgggccgat tctggtgctg ggcggtaccg gccggcaggg 23100

tggtgcgacc gctcgcgagc tgctgaggcg agggcgggtg gtgcacgccc tggtccgcga 23160tggtgcgacc gctcgcgagc tgctgaggcg agggcgggtg gtgcacgccc tggtccgcga 23160

cccgcaggct cccgcggcgc gggcgctggc cgacgcgggc gccgtgctgg tgcacggcga 23220cccgcaggct cccgcggcgc gggcgctggc cgacgcgggc gccgtgctgg tgcacggcga 23220

cttcgacgac gaggcctcgt tgcgggcggc gatgaccggc gtgcacgggg tgttcagcgt 23280cttcgacgac gaggcctcgt tgcgggcggc gatgaccggc gtgcacgggg tgttcagcgt 23280

ccagacgttc cggacgcccg gcggggtggc caccgaggag cggcagggca aggcggtggc 23340ccagacgttc cggacgcccg gcggggtggc caccgaggag cggcagggca aggcggtggc 23340

cgacgcggcc gcccggaccg gcgtggcaca cctcgtctac agctccgtcg gcggcgcgga 23400cgacgcggcc gcccggaccg gcgtggcaca cctcgtctac agctccgtcg gcggcgcgga 23400

acggtccagc ggtgtcgacc acttcgagag caagtggcac gtcgagcagc acatccagaa 23460acggtccagc ggtgtcgacc acttcgagag caagtggcac gtcgagcagc acatccagaa 23460

gctcggtgtc ccggcgaccg tcctgcggcc gacgatgttc cacgaggtct tcctggacac 23520gctcggtgtc ccggcgaccg tcctgcggcc gacgatgttc cacgaggtct tcctggacac 23520

cggtccccgc ctcgtggacg gccggctggt cctcggcctg tggctgcggc ccgacgtccc 23580cggtccccgc ctcgtggacg gccggctggt cctcggcctg tggctgcggc ccgacgtccc 23580

gctgcagctc atcgcgacca gcgacatcgc cgggttcgcc gcggacgcct tcgaggatcc 23640gctgcagctc atcgcgacca gcgacatcgc cgggttcgcc gcggacgcct tcgaggatcc 23640

ggacacctgg ctgggccggc aggtggagat cgccggtgac gaactcaccg ggccgcagat 23700ggacacctgg ctgggccggc aggtggagat cgccggtgac gaactcaccg ggccgcagat 23700

ggccgaggcg ttcgcgcgcg tggcgggcat tccggcgcgc taccaggagc tgcccatcga 23760ggccgaggcg ttcgcgcgcg tggcgggcat tccggcgcgc taccaggagc tgcccatcga 23760

ccagctccga gccgttcgcc cggacctggc ggccatgttc gacttcttca acgacagggg 23820ccagctccga gccgttcgcc cggacctggc ggccatgttc gacttcttca acgacagggg 23820

ctaccgcgcc gacctgccgg cactccggcg catccggaag gacctggtca gcctggagga 23880ctaccgcgcc gacctgccgg cactccggcg catccggaag gacctggtca gcctggagga 23880

cttcctgcgt accagctgga ccgcccctgt ggcggcctga ccagccggcg cgacaagtgt 23940cttcctgcgt accagctgga ccgcccctgt ggcggcctga ccagccggcg cgacaagtgt 23940

gcccgccctc ccgatccggg gagggcgggc acatcgcgcg gatcaggtga cgaccgggcc 24000gcccgccctc ccgatccggg gagggcgggc acatcgcgcg gatcaggtga cgaccgggcc 24000

gctggtgcgg ctgcgcttga gctcgaagaa cccgtcggtc cgggccagcg cgagcacgcc 24060gctggtgcgg ctgcgcttga gctcgaagaa cccgtcggtc cgggccagcg cgagcacgcc 24060

gtcccagagc cggagcgcgg cgtcgccacg cgggaccggc gacaccaccg ggccgaagaa 24120gtcccagagc cggagcgcgg cgtcgccacg cgggaccggc gacaccaccg ggccgaagaa 24120

cgcgacgtcc cccaccgcga tgaccggtgt gcccacgtcg gtgccgaccc ggctgatgcc 24180cgcgacgtcc cccaccgcga tgaccggtgt gcccacgtcg gtgccgaccc ggctgatgcc 24180

gtcgtcgtgc gaggcccgaa cggcctcgtc gaactcgtcg gacccggccg ccgcggccac 24240gtcgtcgtgc gaggcccgaa cggcctcgtc gaactcgtcg gacccggccg ccgcggccac 24240

cccgggatcc atgccggcgg cgaccagcgc cgcctcgtag gtggcccggt cccgggacgc 24300cccgggatcc atgccggcgg cgaccagcgc cgcctcgtag gtggcccggt cccgggacgc 24300

ccggtcgacg tggaaccgcg tacccagttc ggtgtagagc cgtcccaggg aatccggccc 24360ccggtcgacg tggaaccgcg tacccagttc ggtgtagagc cgtcccaggg aatccggccc 24360

gtaccgctgc tcggcggcga cgcagacccg caccgcctcc agcgaggacc gcagacccgt 24420gtaccgctgc tcggcggcga cgcagacccg caccgcctcc agcgaggacc gcagacccgt 24420

ccggtaccgc tcgggcaggt cgtcgcggcc ctcgttgagc accgccagac tcatgacgtg 24480ccggtaccgc tcgggcaggt cgtcgcggcc ctcgttgagc accgccagac tcatgacgtg 24480

ccagcacggc ctgaccggac ggtgttcggc gacttcgaga atccaccgag aggtgatcca 24540ccagcacggc ctgaccggac ggtgttcggc gacttcgaga atccaccgag aggtgatcca 24540

cgcccatgga cacgtcgggt cgaaccagaa gtccacagtg gtaccgacac cgtccggcat 24600cgcccatgga cacgtcgggt cgaaccagaa gtccacagtg gtaccgacac cgtccggcat 24600

cgccggcctc ctcgcagtgc ttgtgatcgg aactcgtacg actcgcgagt gcggccggcg 24660cgccggcctc ctcgcagtgc ttgtgatcgg aactcgtacg actcgcgagt gcggccggcg 24660

gatcagccga gcggctccca gtggtagaag cgggtggcca tggcgtccgc gggggagcgc 24720gatcagccga gcggctccca gtggtagaag cgggtggcca tggcgtccgc gggggagcgc 24720

caggtcgccg ggtcgtacgg ctcgatgaac ggcttcaggt cctcgctgat cctgatgaac 24780caggtcgccg ggtcgtacgg ctcgatgaac ggcttcaggt cctcgctgat cctgatgaac 24780

cgggggtcgg tgcgggcctc gatgatccgc tcgccgccgt actcgccgtc gaagtcctgc 24840cgggggtcgg tgcgggcctc gatgatccgc tcgccgccgt actcgccgtc gaagtcctgc 24840

aggtggaagt acaaccctcg atagctgaag agctcacgac gccgggtgcc catgcgctgc 24900aggtggaagt acaaccctcg atagctgaag agctcacgac gccgggtgcc catgcgctgc 24900

ggcatgtcgg tccggtcgaa cgcgccgaac agttcggcca cctcggtcga cgactgcggt 24960ggcatgtcgg tccggtcgaa cgcgccgaac agttcggcca cctcggtcga cgactgcggt 24960

tgcatccgac cgacgatcag ggtgctgtgc atgtcaagct caccgccttc gttgcgggat 25020tgcatccgac cgacgatcag ggtgctgtgc atgtcaagct caccgccttc gttgcgggat 25020

cgggggtgga cgcgatcagt agttcccgag gccgccgcag acgttcatcg cctgcgcggt 25080cgggggtgga cgcgatcagt agttcccgag gccgccgcag acgttcatcg cctgcgcggt 25080

caccggggcg gcggcatcac tgaccagata cccgacgagg ccggcgacct cctcgggggt 25140caccggggcg gcggcatcac tgaccagata cccgacgagg ccggcgacct cctcgggggt 25140

ggagtaccgg ccgaggggga tcttcgcctc gaaccgctcg aggatcgccg cctcggtcgt 25200ggagtaccgg ccgaggggga tcttcgcctc gaaccgctcg aggatcgccg cctcggtcgt 25200

ctcgaagtgc gcggcgtagc cctgccgtac ccgctgggcc atgggcgtct cgacgtagcc 25260ctcgaagtgc gcggcgtagc cctgccgtac ccgctgggcc atgggcgtct cgacgtagcc 25260

gggacagacg gcgttcaccg tgaccccggt cttcgccagt tccagtccca gcgccttcgt 25320gggacagacg gcgttcaccg tgaccccggt cttcgccagt tccagtccca gcgccttcgt 25320

gaagccgacg acaccgtgct tggacgcgga gtacggggcg cccagcacca caccctgctt 25380gaagccgacg acaccgtgct tggacgcgga gtacggggcg cccagcacca caccctgctt 25380

gccgccggtc gacgcgatgt tgatgatccg tccccagccg cggtcgagcg acccgcccga 25440gccgccggtc gacgcgatgt tgatgatccg tccccagccg cggtcgagcg acccgcccga 25440

ccggagcacc tcccgggtca cccggaagac gccgttcagg ttggtctcga tgacgtcgta 25500ccggagcacc tcccgggtca cccggaagac gccgttcagg ttggtctcga tgacgtcgta 25500

ccagagttcg tcactgatgt cgctggtcac cccgccgccg ctgcggccgg cgctgttgac 25560ccagagttcg tcactgatgt cgctggtcac cccgccgccg ctgcggccgg cgctgttgac 25560

caggacgtcg acacgcccgt acctgcgcac ggcggcagcc accagctgtt cgacgtcctc 25620caggacgtcg acacgcccgt acctgcgcac ggcggcagcc accagctgtt cgacgtcctc 25620

accgcgccgg acgtcggcgg tcacgccgtc ggcgtcgtga ccggccgcac gtagcttgcc 25680accgcgccgg acgtcggcgg tcacgccgtc ggcgtcgtga ccggccgcac gtagcttgcc 25680

gaccgtctcc tcgacggatt cggggttccg cgcacagacg aagaccttga ggccgtcacg 25740gaccgtctcc tcgacggatt cggggttccg cgcacagacg aagaccttga ggccgtcacg 25740

ggcgagtctt tccgccaccg caaggccgat tccgctggtt ccaccggtga tcagcgcaac 25800ggcgagtctt tccgccaccg caaggccgat tccgctggtt ccaccggtga tcagcgcaac 25800

tcgctgtccg ttgtccgtca ttcgacccgc ccctcaccaa tatcgtgtcg cggcagtatc 25860tcgctgtccg ttgtccgtca ttcgacccgc ccctcaccaa tatcgtgtcg cggcagtatc 25860

tcagcaattt cagaaatctc actcgagcct cggtggaagc atttgtttca accgctccgg 25920tcagcaattt cagaaatctc actcgagcct cggtggaagc atttgtttca accgctccgg 25920

ctctccagtt gaacgacctc ggcggagctt gcgatcggaa aaggcaccga tctatcgtct 25980ctctccagtt gaacgacctc ggcggagctt gcgatcggaa aaggcaccga tctatcgtct 25980

ggcaaccccg caatttccca ttcctggagg agaatcatgt ccgcattgcc taccgtgccc 26040ggcaaccccg caatttccca ttcctggagg agaatcatgt ccgcattgcc taccgtgccc 26040

gcagtgcggg acaactcggc ggcctacgcg ctgctcgatc tgatccaggg ttcggtgatc 26100gcagtgcggg acaactcggc ggcctacgcg ctgctcgatc tgatccaggg ttcggtgatc 26100

acccaggcga tctccgtggc ggccaagctc ggcgtcgccg acgttctcgc cgacgggccc 26160acccaggcga tctccgtggc ggccaagctc ggcgtcgccg acgttctcgc cgacgggccc 26160

ctgccggccg aggagatcgc caagcgcgtc ggatccgaca gcgaggcgac gtaccggctg 26220ctgccggccg aggagatcgc caagcgcgtc ggatccgaca gcgaggcgac gtaccggctg 26220

ctgaggactc tctccggttg ctcggtgttc gcgcttcggg ccgacggccg gttcgagctc 26280ctgaggactc tctccggttg ctcggtgttc gcgcttcggg ccgacggccg gttcgagctc 26280

acgccgatgg gtgacgcgct gcgcgagggc gccccggact cgatgcgcgg catcgccatg 26340acgccgatgg gtgacgcgct gcgcgagggc gccccggact cgatgcgcgg catcgccatg 26340

ctgatggggc acccgctgct ctgggaggag tgggcgcacc tcatcgagtc ggtccgcacc 26400ctgatggggc acccgctgct ctgggaggag tgggcgcacc tcatcgagtc ggtccgcacc 26400

ggcgaggcca acatgcccaa gctgcgcggc atgggcgcct acgagttcct catgtcgaac 26460ggcgaggcca acatgcccaa gctgcgcggc atgggcgcct acgagttcct catgtcgaac 26460

cccgcgtacg ccgccgagtt cttccagggc atgggcagca tgtccgactc ggagaccgac 26520cccgcgtacg ccgccgagtt cttccagggc atgggcagca tgtccgactc ggagaccgac 26520

ccggtcctcg cggcgtacga cttctcgagc ttcggcacca tcgtggacgt cgtcggcggg 26580ccggtcctcg cggcgtacga cttctcgagc ttcggcacca tcgtggacgt cgtcggcggg 26580

cgcggccggc tcctggccgg catcctcgcc ggggccacca aggcccgggg catcctgtac 26640cgcggccggc tcctggccgg catcctcgcc ggggccacca aggcccgggg catcctgtac 26640

gacaacgagg tcgcgaccgc tgacgcgccg gcgacgctcg aggccgcagg cgtcgccgac 26700gacaacgagg tcgcgaccgc tgacgcgccg gcgacgctcg aggccgcagg cgtcgccgac 26700

cgggtcacga tcgagaacgg ttcccacttc gacaagctgc cggccggggc cgacgcctac 26760cgggtcacga tcgagaacgg ttcccacttc gacaagctgc cggccggggc cgacgcctac 26760

gtgctgaagc acatcctgca cgacttcccg gagcaggcat gcctgcagct gctgcagaac 26820gtgctgaagc acatcctgca cgacttcccg gagcaggcat gcctgcagct gctgcagaac 26820

gtccgggagg cgatcgctcc cggcggccgg atgctcgtga tcgagtacgt gctcgaggag 26880gtccgggagg cgatcgctcc cggcggccgg atgctcgtga tcgagtacgt gctcgaggag 26880

aacaacaagc gccacatcgg aaacatcatc gacctgtggc tgctgctcct cctcggcgcg 26940aacaacaagc gccacatcgg aaacatcatc gacctgtggc tgctgctcct cctcggcgcg 26940

aaggagcgca ccctgccgca gtacaccgaa ctcttcgcca aggcgggcat gaaggtcacc 27000aaggagcgca ccctgccgca gtacaccgaa ctcttcgcca aggcgggcat gaaggtcacc 27000

agggtgatcc ccaccgcctc gccggtctcg atcatcgaag ccactcccgc ctgagccgcg 27060agggtgatcc ccaccgcctc gccggtctcg atcatcgaag ccactcccgc ctgagccgcg 27060

gcgggcacgc cccggctgcg ggactcccac gagaagcgag gtcagatcat ggatacggat 27120gcgggcacgc cccggctgcg ggactcccac gagaagcgag gtcagatcat ggatacggat 27120

gtcatcgtca tcggtgctgg tcccacggga ctgatgctcg ctgccgaact tcgcctcggt 27180gtcatcgtca tcggtgctgg tcccacggga ctgatgctcg ctgccgaact tcgcctcggt 27180

ggcgcggagg tcaccgtctt cgaacggcgg agcgagcgga gctgggagtc acgcggcatc 27240ggcgcggagg tcaccgtctt cgaacggcgg agcgagcgga gctgggagtc acgcggcatc 27240

ggcttcacgg cgcgcgcggc ggaggtgttc cagcagcgcg gcctcctcga gaggctggag 27300ggcttcacgg cgcgcgcggc ggaggtgttc cagcagcgcg gcctcctcga gaggctggag 27300

aacaccgaga tcacccggca ggggcacttc ggcggcatcc cgctcgactt cggggtgctc 27360aacaccgaga tcacccggca ggggcacttc ggcggcatcc cgctcgactt cggggtgctc 27360

gaggactcgc acttcggcgt gcgcggcgca ccccagttcg tggtggagga gatgctggag 27420gaggactcgc acttcggcgt gcgcggcgca ccccagttcg tggtggagga gatgctggag 27420

aagcgggcgc tggaactcgg tgtgactctg caccgcgggc acgagctcac cgacctggcc 27480aagcgggcgc tggaactcgg tgtgactctg caccgcgggc acgagctcac cgacctggcc 27480

gattcgggag acggcgtcac ggtgaccgtg cacggtcccg acggccgtgc cgagtactcg 27540gattcgggag acggcgtcac ggtgaccgtg cacggtcccg acggccgtgc cgagtactcg 27540

gcccggtacc tcgtcgggtg tgacggcggc cggagcacgg tgcggaagct ggccgccttc 27600gcccggtacc tcgtcgggtg tgacggcggc cggagcacgg tgcggaagct ggccgccttc 27600

gacttcccgg gccgggacgc gacgtgcgag atgtacctgg cagacgtcac cggctgcgac 27660gacttcccgg gccgggacgc gacgtgcgag atgtacctgg cagacgtcac cggctgcgac 27660

atccggcccc ggatgatcgg cgagctgctc ccgaacggca tggtcatggc gggcccgctc 27720atccggcccc ggatgatcgg cgagctgctc ccgaacggca tggtcatggc gggcccgctc 27720

ggcgagggct acttccgcat catcgtctgc gagaccggta cgccgccgga ccgcaaccgc 27780ggcgagggct acttccgcat catcgtctgc gagaccggta cgccgccgga ccgcaaccgc 27780

caggtcacct tcgccgacgt ggcggacgcc tggcagcggc tgaccggcga ctccatccac 27840caggtcacct tcgccgacgt ggcggacgcc tggcagcggc tgaccggcga ctccatccac 27840

ggcggcgagg cgcgatgggt cagccgcttc accgacgcca cgcggcaggt caccgagtac 27900ggcggcgagg cgcgatgggt cagccgcttc accgacgcca cgcggcaggt caccgagtac 27900

cggcggggcc gcgtcctgct ggccggcgac gccgcgcaca tccacctccc cgccggcggc 27960cggcggggcc gcgtcctgct ggccggcgac gccgcgcaca tccacctccc cgccggcggc 27960

caggggatga gcatcggcct gcaggacgcg gtgaacctcg gctggaagct cgcggcggcg 28020caggggatga gcatcggcct gcaggacgcg gtgaacctcg gctggaagct cgcggcggcg 28020

gtccggggga cgggggacga cgcccttctc gacacgtatc acagcgagcg gcacccggtg 28080gtccggggga cgggggacga cgcccttctc gacacgtatc acagcgagcg gcacccggtg 28080

ggcgcccggg tcctgcgcaa cacgcgggcc caggggaccc tgaacctcag cggcaaggcg 28140ggcgcccggg tcctgcgcaa cacgcgggcc caggggaccc tgaacctcag cggcaaggcg 28140

acggagccgc tgcgggccgt cgtggcggag ctcatcgcgc tgcccgtggt ggcgcgacac 28200acggagccgc tgcgggccgt cgtggcggag ctcatcgcgc tgcccgtggt ggcgcgacac 28200

ctgtccggca tggtgagcgg actcgacatc cggtacgacg tcggcgcgga gggccacccg 28260ctgtccggca tggtgagcgg actcgacatc cggtacgacg tcggcgcgga gggccacccg 28260

ctcctcggcg cgcggatccc ggaccgggac gtggacctcg cggacggcgg caccgaccgg 28320ctcctcggcg cgcggatccc ggaccgggac gtggacctcg cggacggcgg caccgaccgg 28320

atcgcgcggc tgctgcacac cgcgcgcggc gtgctgatca ccgccgacgg ctccggggaa 28380atcgcgcggc tgctgcacac cgcgcgcggc gtgctgatca ccgccgacgg ctccggggaa 28380

accagccgtc gcgccgcgcc ctggtccgac cgggtcgacg tggtacgcgt ccgcagcgtg 28440accagccgtc gcgccgcgcc ctggtccgac cgggtcgacg tggtacgcgt ccgcagcgtg 28440

ccggtcgggt cgcgcgagga cggggccgcg ccggaatcgg tgctggtccg cccggacggg 28500ccggtcgggt cgcgcgagga cggggccgcg ccggaatcgg tgctggtccg cccggacggg 28500

cacgtcgcct gggtcgcgcc ggacggaggc gacctcgacg aggcgctgcg gcgctggttc 28560cacgtcgcct gggtcgcgcc ggacggaggc gacctcgacg aggcgctgcg gcgctggttc 28560

ggcgcgcccc ggccggccgg cgacacggcg gactccccgc agctcgcgtc cgcccgatag 28620ggcgcgcccc ggccggccgg cgacacggcg gactccccgc agctcgcgtc cgcccgatag 28620

acgtcccgtc gccagaggag gcaaacgtga ccaccgatac ggtccacgag acggagcacc 28680acgtcccgtc gccagaggag gcaaacgtga ccaccgatac ggtccacgag acggagcacc 28680

tgatcgtcgt cgacgctccc gccgagcggg tctacgccct gatcgaggat gtcggcacct 28740tgatcgtcgt cgacgctccc gccgagcggg tctacgccct gatcgaggat gtcggcacct 28740

ggccggaggt gttcccgccg accgtgcacg ccgagtgtct cgaacgcgac ggcgacaccg 28800ggccggaggt gttcccgccg accgtgcacg ccgagtgtct cgaacgcgac ggcgacaccg 28800

agctgatcag gatctgggcg accgccaacg gcgtcgccaa gacgtggacg tcccgccgcc 28860agctgatcag gatctgggcg accgccaacg gcgtcgccaa gacgtggacg tcccgccgcc 28860

ggcacgaccc cgggcgcctg agcgtgtcgt tccggcagga gcggtcgcag catccggtcg 28920ggcacgaccc cgggcgcctg agcgtgtcgt tccggcagga gcggtcgcag catccggtcg 28920

gcggcatggg cggggcgtgg gtgatcgagc cgatcaccga cgccacctgc cgggtacggc 28980gcggcatggg cggggcgtgg gtgatcgagc cgatcaccga cgccacctgc cgggtacggc 28980

tcctgcacga cttcttcccg gccagcgacg agcccgccga cctcgagtgg atcaagcagg 29040tcctgcacga cttcttcccg gccagcgacg agcccgccga cctcgagtgg atcaagcagg 29040

cggtggaccg caacagcgcg tcggagctgc aggcgctgaa gtccagcgcc gagccggcgg 29100cggtggaccg caacagcgcg tcggagctgc aggcgctgaa gtccagcgcc gagccggcgg 29100

agccgggcca gtgcttcacc ttcgccgaca ccgtgacggt cgagggcagc gccgaggacg 29160agccgggcca gtgcttcacc ttcgccgaca ccgtgacggt cgagggcagc gccgaggacg 29160

tctacgactt cctcaacaac gcccagctgt ggccgcagcg gctcccgcac gtcgcccggg 29220tctacgactt cctcaacaac gcccagctgt ggccgcagcg gctcccgcac gtcgcccggg 29220

tctcgctcga ggaggattcg ccgggcctcc aggtgctcga gatggacacc cggaccaagg 29280tctcgctcga ggaggattcg ccgggcctcc aggtgctcga gatggacacc cggaccaagg 29280

acgggtcggt gcacacgacg cgctccgtcc gcgtgtgtga accccaccgc agcatcgtgt 29340acgggtcggt gcacacgacg cgctccgtcc gcgtgtgtga accccaccgc agcatcgtgt 29340

acaagcagac cgtcacgccc gcactgatga cgctgcacac cggccgctgg ctcatcgagc 29400acaagcagac cgtcacgccc gcactgatga cgctgcacac cggccgctgg ctcatcgagc 29400

cgcagggcgc cggccaggtg gcggtcacct cgcggcacac ggtccggatc aacaccgcgc 29460cgcagggcgc cggccaggtg gcggtcacct cgcggcacac ggtccggatc aacaccgcgc 29460

ggatcaccga gctcctgggc ccggaggccg atctggccac ggcgcagcag ctggtccgca 29520ggatcaccga gctcctgggc ccggaggccg atctggccac ggcgcagcag ctggtccgca 29520

acgccctcag tgccaacagc ctgacgaccc tgcgggccgc gaaggcgtac gccgagggcg 29580acgccctcag tgccaacagc ctgacgaccc tgcgggccgc gaaggcgtac gccgagggcg 29580

gcggcggcag gcacccggtc ccgtgaccgt ctccccggtc gtcgtcgtcg gtgctggccc 29640gcggcggcag gcacccggtc ccgtgaccgt ctccccggtc gtcgtcgtcg gtgctggccc 29640

ggtcggactc atgctggcct gcgaactcgg acgggctcgc gtgccggtcg tcgtcgtgga 29700ggtcggactc atgctggcct gcgaactcgg acgggctcgc gtgccggtcg tcgtcgtgga 29700

gcgtctcgcc acgccgatga ccgagtcgcg ggccagccag ctctccacgt tgaccgcgga 29760gcgtctcgcc acgccgatga ccgagtcgcg ggccagccag ctctccacgt tgaccgcgga 29760

gttgctgcac gagcgcgggt tcgacgagct gctcgacgag gcggtgcacg agccgcgcgc 29820gttgctgcac gagcgcgggt tcgacgagct gctcgacgag gcggtgcacg agccgcgcgc 29820

gcacttcgcg ggtctcgcgt tcgacctgtc gcagctggac agcgactacc cgggcggctg 29880gcacttcgcg ggtctcgcgt tcgacctgtc gcagctggac agcgactacc cgggcggctg 29880

gaaggtgccc cagtaccgga ccgaggccgt cctcgggcgg caggccgaac gcctcggcgt 29940gaaggtgccc cagtaccgga ccgaggccgt cctcgggcgg caggccgaac gcctcggcgt 29940

gacagtgctg cgggcgcagg agctgaccgg gctcaccgag cggccggacc acgtggtgtg 30000gacagtgctg cgggcgcagg agctgaccgg gctcaccgag cggccggacc acgtggtgtg 30000

ccggctccgg ggaccagacg gtgatcggac cgtcagggcc cgcttcgtgg tcgggtgcga 30060ccggctccgg ggaccagacg gtgatcggac cgtcagggcc cgcttcgtgg tcgggtgcga 30060

cggggcgcac agcaccgtac gccggctgca cgggttcccg gcgtcggtca cgccggcgac 30120cggggcgcac agcaccgtac gccggctgca cgggttcccg gcgtcggtca cgccggcgac 30120

gaaggagctg ctgcgggcgg acgtgacggg tgtccggatc cgcgaccgca ggttcgagcg 30180gaaggagctg ctgcgggcgg acgtgacggg tgtccggatc cgcgaccgca ggttcgagcg 30180

gctggacggc gggttcgcgg tggcggcgac ccgcgacggt gtgacacggg tgatggtgca 30240gctggacggc gggttcgcgg tggcggcgac ccgcgacggt gtgacacggg tgatggtgca 30240

cccgcgcggc cggccggtga cccggcgcag cggtcctccc gatttcggcg aggtgatccg 30300cccgcgcggc cggccggtga cccggcgcag cggtcctccc gatttcggcg aggtgatccg 30300

ggcctggcgg gacgtcaccg gcgaggacct gtccggcggg accgccgtct ggctcgacgc 30360ggcctggcgg gacgtcaccg gcgaggacct gtccggcggg accgccgtct ggctcgacgc 30360

gttcgacaac tccagaggcc aggccgacgc gtaccggcgt ggccgggtcc tgctggccgg 30420gttcgacaac tccagaggcc aggccgacgc gtaccggcgt ggccgggtcc tgctggccgg 30420

cgacgccgcc cactggcaca tgccgatcgg cggtcaggct ctcaacgtcg ggctccagga 30480cgacgccgcc cactggcaca tgccgatcgg cggtcaggct ctcaacgtcg ggctccagga 30480

cgccgtcaac ctgggctgga agctggccgc gacagtggac ggccgggccg ctccggggct 30540cgccgtcaac ctgggctgga agctggccgc gacagtggac ggccgggccg ctccggggct 30540

gctggacagc taccacgacg agcggcaccc cgtcgcggcg cgcgtgctcg accacgtggc 30600gctggacagc taccacgacg agcggcaccc cgtcgcggcg cgcgtgctcg accacgtggc 30600

ggcgcaggag atgttgctgc tgggcggcgc cgagatcgat ccgctgcgcg cggtcctcgc 30660ggcgcaggag atgttgctgc tgggcggcgc cgagatcgat ccgctgcgcg cggtcctcgc 30660

cgaactcgtc gcactcggcc aggtccgcgc ccacctggcg gagacggccg ccaacgtcgg 30720cgaactcgtc gcactcggcc aggtccgcgc ccacctggcg gagacggccg ccaacgtcgg 30720

cgaccgttac ggcccgccga catcgccgct ggtggggcgg cgggtggtga acctgcggct 30780cgaccgttac ggcccgccga catcgccgct ggtggggcgg cgggtggtga acctgcggct 30780

gcgcacggac tcgggaccgc ttccggtggc gaaccccggg ccggtcgtgg tccggctcgc 30840gcgcacggac tcgggaccgc ttccggtggc gaaccccggg ccggtcgtgg tccggctcgc 30840

ccccggcccg gacgccgtcc gccgcaccgt tcccacggtc cacgcacgtc ccgacaaggg 30900ccccggcccg gacgccgtcc gccgcaccgt tcccacggtc cacgcacgtc ccgacaaggg 30900

ttcccttccg ggaaccactg ccctcctcct gcggcccgac ggctacatcg cgtgggccgg 30960ttcccttccg ggaaccactg ccctcctcct gcggcccgac ggctacatcg cgtgggccgg 30960

cgacgacgag gacggcctga accgggcgat cgacaagctg ctactcgcgg aaggtaggac 31020cgacgacgag gacggcctga accgggcgat cgacaagctg ctactcgcgg aaggtaggac 31020

ggacaatgcc agagtttgac gcggacgtca tcatcgtggg ggccggcccc accgggctca 31080ggacaatgcc agagtttgac gcggacgtca tcatcgtggg ggccggcccc accgggctca 31080

tgctcgccgg ggagcttcgc ctggccggcg tcgaggccct cgtcctggac cggctcgccg 31140tgctcgccgg ggagcttcgc ctggccggcg tcgaggccct cgtcctggac cggctcgccg 31140

agccgacgaa gcagtcccgg gcgctcggct tctcggcacg caccatcgag gagttcgacc 31200agccgacgaa gcagtcccgg gcgctcggct tctcggcacg caccatcgag gagttcgacc 31200

agcgcgggct cctgccccgc ttcggcgagc tgcagaccat ccccttcggt cacttcggcg 31260agcgcgggct cctgccccgc ttcggcgagc tgcagaccat ccccttcggt cacttcggcg 31260

ggctcccgct ggactaccgc gtcgtcgagg gcggctcgta cggcgctcgg ggcaagccgc 31320ggctcccgct ggactaccgc gtcgtcgagg gcggctcgta cggcgctcgg ggcaagccgc 31320

agtccctcac cgagggcgtg ctgaccggct gggccgccga gcagggcgcc gccgtcctcc 31380agtccctcac cgagggcgtg ctgaccggct gggccgccga gcagggcgcc gccgtcctcc 31380

gcagccacga cgtgacaggt gtgcgggaga ccgacgacgg cgtcgagctg gacgtggtca 31440gcagccacga cgtgacaggt gtgcgggaga ccgacgacgg cgtcgagctg gacgtggtca 31440

ctcccgaggg ccgcaagcag ctgcgtgccc gctacctggc cggttgcgac ggcggccgca 31500ctcccgaggg ccgcaagcag ctgcgtgccc gctacctggc cggttgcgac ggcggccgca 31500

gcaccgtccg caagctggtc ggcatcgact tccccggcgc cgacgccacg atcgagatgt 31560gcaccgtccg caagctggtc ggcatcgact tccccggcgc cgacgccacg atcgagatgt 31560

ggttcgccga cgtggccggc tgtgaactgc ggccccgttt ctccggcgag cgggtgccgg 31620ggttcgccga cgtggccggc tgtgaactgc ggccccgttt ctccggcgag cgggtgccgg 31620

gcgggatggt catggtgctg ccgctgggtc cgggggtcaa ccgtgtcgtc gtctacgagc 31680gcgggatggt catggtgctg ccgctgggtc cgggggtcaa ccgtgtcgtc gtctacgagc 31680

gcggcatgac ccggcagggc gacggcgccc cgagcttcgc cgaggtcgcc gctgcgtgga 31740gcggcatgac ccggcagggc gacggcgccc cgagcttcgc cgaggtcgcc gctgcgtgga 31740

accggctgac cggcgaggac atcagcggcg ggaagccgtt gtggacgagc tggacgaccg 31800accggctgac cggcgaggac atcagcggcg ggaagccgtt gtggacgagc tggacgaccg 31800

acgcgagccg gcaggcggcg gagtaccggc gcggacgcgt gttcctgctc ggcgacgccg 31860acgcgagccg gcaggcggcg gagtaccggc gcggacgcgt gttcctgctc ggcgacgccg 31860

cccacatcca cctgccggtg ggcgcgcagg ggatgagcgc gggggtcggc gacgccgtga 31920cccacatcca cctgccggtg ggcgcgcagg ggatgagcgc gggggtcggc gacgccgtga 31920

acctcggatg gaagctgggc gccgtgatca acggccacgc cccggacgac ctgctggaca 31980acctcggatg gaagctgggc gccgtgatca acggccacgc cccggacgac ctgctggaca 31980

cctaccacga cgagcggcac ccggtgggcg cccggatcct gaccaacacc ctcgcgcagc 32040cctaccacga cgagcggcac ccggtgggcg cccggatcct gaccaacacc ctcgcgcagc 32040

gcatcctgta cctgggtggc gacgagatgg acccgatgcg ggctgtcatg acggagctgc 32100gcatcctgta cctgggtggc gacgagatgg acccgatgcg ggctgtcatg acggagctgc 32100

tcgcgtacga ggaggtccag aagctgctgg tgggcatggt caccggcctg gacatccgct 32160tcgcgtacga ggaggtccag aagctgctgg tgggcatggt caccggcctg gacatccgct 32160

acgacgtcgg agcgggcagc cacccgctgc tcggccggcg gctgccggac gtcgagctca 32220acgacgtcgg agcgggcagc cacccgctgc tcggccggcg gctgccggac gtcgagctca 32220

ccggcgactt cggtccttcc ggcacgacgc gcgcgttcga actcctgcac tccggccggg 32280ccggcgactt cggtccttcc ggcacgacgc gcgcgttcga actcctgcac tccggccggg 32280

ccgtcgtcct ggacctggcc gacgacgcga agctgcgcgc cgcggccgag ccgtggaagg 32340ccgtcgtcct ggacctggcc gacgacgcga agctgcgcgc cgcggccgag ccgtggaagg 32340

accgggtcga cgtggtgacc ccggcgtcgc ggccgtccgg cgcgctggcc gacgtcgacg 32400accgggtcga cgtggtgacc ccggcgtcgc ggccgtccgg cgcgctggcc gacgtcgacg 32400

ccgtgctggt ccgcccggac ggctatgtcg cgtgggtccg tcccgacgcg gcggacggca 32460ccgtgctggt ccgcccggac ggctatgtcg cgtgggtccg tcccgacgcg gcggacggca 32460

ccgagctgcc cgacgccctg gcccggtggt tcggccgagc ctagccctct cgcggcgggc 32520ccgagctgcc cgacgccctg gcccggtggt tcggccgagc ctagccctct cgcggcgggc 32520

gccgccgcga tcacgtccat cagaggatgc gaggagaagc atgcccttca tcgaccccga 32580gccgccgcga tcacgtccat cagaggatgc gaggagaagc atgcccttca tcgaccccga 32580

gaacggctac ctgacggtca tcaacctgtt caagaccgac accccggagc gtgtgcaccg 32640gaacggctac ctgacggtca tcaacctgtt caagaccgac accccggagc gtgtgcaccg 32640

gctcgtcggc gagatgcggg cgatcgtcga cgtcgccgac taccccggct ggatctcgag 32700gctcgtcggc gagatgcggg cgatcgtcga cgtcgccgac taccccggct ggatctcgag 32700

caccgtgcac caggggcagg agcggcccgg caccgccaac ttcatccagt ggcgaggcaa 32760caccgtgcac caggggcagg agcggcccgg caccgccaac ttcatccagt ggcgaggcaa 32760

ccaggacctg gagtcgcggt acgccggcga cgagttcaag caccggacgg ttccggtgtt 32820ccaggacctg gagtcgcggt acgccggcga cgagttcaag caccggacgg ttccggtgtt 32820

ccacgagatc gccacgtaca tccggctgat gcagacggag gtcgagctga gtcagcgcca 32880ccacgagatc gccacgtaca tccggctgat gcagacggag gtcgagctga gtcagcgcca 32880

cccgtcactg ggtgacgtca cggagatctc accggaccgc gacgactaca cggtcatcga 32940cccgtcactg ggtgacgtca cggagatctc accggaccgc gacgactaca cggtcatcga 32940

gatcctcggg gtggcgcccg ccgaccagag tgagctgatc cgcgtccagg gcgccatgca 33000gatcctcggg gtggcgcccg ccgaccagag tgagctgatc cgcgtccagg gcgccatgca 33000

cgagtggctg gtggacgtgc cgggataccg ctcccagacc gtcctgcgcg gcatccggtc 33060cgagtggctg gtggacgtgc cgggataccg ctcccagacc gtcctgcgcg gcatccggtc 33060

gcgcggcgtc ggcggcaccg acgggggtct gacggccgtg ggtcaggaga aggacttcgt 33120gcgcggcgtc ggcggcaccg acgggggtct gacggccgtg ggtcaggaga aggacttcgt 33120

ggtggtctac tcgcagtggg acggcaagga gtcctacgac gcgttccgcg ccctgccctc 33180ggtggtctac tcgcagtggg acggcaagga gtcctacgac gcgttccgcg ccctgccctc 33180

ggcggatcag cccgccgcgc gccgcgcgtc gctggacaag cgggactcgc tggcgacctc 33240ggcggatcag cccgccgcgc gccgcgcgtc gctggacaag cgggactcgc tggcgacctc 33240

cgccgagtgg aacacctacc gcgtcgtgca ctcccggtcc gcggcgcaga ccacgcccgc 33300cgccgagtgg aacacctacc gcgtcgtgca ctcccggtcc gcggcgcaga ccacgcccgc 33300

ctgacccggt ccacacaccg ccggcggccc ctggcgttct cgccaggggc cgccgacggt 33360ctgacccggt ccacacaccg ccggcggccc ctggcgttct cgccaggggc cgccgacggt 33360

gcggctgtcg tcctatcgtg cggtgatcgt ctcccagtgc caccggctgt cgaacgcggt 33420gcggctgtcg tcctatcgtg cggtgatcgt ctcccagtgc caccggctgt cgaacgcggt 33420

gacgtcgagg ccggcgccgt cggcgaccag ccgggcggcg ctggtcgtcg cccgggcgat 33480gacgtcgagg ccggcgccgt cggcgaccag ccgggcggcg ctggtcgtcg cccgggcgat 33480

gaagcccgac gtgcgatgcg tggccagcag gtcctgctcg tcgaacggct tcccttcgct 33540gaagcccgac gtgcgatgcg tggccagcag gtcctgctcg tcgaacggct tcccttcgct 33540

gcgcagcagg cgcaggtcga cgaagccggt gaaggtacgt cccagcacga cgacgggctg 33600gcgcagcagg cgcaggtcga cgaagccggt gaaggtacgt cccagcacga cgacgggctg 33600

tggactgcgg atgctgaagc gcaccttggc ggcgacgtgg ccgtgatggt gctgatcgga 33660tggactgcgg atgctgaagc gcaccttggc ggcgacgtgg ccgtgatggt gctgatcgga 33660

gtggtaggcg atgtccggca tcgtgggcgg cacgaagtag tcccggtcca ccaccgccgg 33720gtggtaggcg atgtccggca tcgtgggcgg cacgaagtag tcccggtcca ccaccgccgg 33720

caccttcggc aggttgccga ccaggaagtg ccacgagttg tactgcatgc gcgcggcgat 33780caccttcggc aggttgccga ccaggaagtg ccacgagttg tactgcatgc gcgcggcgat 33780

ggaccacgcg gtgtcggcca ggtgggcggg ggagtccccg aagtaccggc gggcctgcgg 33840ggaccacgcg gtgtcggcca ggtgggcggg ggagtccccg aagtaccggc gggcctgcgg 33840

cgtcggaacc acgcagcaga agaagtgcgg gatgtcccac gccgtgatcg tggaccagct 33900cgtcggaacc acgcagcaga agaagtgcgg gatgtcccac gccgtgatcg tggaccagct 33900

ctcctcccgg agtgacgcga cgagccgggt cagcgagcgc atgccgcggc tcatcgcgaa 33960ctcctcccgg agtgacgcga cgagccgggt cagcgagcgc atgccgcggc tcatcgcgaa 33960

gtcggcgtcg aacaccgtgg tcgccgcgca caccgtctcg tacaccaggg actccagccc 34020gtcggcgtcg aacaccgtgg tcgccgcgca caccgtctcg tacaccaggg actccagccc 34020

cgtgccgaag tcgccgtgcg gctcgaccag ccagccgggt gacgcgtcga tgccggcacc 34080cgtgccgaag tcgccgtgcg gctcgaccag ccagccgggt gacgcgtcga tgccggcacc 34080

gagttccccg agtgtcgctc cggagacggg gagccggtca cgcaggcggg cgcagaggtc 34140gagttccccg agtgtcgctc cggagacggg gagccggtca cgcaggcggg cgcagaggtc 34140

cgcctcggct cgttccaccc tggcggtggc gggtccggcg agccgctcga ccttgtgcat 34200cgcctcggct cgttccaccc tggcggtggc gggtccggcg agccgctcga ccttgtgcat 34200

caacggtccg tggatctccc ggtagagctg aacccgcccg gcgatctcgc gtcggcgccg 34260caacggtccg tggatctccc ggtagagctg aacccgcccg gcgatctcgc gtcggcgccg 34260

ggcgaccgcc accgcgagcg cgtcgagggc cgccaccccg gccgaggcgg cggccggcac 34320ggcgaccgcc accgcgagcg cgtcgagggc cgccaccccg gccgaggcgg cggccggcac 34320

cgggtccccg ccggggaccg cgttgtacgc gacccgcgtg cgttccagga ggtgggccac 34380cgggtccccg ccggggaccg cgttgtacgc gacccgcgtg cgttccagga ggtgggccac 34380

gtggtcgagc gtcagctggg tgccgttggc ctcctcgatc cggccgtacc cggccgaccg 34440gtggtcgagc gtcagctggg tgccgttggc ctcctcgatc cggccgtacc cggccgaccg 34440

gaccagcagc gtgaggcaga ccaccatcag gacgtcgctg tccgaccaca gctcgagcgg 34500gaccagcagc gtgaggcaga ccaccatcag gacgtcgctg tccgaccaca gctcgagcgg 34500

tacggcgacc aggctgctca acgcgcagtc ggggtgcccg ggccacagcg tcttgccggt 34560tacggcgacc aggctgctca acgcgcagtc ggggtgcccg ggccacagcg tcttgccggt 34560

cagggtgttg tggtcgcgga agttggtgta ctgccggtcg ccgcagtaga acaccaccgg 34620cagggtgttg tggtcgcgga agttggtgta ctgccggtcg ccgcagtaga acaccaccgg 34620

cgcggtccgg cggaccgcct cggtgagctc ctccagcact ttctcccggc cgtccgggcc 34680cgcggtccgg cggaccgcct cggtgagctc ctccagcact ttctcccggc cgtccgggcc 34680

ggcgtcggcg agccagcggc ggcagtcggc gaccgagtcg cgcaggcgct cgtcggcttc 34740ggcgtcggcg agccagcggc ggcagtcggc gaccgagtcg cgcaggcgct cgtcggcttc 34740

ggcgagccgc gtccggtagt ccgccaggtc ctgcgcggac cagggtacgc gcagcacacc 34800ggcgagccgc gtccggtagt ccgccaggtc ctgcgcggac cagggtacgc gcagcacacc 34800

gccgacgagg tcgtcggtac tggtgagcgg acccctcgac ggcacccgga cgaccacccg 34860gccgacgagg tcgtcggtac tggtgagcgg acccctcgac ggcacccgga cgaccacccg 34860

gccgcccgtg gtcagcccga cctcgccgag gaaggccgcc tcgctgccac cggccggctg 34920gccgcccgtg gtcagcccga cctcgccgag gaaggccgcc tcgctgccac cggccggctg 34920

cggcgccacg ccgccgccgt cgtcgcgcca cgagcggccg aacaggatcc gcagtgcgca 34980cggcgccacg ccgccgccgt cgtcgcgcca cgagcggccg aacaggatcc gcagtgcgca 34980

ctcctccgcg gcgggcaggg cccggagctg gatgtccgcc gggacgtgcc cgtcgagggt 35040ctcctccgcg gcgggcaggg cccggagctg gatgtccgcc gggacgtgcc cgtcgagggt 35040

gagcagcgtg tccagggccg ccgccaggtc cggttcggtg gccgcgcgct gccagacccg 35100gagcagcgtg tccagggccg ccgccaggtc cggttcggtg gccgcgcgct gccagacccg 35100

cttcagcagg ccgggaaaac cggcgtcccc ggcgaccaga tagtcgggcc gggccgcggc 35160cttcagcagg ccgggaaaac cggcgtcccc ggcgaccaga tagtcgggcc gggccgcggc 35160

tccggcctgc cgcccgagcc ggctgggccg gcggctctgc cgtgccttgg ccagagcggc 35220tccggcctgc cgcccgagcc ggctgggccg gcggctctgc cgtgccttgg ccagagcggc 35220

tctgcgggag agcccgctat cgggctcgac cggcaaggtc atgctgcccc tccgcggtga 35280tctgcgggag agcccgctat cgggctcgac cggcaaggtc atgctgcccc tccgcggtga 35280

ggtcgatgtg gttccagagg ctgtcgtcgg tggccgtcca ctgctcgtcc acatagatgc 35340ggtcgatgtg gttccagagg ctgtcgtcgg tggccgtcca ctgctcgtcc acatagatgc 35340

agtcgaccgg gcagaccatc acgcacaccg ggcacccgga gcacagctcc gggatgatca 35400agtcgaccgg gcagaccatc acgcacaccg ggcacccgga gcacagctcc gggatgatca 35400

cgacgtccag cccccggtcg aagatcgccc cgaactcggg cgggcagctg cgcaggcacg 35460cgacgtccag cccccggtcg aagatcgccc cgaactcggg cgggcagctg cgcaggcacg 35460

tgtcgcacgt gatgcactcg gaagcctcga tgcggcgtgg cggcttcttc cagtcctcgt 35520tgtcgcacgt gatgcactcg gaagcctcga tgcggcgtgg cggcttcttc cagtcctcgt 35520

tcctggtccg ctgggcgatc cgggcggcgc gcgcggcgtc cgctccggca gcggtctcct 35580tcctggtccg ctgggcgatc cgggcggcgc gcgcggcgtc cgctccggca gcggtctcct 35580

ttgcccacgg catgtcctgt cctcacctcg tcttcctggt gtcacccggg cacgccggcg 35640ttgcccacgg catgtcctgt cctcacctcg tcttcctggt gtcacccggg cacgccggcg 35640

gccgcccggt acgaagcgca cggccgatca gcgggagcgc tcggagatct ccgcgaggat 35700gccgcccggt acgaagcgca cggccgatca gcgggagcgc tcggagatct ccgcgaggat 35700

gccgtcccag agccggaccc gggccgccag cgcggtgttg atggtgtcct cgcactgctc 35760gccgtcccag agccggaccc gggccgccag cgcggtgttg atggtgtcct cgcactgctc 35760

ccacttgtcg ttgtcgtccc cgcacaggtc ggcgagcatc tgcatcgcca tcggcgtgtg 35820ccacttgtcg ttgtcgtccc cgcacaggtc ggcgagcatc tgcatcgcca tcggcgtgtg 35820

ctcctcgctg tccacctgga tgtgccgccg caggtagtcg acgaaggtgg agagcacgcc 35880ctcctcgctg tccacctgga tgtgccgccg caggtagtcg acgaaggtgg agagcacgcc 35880

gacgccggcg tcgagcgcgg ccacctggtc gaacatgtcg ggaatcaggt cctcgcggcc 35940gacgccggcg tcgagcgcgg ccacctggtc gaacatgtcg ggaatcaggt cctcgcggcc 35940

gaacgcgaac gcggcggcct gacagtgcac gggcgcgccc tcgatgatcg tccaggtcgt 36000gaacgcgaac gcggcggcct gacagtgcac gggcgcgccc tcgatgatcg tccaggtcgt 36000

accgatgaac tcggccgaag gcccgggcac gccagccttc tgggcggcgg cccgcaccgg 36060accgatgaac tcggccgaag gcccgggcac gccagccttc tgggcggcgg cccgcaccgg 36060

ctcgccggcg cgcagcagac cgatgaactc gtcgatccgg gtggtgtcgg cgcccgcctg 36120ctcgccggcg cgcagcagac cgatgaactc gtcgatccgg gtggtgtcgg cgcccgcctg 36120

acgcatgccg tcgacgtaca gctcgaagtg gctgatgaag ccgtcgccga gttcgtcgct 36180acgcatgccg tcgacgtaca gctcgaagtg gctgatgaag ccgtcgccga gttcgtcgct 36180

ctcctcgacg agggtgatgt cgttgatgag ccggctgctg ccgggatgcc cggacggcac 36240ctcctcgacg agggtgatgt cgttgatgag ccggctgctg ccgggatgcc cggacggcac 36240

ccagggcacc tcgacgcagg tgagctgccg ctgcagcgac ttgagcaggg acatgaagtc 36300ccagggcacc tcgacgcagg tgagctgccg ctgcagcgac ttgagcaggg acatgaagtc 36300

ccagaccgcg aagacgtggt gctccatgaa ggtgaccacc gcgtcgacgg tgttcagctg 36360ccagaccgcg aagacgtggt gctccatgaa ggtgaccacc gcgtcgacgg tgttcagctg 36360

accgtagagc gggtgcccga tcaccttctg ccgggcgggc tcgacggcgt ccttgagctt 36420accgtagagc gggtgcccga tcaccttctg ccgggcgggc tcgacggcgt ccttgagctt 36420

gtcgatggcg ggggtggacc ctccccagtc gtaccgtgac atgaccgttc tcctttcgag 36480gtcgatggcg ggggtggacc ctccccagtc gtaccgtgac atgaccgttc tcctttcgag 36480

cgggtggggc cggcgtggtc agaagacacc gaagtactcg cggtgctccc attcggtgac 36540cgggtggggc cggcgtggtc agaagacacc gaagtactcg cggtgctccc attcggtgac 36540

gtggtcggcg ggcgggtgtt ccgtcgcgca ccaggcgcgg aagcgggacg cctcgctctc 36600gtggtcggcg ggcgggtgtt ccgtcgcgca ccaggcgcgg aagcgggacg cctcgctctc 36600

cttgagcttc gtcagacagg tcttcagcgg cgtgccgagc agctcctcgg ctcggctgct 36660cttgagcttc gtcagacagg tcttcagcgg cgtgccgagc agctcctcgg ctcggctgct 36660

cgcccggaac gcgtcgatcg cctcgtccag cgacgtcggc agcccgtcgg gctcaccgcc 36720cgcccggaac gcgtcgatcg cctcgtccag cgacgtcggc agcccgtcgg gctcaccgcc 36720

gcccggggcg gcgccggcgt ccaggccggc caggcccgcg tacagctgcg aggcgatgac 36780gcccggggcg gcgccggcgt ccaggccggc caggcccgcg tacagctgcg aggcgatgac 36780

caggtacgga ttcgcgcacg gttccccgac ccggttctcg acgtgcgtcg agctgccgcc 36840caggtacgga ttcgcgcacg gttccccgac ccggttctcg acgtgcgtcg agctgccgcc 36840

gctcagcacc cggaccatgg cgctgcggtc ctcctcgcgc cagtcggcgg cggtggggga 36900gctcagcacc cggaccatgg cgctgcggtc ctcctcgcgc cagtcggcgg cggtggggga 36900

gagcgtgaac cgcggcgcca gccggtggta gccgttcacg gtcgggacgg acagcaggca 36960gagcgtgaac cgcggcgcca gccggtggta gccgttcacg gtcgggacgg acagcaggca 36960

cagctcccgc gcgtgggcca gcaggccctg cacgtaggcc tcgcccgcgg gggagagcgg 37020cagctcccgc gcgtgggcca gcaggccctg cacgtaggcc tcgcccgcgg gggagagcgg 37020

acccgccgtg ccggcgtccg gcgcgaacag gttgcggctc gtcgccgtgc tcgtcaccga 37080acccgccgtg ccggcgtccg gcgcgaacag gttgcggctc gtcgccgtgc tcgtcaccga 37080

ctggtgcagg tgccagccgc tcgggtcgaa cccgtccagc cggggcagcg tcatgaacga 37140ctggtgcagg tgccagccgc tcgggtcgaa cccgtccagc cggggcagcg tcatgaacga 37140

cgcgtggtag ccgcggcgcc tgcactcctg cttgatctgg gtgcggaaca gcagcatcgc 37200cgcgtggtag ccgcggcgcc tgcactcctg cttgatctgg gtgcggaaca gcagcatcgc 37200

gtccgccgcg tccagcgcca gcatcgggtc gaacgtcgtc tcgagctgcc ccggaccgga 37260gtccgccgcg tccagcgcca gcatcgggtc gaacgtcgtc tcgagctgcc ccggaccgga 37260

ctcgtgctcg atcgtgcgca gcggcaggcc gagtgccatg agcatcgcgg ccaacgggtc 37320ctcgtgctcg atcgtgcgca gcggcaggcc gagtgccatg agcatcgcgg ccaacgggtc 37320

ggcgacgtcg gccaccgcgt cgtagtagct gtcgaggttg aactggtacc cggggttgac 37380ggcgacgtcg gccaccgcgt cgtagtagct gtcgaggttg aactggtacc cggggttgac 37380

ggcctccacc gacggggccg ggccctgccg gccgaatccg ttgccctcgt tgcccaccgg 37440ggcctccacc gacggggccg ggccctgccg gccgaatccg ttgccctcgt tgcccaccgg 37440

cccggcggtc cggcgcgtca ggtaccactc gacctcgagc ccgatcaccg gcacgaggtg 37500cccggcggtc cggcgcgtca ggtaccactc gacctcgagc ccgatcaccg gcacgaggtg 37500

ccggtcggcg tacgtggcgc acacccggcg caggacctcg cgggacgcca gtgggtgcgg 37560ccggtcggcg tacgtggcgc acacccggcg caggacctcg cgggacgcca gtgggtgcgg 37560

cgtgccgtcg cgcaggtatt cgttgccgag gacccaggcg gtccgttgct cgcggaccgg 37620cgtgccgtcg cgcaggtatt cgttgccgag gacccaggcg gtccgttgct cgcggaccgg 37620

cagcacctgg aacgtgcgcg gatccgggac cagcacgaag tcgcccgcgc cgagcagctc 37680cagcacctgg aacgtgcgcg gatccgggac cagcacgaag tcgcccgcgc cgagcagctc 37680

gccgacgccg acgccggagt cggcgaggaa gtccacggcg acggcatggc cggtgtcgaa 37740gccgacgccg acgccggagt cggcgaggaa gtccacggcg acggcatggc cggtgtcgaa 37740

gaggaacggt cccggactga agtccatgcc gttgcgcagc accgtgcgga agacgttcac 37800gaggaacggt cccggactga agtccatgcc gttgcgcagc accgtgcgga agacgttcac 37800

cggtacggtc ttggaccggg ccaggccgtg cgggtcgcag aagacgaccc ggacgagatc 37860cggtacggtc ttggaccggg ccaggccgtg cgggtcgcag aagacgaccc ggacgagatc 37860

cacgtcgtcg agctgcgcct cgaccttctc ggccgcggca gcctgctcgt cgtcccacag 37920cacgtcgtcg agctgcgcct cgaccttctc ggccgcggca gcctgctcgt cgtcccacag 37920

gccgaactcg gtcacgaacg aggggcgccc gacgccgctg ccgtcgccga gcagggacca 37980gccgaactcg gtcacgaacg aggggcgccc gacgccgctg ccgtcgccga gcagggacca 37980

tcgcctggac atcgtcatcg tcctccttga ctctcgcagc cggacggcag cgtcgtgtgg 38040tcgcctggac atcgtcatcg tcctccttga ctctcgcagc cggacggcag cgtcgtgtgg 38040

gccagttccg actcgacccg ggcggccagc cggagcacca ggtcgtcgtc gccgggccgt 38100gccagttccg actcgacccg ggcggccagc cggagcacca ggtcgtcgtc gccgggccgt 38100

cccacgagtt gcagcccgac cggcaacccg cgggacgtca ggccggccgg cagggacagc 38160cccacgagtt gcagcccgac cggcaacccg cgggacgtca ggccggccgg cagggacagc 38160

gccggctgac cggtgatgtt gaacgggtac gtcgccggcg accacgcgag ccacagcaga 38220gccggctgac cggtgatgtt gaacgggtac gtcgccggcg accacgcgag ccacagcaga 38220

tcggccgggt cggcggccca gggcggggcg acggcgtcgg cgtcgaacgg ctcgatcggg 38280tcggccgggt cggcggccca gggcggggcg acggcgtcgg cgtcgaacgg ctcgatcggg 38280

acggtcgcca tggccagcag gtcgtagcgt tccatcacgg cggcgagcgt cgcccgcagc 38340acggtcgcca tggccagcag gtcgtagcgt tccatcacgg cggcgagcgt cgcccgcagc 38340

gtcatccgca cctcctcggc ccgcatcacg gccgcgccgg tcagcgaccg cccgtaccgc 38400gtcatccgca cctcctcggc ccgcatcacg gccgcgccgg tcagcgaccg cccgtaccgc 38400

acgaccgcca gccggccctc gtcgcagagg tgctcgtcct ccggcgccgt accgtcggcc 38460acgaccgcca gccggccctc gtcgcagagg tgctcgtcct ccggcgccgt accgtcggcc 38460

tcggcggcgg cgacgacatc caccagggcc gcgtacgggt cggcgaacgg gacctcgatg 38520tcggcggcgg cgacgacatc caccagggcc gcgtacgggt cggcgaacgg gacctcgatg 38520

cgctcgacgc ggtggcccag gccggtcagc accgacggga cggcgtcggt cacgccgcgg 38580cgctcgacgc ggtggcccag gccggtcagc accgacggga cggcgtcggt cacgccgcgg 38580

acctcgtcgg acgttcccgg gaactccagc cagccgatcc gcagcgacgc gggcgccgcc 38640acctcgtcgg acgttcccgg gaactccagc cagccgatcc gcagcgacgc gggcgccgcc 38640

cgggacgtca agggggccgt ggacgagtcc gggtcccgct ggtccggccc ggtcagcacg 38700cgggacgtca agggggccgt ggacgagtcc gggtcccgct ggtccggccc ggtcagcacg 38700

gccatcagct cggcgacgtc ggcgacgctg cgggcgagcg gtcccacgtg cgccaggcgg 38760gccatcagct cggcgacgtc ggcgacgctg cgggcgagcg gtcccacgtg cgccaggcgg 38760

tcggcgcacg gcggtacgta ggggatccgg gcgaaggacg gcttgaaccc caccacaccg 38820tcggcgcacg gcggtacgta ggggatccgg gcgaaggacg gcttgaaccc caccacaccg 38820

cagaacgccg cggggatacg caccgagccg gccccgtccg tgccgatcga cgcggcgcac 38880cagaacgccg cggggatacg caccgagccg gccccgtccg tgccgatcga cgcggcgcac 38880

aggcccgccg ccaccgcggc ggcggatccg ccgctcgaac cgccggcggt ccggtccggt 38940aggcccgccg ccaccgcggc ggcggatccg ccgctcgaac cgccggcggt ccggtccggt 38940

gcccaggggt tccgcgtcgg cccggcgagc cggttcaccg tgctggcgct ccacccgtac 39000gcccaggggt tccgcgtcgg cccggcgagc cggttcaccg tgctggcgct ccacccgtac 39000

tccgacgtcg tcgtcttgcc cacgacgatc gcacccgccg cgcgcaggcg cgccacggcg 39060tccgacgtcg tcgtcttgcc cacgacgatc gcacccgccg cgcgcaggcg cgccacggcg 39060

ggcgcgtccg ccgccgcccg gcggttgggc aggagcgagc cccggcgggt gggaaggtct 39120ggcgcgtccg ccgccgcccg gcggttgggc aggagcgagc cccggcgggt gggaaggtct 39120

cgtgtctgga tcaggtcctt gacggagacc gggacgccga gcagcggccg ttcccggaag 39180cgtgtctgga tcaggtcctt gacggagacc gggacgccga gcagcggccg ttcccggaag 39180

gccgcctcgc cgacggtgcg gatcatccgg tcggccgcgt ccgcctcgcg cagtgcgccg 39240gccgcctcgc cgacggtgcg gatcatccgg tcggccgcgt ccgcctcgcg cagtgcgccg 39240

tcgcccgcca ccgcgacgaa cgccccgatt tccgggtcgg tgtcgcgaat ggcgtccagg 39300tcgcccgcca ccgcgacgaa cgccccgatt tccgggtcgg tgtcgcgaat ggcgtccagg 39300

acgttccgca cgtgcgccgc gaccgtcgaa cggcccgtga gaaacagttg tctcgtctcg 39360acgttccgca cgtgcgccgc gaccgtcgaa cggcccgtga gaaacagttg tctcgtctcg 39360

tggatgctgc cgcagaccgg ctgcctgata ctggaacccg tgaataccgg caccgcacca 39420tggatgctgc cgcagaccgg ctgcctgata ctggaacccg tgaataccgg caccgcacca 39420

ctacaggtca tcgacgcgta ttcgtgcaag tgttcgcgtg ccggtgggag ccggtcggag 39480ctacaggtca tcgacgcgta ttcgtgcaag tgttcgcgtg ccggtgggag ccggtcggag 39480

caatcgcgga gaagaaatgc ctcatccttc ggggtgggct ctgttcaact gctgaaaaat 39540caatcgcgga gaagaaatgc ctcatccttc ggggtgggct ctgttcaact gctgaaaaat 39540

ggtctaccag gcgaaagcac gcacggataa aacagaaaac acggctgtac agcacatcga 39600ggtctaccag gcgaaagcac gcacggataa aacagaaaac acggctgtac agcacatcga 39600

cggaatgact attctgcaca gtcatgattc cccgttattc gctgcccgag atggcggaca 39660cggaatgact attctgcaca gtcatgattc cccgttattc gctgcccgag atggcggaca 39660

tctggtcgga ccaggcgcgc tacgcgacat ggaccgaggt ggaggtcctg gccacgcagg 39720tctggtcgga ccaggcgcgc tacgcgacat ggaccgaggt ggaggtcctg gccacgcagg 39720

cgcaggcgat tctgggacgc gtaccggaga gcgccctcaa cgacatccgg agcgcccgtg 39780cgcaggcgat tctgggacgc gtaccggaga gcgccctcaa cgacatccgg agcgcccgtg 39780

tgccttcggt ggcccaggtg gtcgcgcacg agcgcgagcg cgaccacgag atcctcgcct 39840tgccttcggt ggcccaggtg gtcgcgcacg agcgcgagcg cgaccacgag atcctcgcct 39840

tcctcgcggc cctctgcgag ggcatcccgg aggactccgc ccggtgggtg cacctcggga 39900tcctcgcggc cctctgcgag ggcatcccgg aggactccgc ccggtgggtg cacctcggga 39900

tgaccagcta cgacctcgtc gacaccgccc agggctccac catggcccgg gcctgcgacc 39960tgaccagcta cgacctcgtc gacaccgccc agggctccac catggcccgg gcctgcgacc 39960

tgctgctcgg tgcggcggtg cggctccgcg cggtgctggc ggaccaggcg gtcgcgcact 40020tgctgctcgg tgcggcggtg cggctccgcg cggtgctggc ggaccaggcg gtcgcgcact 40020

ggcacacggt gtgcatcgga cggacccacg ggatgcacgc cgagcccacc acgctgggcc 40080ggcacacggt gtgcatcgga cggacccacg ggatgcacgc cgagcccacc acgctgggcc 40080

acaagttcgc cggcttcgcc ttcgcgatcg accggtccgt gcggcggctg cgggcggcac 40140acaagttcgc cggcttcgcc ttcgcgatcg accggtccgt gcggcggctg cgggcggcac 40140

gctcggaggt cgcggtcggg acgatctccg ggtccgtcgg gacctacgcg ctcatcgacc 40200gctcggaggt cgcggtcggg acgatctccg ggtccgtcgg gacctacgcg ctcatcgacc 40200

cgttcgtcga ggagtacgtc tgcgccgagc tgggcctcgg cgtggaaccc gcgccgaccc 40260cgttcgtcga ggagtacgtc tgcgccgagc tgggcctcgg cgtggaaccc gcgccgaccc 40260

aggtcgtggc gcgcgaccgg cacgcgcaac tcctgcacgt gatcgccctc ctcggcggat 40320aggtcgtggc gcgcgaccgg cacgcgcaac tcctgcacgt gatcgccctc ctcggcggat 40320

gcgtcgagca gatcgccacc gaggtccgcc tgctctcacg caccgagatc cgtgaggtgg 40380gcgtcgagca gatcgccacc gaggtccgcc tgctctcacg caccgagatc cgtgaggtgg 40380

aggagccccg gccggccgcc taccaggggt cgagcgccat gccgcacaag cgcaacccca 40440aggagccccg gccggccgcc taccaggggt cgagcgccat gccgcacaag cgcaacccca 40440

cgagcagcga gcgactggtg ggactggcgc gcctgctgcg cggatactcg gggacgatgc 40500cgagcagcga gcgactggtg ggactggcgc gcctgctgcg cggatactcg gggacgatgc 40500

tggagaacgt ggcgctgtgg cacgagcggg acctgtcgca ctcctcggtg gagcgggtcg 40560tggagaacgt ggcgctgtgg cacgagcggg acctgtcgca ctcctcggtg gagcgggtcg 40560

tcctgccgga cgcgatgatc gtcgcgcact accaggtgag cgcggcgacg gcgctggtgg 40620tcctgccgga cgcgatgatc gtcgcgcact accaggtgag cgcggcgacg gcgctggtgg 40620

cggggctcca ggtctttccc gaccgcatgc gggcggccgt cgacctcacc aacggcctgg 40680cggggctcca ggtctttccc gaccgcatgc gggcggccgt cgacctcacc aacggcctgg 40680

tctacagttc cgcagtcctg gccgacctgc tggagcgcgg caccgagcgg gaacgtgcct 40740tctacagttc cgcagtcctg gccgacctgc tggagcgcgg caccgagcgg gaacgtgcct 40740

accgggcggt ccaggccgcc gccgacggag ccggctccgg cgacgccgac ttcgggaccc 40800accgggcggt ccaggccgcc gccgacggag ccggctccgg cgacgccgac ttcgggaccc 40800

tgctccgggc ccagggggtc gacgtcggcc cgcttcggcc ggaccgtttc ctcgtccacc 40860tgctccgggc ccagggggtc gacgtcggcc cgcttcggcc ggaccgtttc ctcgtccacc 40860

acgatgtcgt cctcaagcga ttggaaagcc tccgtgaact ggaagattga acgcgagacg 40920acgatgtcgt cctcaagcga ttggaaagcc tccgtgaact ggaagattga acgcgagacg 40920

ggcgccgacc tggatatgga agcggtcctg ggcgtctacc ggtcctccgg gctgggcgag 40980ggcgccgacc tggatatgga agcggtcctg ggcgtctacc ggtcctccgg gctgggcgag 40980

cgccggccca ttgctgacgt cgcgcgaatg gccaccatgg tgagatcggc gaacctgatc 41040cgccggccca ttgctgacgt cgcgcgaatg gccaccatgg tgagatcggc gaacctgatc 41040

gtgacctgcc gaatcgaggg tgagctggtg ggcatcgcga ggagtgtttc cgacttctcg 41100gtgacctgcc gaatcgaggg tgagctggtg ggcatcgcga ggagtgtttc cgacttctcg 41100

tacgtgacct acctctcgga catcgccgtc gcgcgttccc atcagcgctc cggtatcggc 41160tacgtgacct acctctcgga catcgccgtc gcgcgttccc atcagcgctc cggtatcggc 41160

aaggccctca tcgaggcgac ccggaacgag gcgccgatgg ccaagatcgt gctcctgtcg 41220aaggccctca tcgaggcgac ccggaacgag gcgccgatgg ccaagatcgt gctcctgtcg 41220

gcgcccgccg cgagcgacta ctacccgcac atcggtttca cccggcacga ttccgcctgg 41280gcgcccgccg cgagcgacta ctacccgcac atcggtttca cccggcacga ttccgcctgg 41280

gtgctcaatc cgtagcaagg caggcccgcg gtgccggacc cgcacggcga aacggccggt 41340gtgctcaatc cgtagcaagg caggcccgcg gtgccggacc cgcacggcga aacggccggt 41340

gcgggtaccg gcgtgccgcg ggccgggcgc gtgatccgcg cccggcccga ccggtggcgg 41400gcgggtaccg gcgtgccgcg ggccgggcgc gtgatccgcg cccggcccga ccggtggcgg 41400

tcccgtcagc gggtctcggc gggaagctgg gcgtcgcggc tgatcagcat cgcgtagtgg 41460tcccgtcagc gggtctcggc gggaagctgg gcgtcgcggc tgatcagcat cgcgtagtgg 41460

cccccctgcg ccagcagttc gtcgtgggta ccgcgctcga cgatcgtgcc ggcgtccaac 41520cccccctgcg ccagcagttc gtcgtgggta ccgcgctcga cgatcgtgcc ggcgtccaac 41520

acgatgatct cgtcggcgtc ccggatcgtc gacagccggt gggcgatcgt gactgtggtg 41580acgatgatct cgtcggcgtc ccggatcgtc gacagccggt gggcgatcgt gactgtggtg 41580

cggccggcgc tcgccgcctc gatggcctgg tgcacggcct gctcggtctg gttgtccaag 41640cggccggcgc tcgccgcctc gatggcctgg tgcacggcct gctcggtctg gttgtccaag 41640

gcgctcgtgg cttcgtccag cacgagaatc ggcgggtcgc gcaggaccgc gcgggcgatc 41700gcgctcgtgg cttcgtccag cacgagaatc ggcgggtcgc gcaggaccgc gcgggcgatc 41700

gccaggcgct gccgctcccc gccggagaac cggaatccgc gttcgcccac caccgtgtcg 41760gccaggcgct gccgctcccc gccggagaac cggaatccgc gttcgcccac caccgtgtcg 41760

tagccgtcgg gaagagacat caggtggtcg tggatgtgcg cgatcttcgc ggccgcgacg 41820tagccgtcgg gaagagacat caggtggtcg tggatgtgcg cgatcttcgc ggccgcgacg 41820

agttcgtcgt cggtcgcccc gggccgggcg aaccgcaggt tctcggcgac cgaggtgtgg 41880agttcgtcgt cggtcgcccc gggccgggcg aaccgcaggt tctcggcgac cgaggtgtgg 41880

aacaggtagg tctcctggaa gaccatcccg accgccccgg cgagcgagtc ggcggacatg 41940aacaggtagg tctcctggaa gaccatcccg accgccccgg cgagcgagtc ggcggacatg 41940

tcccgcacgt cgacgccgtc gatggtcacc cgcccggacg tgacgtcgta cagccgcgga 42000tcccgcacgt cgacgccgtc gatggtcacc cgcccggacg tgacgtcgta cagccgcgga 42000

atcaggtaac tgagggtggt cttgccggat ccggtctcgc cgacgatggc gagcttgcgc 42060atcaggtaac tgagggtggt cttgccggat ccggtctcgc cgacgatggc gagcttgcgc 42060

ccggcgggaa cggtgatgtc tatggcgttg agcgtgggcc gggcggcgcc cgggtacgcg 42120ccggcgggaa cggtgatgtc tatggcgttg agcgtgggcc gggcggcgcc cgggtacgcg 42120

aagcagaccc cctgcatccg cagttcgccg cgcggcttca ccgtgttggc gtgccggggc 42180aagcagaccc cctgcatccg cagttcgccg cgcggcttca ccgtgttggc gtgccggggc 42180

tcgctgatct ccaccggcag gtcgaggtac tcgaagatcc ggttgaacag ttcgagcgag 42240tcgctgatct ccaccggcag gtcgaggtac tcgaagatcc ggttgaacag ttcgagcgag 42240

gtctgcagct ccacggcgac atccagcagg gacaccgccg gtcgcagcag gttctgttgc 42300gtctgcagct ccacggcgac atccagcagg gacaccgccg gtcgcagcag gttctgttgc 42300

agcgtggtga aggcgaccaa cgtaccgatg gagatcgcca tgctgtcgcc ccggccgctc 42360agcgtggtga aggcgaccaa cgtaccgatg gagatcgcca tgctgtcgcc ccggccgctc 42360

tgccccgcca cccagtagat caccgccggc atcgacgcca tgacgaccca cgtcgtcgcc 42420tgccccgcca cccagtagat caccgccggc atcgacgcca tgacgaccca cgtcgtcgcc 42420

tggctccacc ggcccgccgt ctgcgagcgg atctccagat ccgacagggt ccgggactcg 42480tggctccacc ggcccgccgt ctgcgagcgg atctccagat ccgacagggt ccgggactcg 42480

gcggtgaagg cgtcggtcag cgagcgggag cgtcccatcg tccgggccag cagcacgcca 42540gcggtgaagg cgtcggtcag cgagcgggag cgtcccatcg tccgggccag cagcacgcca 42540

ctgacggaca acgactcctg cacgatcgcg gcgatcgtcg cgtactggcg ttgccgctcc 42600ctgacggaca acgactcctg cacgatcgcg gcgatcgtcg cgtactggcg ttgccgctcc 42600

cgggtgatgc ggcgccgctg gttgcccacc cggcggccca cccacacgaa gaacggcacc 42660cgggtgatgc ggcgccgctg gttgcccacc cggcggccca cccacacgaa gaacggcacc 42660

atcaccagcg acacgatggt gagacgccag tcgaggatgg ccatcgcgac gacggtggcc 42720atcaccagcg acacgatggt gagacgccag tcgaggatgg ccatcgcgac gacggtggcc 42720

accacgactg tcacgtcgga caccagggtc gtcgcggacg tggtcacggt ggcctgcatg 42780accacgactg tcacgtcgga caccagggtc gtcgcggacg tggtcacggt ggcctgcatg 42780

tcgccgatgt cgttggtgat ccgcgactgc acctcgccgg cgcgggtccg cgtgtagaag 42840tcgccgatgt cgttggtgat ccgcgactgc acctcgccgg cgcgggtccg cgtgtagaag 42840

gccagcggca tgcgctgcag gtgcgcgtag acagccgatc gcaggtcgtg catgacctgc 42900gccagcggca tgcgctgcag gtgcgcgtag acagccgatc gcaggtcgtg catgacctgc 42900

tcgccgacct tcgtcgacac gtacgtctgc cagacgacga aggagctgtt cgcgaccgag 42960tcgccgacct tcgtcgacac gtacgtctgc cagacgacga aggagctgtt cgcgaccgag 42960

gcgaggatca tgccgagggc gaggacggtc agcaggcccg tccgtccctg gggcagggcc 43020gcgaggatca tgccgagggc gaggacggtc agcaggcccg tccgtccctg gggcagggcc 43020

acgtccagga tctcgcggat caggaacggg ttgacgagga agaccgctga ggtggcgcag 43080acgtccagga tctcgcggat caggaacggg ttgacgagga agaccgctga ggtggcgcag 43080

acgagagcgg cgacgagcag caggctccat ccgtacgggc ggaagagccg gaggatgcgc 43140acgagagcgg cgacgagcag caggctccat ccgtacgggc ggaagagccg gaggatgcgc 43140

cgcaacggca catgggggtc agcggtggtc accaggcctc ctcagccgac gtgcgtcagg 43200cgcaacggca catgggggtc agcggtggtc accaggcctc ctcagccgac gtgcgtcagg 43200

cgcggcgcgc caccggagga gtcgtcgcgc cctcgggcgc gacggggcgg catgctccgg 43260cgcggcgcgc caccggagga gtcgtcgcgc cctcgggcgc gacggggcgg catgctccgg 43260

acgagaacca tatgtcgtgc aagcccgcgg atcgcggcgt cggacggcag tgcgcacagc 43320acgagaacca tatgtcgtgc aagcccgcgg atcgcggcgt cggacggcag tgcgcacagc 43320

cggggatccg ctaccgcagg cgctgctcca gggccgccca ctcggcgcgt tcggccgctg 43380cggggatccg ctaccgcagg cgctgctcca gggccgccca ctcggcgcgt tcggccgctg 43380

tcagcggctc gtccatgtgc atcgtcacga atcccgtcgg cacgccggcc cacatggctc 43440tcagcggctc gtccatgtgc atcgtcacga atcccgtcgg cacgccggcc cacatggctc 43440

ccgaccagat caggcccatc ttgaggtgat gcagcatgga acgcagcccg gccggcagcc 43500ccgaccagat caggcccatc ttgaggtgat gcagcatgga acgcagcccg gccggcagcc 43500

gtggggtctt catgatgggc tctcctcatg cgagtcgatg gacgagggcg gcgccgccgc 43560gtggggtctt catgatgggc tctcctcatg cgagtcgatg gacgagggcg gcgccgccgc 43560

gcgaccgctc cccatcaggg ctgccgctcg gcggtccggg tgagggcgac gtagacgtat 43620gcgaccgctc cccatcaggg ctgccgctcg gcggtccggg tgagggcgac gtagacgtat 43620

ccgtcctcca cccgcacggg gtaggtgccg atcggccggg tcgcgggtgg gttggtgggc 43680ccgtcctcca cccgcacggg gtaggtgccg atcggccggg tcgcgggtgg gttggtgggc 43680

tcaccggtcc gcaggtcgaa gcaggatccg tgcagccagc actcgatcgt gccgtcgtac 43740tcaccggtcc gcaggtcgaa gcaggatccg tgcagccagc actcgatcgt gccgtcgtac 43740

acctcgccct cggagagcga gacctcggcg tgcgagcaca ggtcgtggat cgcgtacacc 43800acctcgccct cggagagcga gacctcggcg tgcgagcaca ggtcgtggat cgcgtacacc 43800

tgcccgccgc tgcgggccag ggccaccgtg gtgccctcga tctccagcgc gatcgcgccc 43860tgcccgccgc tgcgggccag ggccaccgtg gtgccctcga tctccagcgc gatcgcgccc 43860

tcatccggga cgtctgccac cggacacgcc ttgacgaaat ccatcgagtc agccctctcc 43920tcatccggga cgtctgccac cggacacgcc ttgacgaaat ccatcgagtc agccctctcc 43920

cgatcgcacc cggacggaaa cgaccatcgc caccttgcgc ccgggccctc aggcaggaac 43980cgatcgcacc cggacggaaa cgaccatcgc caccttgcgc ccgggccctc aggcaggaac 43980

ggagggcggg tcgaaccgcg ctggaggcgc cgccgccggg ggcggcgcgg gctcgttcgg 44040ggagggcggg tcgaaccgcg ctggaggcgc cgccgccggg ggcggcgcgg gctcgttcgg 44040

gtggtccctc gggcggcgga ggagccctcg tggaaccgac ggaaagggac cagaagcgtc 44100gtggtccctc gggcggcgga ggagccctcg tggaaccgac ggaaagggac cagaagcgtc 44100

aggaatcaag aagcactggt cgcgcggcgg ccgctagcgt ttgcggtatg aacatcacaa 44160aggaatcaag aagcactggt cgcgcggcgg ccgctagcgt ttgcggtatg aacatcacaa 44160

ttcactcaag cttcctgccg cacgaggatg cggacgcgtc ggtcaccttc tatcgcgacg 44220ttcactcaag cttcctgccg cacgaggatg cggacgcgtc ggtcaccttc tatcgcgacg 44220

tgctggggtt cgagatccgc aacgacgtgg gctacgacgg catgcgctgg gtcaccgtgg 44280tgctggggtt cgagatccgc aacgacgtgg gctacgacgg catgcgctgg gtcaccgtgg 44280

ggccggccga ccagccgcag acgtcgatcg tgctgcagcc gccggccgcc gaccccggca 44340ggccggccga ccagccgcag acgtcgatcg tgctgcagcc gccggccgcc gaccccggca 44340

tcaccgacga ggagcggcgc accatccagg agatgatggc caagggcacg tacgccggtg 44400tcaccgacga ggagcggcgc accatccagg agatgatggc caagggcacg tacgccggtg 44400

tgctcctgtc cgctcccgac ctcgacgggg tcttcgagcg cgtccaggcc agcggggccg 44460tgctcctgtc cgctcccgac ctcgacgggg tcttcgagcg cgtccaggcc agcggggccg 44460

acgtggtgca ggagccgatc gaacaggact acggcgtgcg ggactgcgcg ttccgcgacc 44520acgtggtgca ggagccgatc gaacaggact acggcgtgcg ggactgcgcg ttccgcgacc 44520

cggccggcaa tttgatcagg attcagcagg cacgctgagc cccaccctgg ttctcttcgt 44580cggccggcaa tttgatcagg attcagcagg cacgctgagc cccaccctgg ttctcttcgt 44580

caatcagggc cgggaaccgg tcccggcccg catcgtcatc tcagccagag gaggtcgtca 44640caatcagggc cgggaaccgg tcccggcccg catcgtcatc tcagccagag gaggtcgtca 44640

catgctgttg cccgacgtga tcacgatcgg ggcgacggac ccgcaagccg cccatgcctt 44700catgctgttg cccgacgtga tcacgatcgg ggcgacggac ccgcaagccg cccatgcctt 44700

ctacaagtcc gtgttcctgc ccaccggggt cgagcaccgg gacgagcggg tcgagctcga 44760ctacaagtcc gtgttcctgc ccaccggggt cgagcaccgg gacgagcggg tcgagctcga 44760

cctgcacggt gtcgcacagc tggccgtgtg cccatccgag gcgctggccg cggaggcgaa 44820cctgcacggt gtcgcacagc tggccgtgtg cccatccgag gcgctggccg cggaggcgaa 44820

cgtggagccc aaggcgcccg gttatcgcgg ctacgtcctc acctacatcg tcgaccagcc 44880cgtggagccc aaggcgcccg gttatcgcgg ctacgtcctc acctacatcg tcgaccagcc 44880

cagcgaggtg aggagcattc tcgacgccgc cgcccagcac ggcgcgagcg tcctcaagcc 44940cagcgaggtg aggagcattc tcgacgccgc cgcccagcac ggcgcgagcg tcctcaagcc 44940

cgcgaagaag gccatgttcg ccggcttctc cggtgtgttc caggcgccgg acggcgcggt 45000cgcgaagaag gccatgttcg ccggcttctc cggtgtgttc caggcgccgg acggcgcggt 45000

gtggaagctc gcgtcgtccg agaagaagga caccggcccg gcgcaggaga cgcccgtgcc 45060gtggaagctc gcgtcgtccg agaagaagga caccggcccg gcgcaggaga cgcccgtgcc 45060

gagcgaggtc ggcgtgctgc tcggggtggc cgacacgaag gcgtccacgg ccttctacca 45120gagcgaggtc ggcgtgctgc tcggggtggc cgacacgaag gcgtccacgg ccttctacca 45120

ggccctgggc atgaccgtcg accgcgacta cggcagcaag tacatcgact tcaagccgac 45180ggccctgggc atgaccgtcg accgcgacta cggcagcaag tacatcgact tcaagccgac 45180

ctcgggcggc tgccggctgg gcctgatgga cggcggcatg ctggcgaagg acgccggggt 45240ctcgggcggc tgccggctgg gcctgatgga cggcggcatg ctggcgaagg acgccggggt 45240

caaggagagc ggcggcgccg gcttccgcgc ggcggccttc gagcgggccg cggcgtcgcg 45300caaggagagc ggcggcgccg gcttccgcgc ggcggccttc gagcgggccg cggcgtcgcg 45300

cgatgaggtc gacagccttc tggccgcggt gactccggcc ggcggccacg tcgccgcagc 45360cgatgaggtc gacagccttc tggccgcggt gactccggcc ggcggccacg tcgccgcagc 45360

cggcgcggag acctcgccgg gggtctactc gggatacttc gtcgacccgg acggcttcct 45420cggcgcggag acctcgccgg gggtctactc gggatacttc gtcgacccgg acggcttcct 45420

gtggagggcg accagcgcat agcagagcgg gcctctggcc gggatgtctc ccggccagag 45480gtggagggcg accagcgcat agcagagcgg gcctctggcc gggatgtctc ccggccagag 45480

gcccggtgcc gtgctcagat cagccgtcag ccggatccgg caacggattg gcgttgcagt 45540gcccggtgcc gtgctcagat cagccgtcag ccggatccgg caacggattg gcgttgcagt 45540

gggcgtcgac gccggcgacc gaattcggcc gcgtcccgtc cctcaggtag gcgatgacgt 45600gggcgtcgac gccggcgacc gaattcggcc gcgtcccgtc cctcaggtag gcgatgacgt 45600

ggttgttcag gcacgcgttg gcgttggcgg tcagggacgc gccgtggaag acgccgccac 45660ggttgttcag gcacgcgttg gcgttggcgg tcagggacgc gccgtggaag acgccgccac 45660

gctccaggac cagccgggag ttcgggaaca accggtgtac ctcgacggcg ccctgcaccg 45720gctccaggac cagccgggag ttcgggaaca accggtgtac ctcgacggcg ccctgcaccg 45720

gcgtgagggc gtcgtgttcc gcctggatca gcagcatgtt cacgttccgg tcgccgacct 45780gcgtgagggc gtcgtgttcc gcctggatca gcagcatgtt cacgttccgg tcgccgacct 45780

tcgtcggctt gcccgccggc accggccaga aggcgcacgg cgcgttgtac cacccgttgt 45840tcgtcggctt gcccgccggc accggccaga aggcgcacgg cgcgttgtac cacccgttgt 45840

tccaggtcat gaacttgttg cccctgcggt gctgctggga cagatcggtc cgccactggt 45900tccaggtcat gaacttgttg cccctgcggt gctgctggga cagatcggtc cgccactggt 45900

tccagttccg gggccaggga ccgtcgacgc actgcacggc gcggtacatc gcgtgggtgt 45960tccagttccg gggccaggga ccgtcgacgc actgcacggc gcggtacatc gcgtgggtgt 45960

tctgggcggg gaagtcgggc tcgaggaagt tgtcccgaag cccggacggg tccccgcgca 46020tctgggcggg gaagtcgggc tcgaggaagt tgtcccgaag cccggacggg tccccgcgca 46020

gtacccagtc ggccagcacc ccggcgtggt aggtccagat gtagctgcga tacacgttca 46080gtacccagtc ggccagcacc ccggcgtggt aggtccagat gtagctgcga tacacgttca 46080

ccgagaagat gtcgctgtac tcggccggcc cgatcttgcc gtcgatcggc gccttgcgga 46140ccgagaagat gtcgctgtac tcggccggcc cgatcttgcc gtcgatcggc gccttgcgga 46140

gcatcgccat gcccttgtag tagttcgcct cgacctgtgc cggcgtccgt ccgaggtggt 46200gcatcgccat gcccttgtag tagttcgcct cgacctgtgc cggcgtccgt ccgaggtggt 46200

agtacgagtc gtgctcggcg atccaggcga agtagatctg ggcccgcttc tcgaaggctt 46260agtacgagtc gtgctcggcg atccaggcga agtagatctg ggcccgcttc tcgaaggctt 46260

cgttcgtctc cagcagggat ttgtacccga cgctgctcgg ccgcaccacg ctgtccagga 46320cgttcgtctc cagcagggat ttgtacccga cgctgctcgg ccgcaccacg ctgtccagga 46320

ccatgcgccg tacccggttc gggaacaggg aggcgtagac ggagccgacg taggtgccgt 46380ccatgcgccg tacccggttc gggaacaggg aggcgtagac ggagccgacg taggtgccgt 46380

gggagtagcc caggtaattg attttctcct ggccgagggc gatccgcatg aggtccatgt 46440gggagtagcc caggtaattg attttctcct ggccgagggc gatccgcatg aggtccatgt 46440

cgcgggcggt gtccgccgtc cggaagtact tgagcgtgtc accgtacttc tgcccgcagc 46500cgcgggcggt gtccgccgtc cggaagtact tgagcgtgtc accgtacttc tgcccgcagc 46500

tctcggcgaa tgtgcgcgcc cggttcaccc acaccgcttc ctcggcgttg ccggccggaa 46560tctcggcgaa tgtgcgcgcc cggttcaccc acaccgcttc ctcggcgttg ccggccggaa 46560

cgtagtcggg ccgggccccg cccgggtaga ggtacgtggg atcgcagctg atcgacggct 46620cgtagtcggg ccgggccccg cccgggtaga ggtacgtggg atcgcagctg atcgacggct 46620

cgctcgcgcc gacgccgcgc gggtcgaagc cgatccagtc gtacgtggag cccacggagg 46680cgctcgcgcc gacgccgcgc gggtcgaagc cgatccagtc gtacgtggag cccacggagg 46680

tcagcaggcc ggtggagccg tccgcgaagc gggtcggcag atcccgtccg atggtgcccg 46740tcagcaggcc ggtggagccg tccgcgaagc gggtcggcag atcccgtccg atggtgcccg 46740

gccactggcc ccggttcaac aggacgacgc cctggtactg cgattcgggc gaggtgtgct 46800gccactggcc ccggttcaac aggacgacgc cctggtactg cgattcgggc gaggtgtgct 46800

tggcccgggt cagggccaga gtgatcttct ggccgttcgg ccgggagtgg tcgagtggaa 46860tggcccgggt cagggccaga gtgatcttct ggccgttcgg ccgggagtgg tcgagtggaa 46860

cctcgaggga accgcactcg agccccacga gatagtcgtc catcagaggg tcgtcgtccg 46920cctcgaggga accgcactcg agccccacga gatagtcgtc catcagaggg tcgtcgtccg 46920

gacaaggtga ccaggcgatt ccggcaatcg gccggtcggc cggccgggcc gccgccgcgg 46980gacaaggtga ccaggcgatt ccggcaatcg gccggtcggc cggccgggcc gccgccgcgg 46980

aggggaccac ggtgacctgt gccgtcagac ccagcgcaac cgccgccgcg ccggctaaca 47040aggggaccac ggtgacctgt gccgtcagac ccagcgcaac cgccgccgcg ccggctaaca 47040

gttttctcat ccattccccc attgctcgaa agcgaatgca ttcacagatg cgtggccgct 47100gttttctcat ccattccccc attgctcgaa agcgaatgca ttcacagatg cgtggccgct 47100

gattcgaccg ggctcgacgc gctttgtcaa accagcttca caggtcgaat caatgagttc 47160gattcgaccg ggctcgacgc gctttgtcaa accagcttca caggtcgaat caatgagttc 47160

gtgcggcggc gattcggcgg gggaatgcgg cacgtaacgc tgcaacagcg cctattcggg 47220gtgcggcggc gattcggcgg gggaatgcgg cacgtaacgc tgcaacagcg cctattcggg 47220

tcttacctcg ccagtccggc cattcgaatc gtcccaggct gacgccggtg cggagggaag 47280tcttacctcg ccagtccggc cattcgaatc gtcccaggct gacgccggtg cggagggaag 47280

cgctgtcgga ggtcgtggtg cggccatcga tcggtcggcc gacggcgggg tgcccggccg 47340cgctgtcgga ggtcgtggtg cggccatcga tcggtcggcc gacggcgggg tgcccggccg 47340

gccgacatgg ctggcggaaa atgcacagtg atggcgacga ggagcggttc ggcctgccga 47400gccgacatgg ctggcggaaa atgcacagtg atggcgacga ggagcggttc ggcctgccga 47400

cccggtcggt cgatcatgcg gatcgggtcg attcttcagt ccttacggca cgggcgtccg 47460cccggtcggt cgatcatgcg gatcgggtcg attcttcagt ccttacggca cgggcgtccg 47460

gaggcggtga cggccgaggg gaattcaaat aactcgatgc cggcggattc ggtccacacg 47520gaggcggtga cggccgaggg gaattcaaat aactcgatgc cggcggattc ggtccacacg 47520

aagggacccg gccgagggcg actgccccgg ccgggtccct gtcggaccgg tcacccggtg 47580aagggacccg gccgagggcg actgccccgg ccgggtccct gtcggaccgg tcacccggtg 47580

accagcgact ccatccgtcg gaccacatcg gccggcgtcg ggagcgcggc gatctcggcg 47640accagcgact ccatccgtcg gaccacatcg gccggcgtcg ggagcgcggc gatctcggcg 47640

gccagggctc gcgcccgccg ggagtaccgc gggtcggcga ggatctcccg gctgtccgcg 47700gccagggctc gcgcccgccg ggagtaccgc gggtcggcga ggatctcccg gctgtccgcg 47700

gccatggcgt cagccaggtc atggctcccc tccggcagct ccgaccggat cgtccgcgcc 47760gccatggcgt cagccaggtc atggctcccc tccggcagct ccgaccggat cgtccgcgcc 47760

gcgcctgcgg ccgacaggac gccggcgatc gccctggtgt gcgtgttcgg gggagtgatc 47820gcgcctgcgg ccgacaggac gccggcgatc gccctggtgt gcgtgttcgg gggagtgatc 47820

agctggggga cccccgccgt catgatggtc atcgcggtgg tcgccccgcc gtggtgcacc 47880agctggggga cccccgccgt catgatggtc atcgcggtgg tcgccccgcc gtggtgcacc 47880

gtcaggtcgc aggtggggac gacgacgtcc agcgggatcc agccgacgcg gacgtcgtcc 47940gtcaggtcgc aggtggggac gacgacgtcc agcgggatcc agccgacgcg gacgtcgtcc 47940

gtgtccgccc cgaactccgc ggcgacctcc gcgtgcgccg cgatcaccac ctcggctccc 48000gtgtccgccc cgaactccgc ggcgacctcc gcgtgcgccg cgatcaccac ctcggctccc 48000

gcggaggtga gctgccgcac caggtggtgc agcgcgctgc cggcgagcat gcggtggtgc 48060gcggaggtga gctgccgcac caggtggtgc agcgcgctgc cggcgagcat gcggtggtgc 48060

gtcccggagg tgatcagcac ccggggccgg tccgtcggcc gggtgtacat ccatcgttcg 48120gtcccggagg tgatcagcac ccggggccgg tccgtcggcc gggtgtacat ccatcgttcg 48120

aggcggcgct gaggattcgt cgcgacgaaa cccatgggct gcgcccgcgc cgacggcgac 48180aggcggcgct gaggattcgt cgcgacgaaa cccatgggct gcgcccgcgc cgacggcgac 48180

ggcccgaggg acggcggagt gacgtcgatc agcatcgcgg gatccgggag cccgcccagg 48240ggcccgaggg acggcggagt gacgtcgatc agcatcgcgg gatccgggag cccgcccagg 48240

cccaggtcct tcagctgcgg cgccagctcc tcctcggccg ccggatcgat ctccgtcatc 48300cccaggtcct tcagctgcgg cgccagctcc tcctcggccg ccggatcgat ctccgtcatc 48300

gggatctgga ggtactcggc gtgccggacg aaggggacgc ccagccgggt ggcgagcagg 48360gggatctgga ggtactcggc gtgccggacg aaggggacgc ccagccgggt ggcgagcagg 48360

cccgccgcgt agctcatcga actcccgagg accacgtcgg gagcccagtc ctccgcgagg 48420cccgccgcgt agctcatcga actcccgagg accacgtcgg gagcccagtc ctccgcgagg 48420

tcccgcaggg tgggcgtcgc ggcggcggcc atcctggcca ggccgcggcc ggcgaccacc 48480tcccgcaggg tgggcgtcgc ggcggcggcc atcctggcca ggccgcggcc ggcgaccacc 48480

atctcgtccg tcgagccgac gccggtcagg aagtgccgga tcggctccgg catgatcgac 48540atctcgtccg tcgagccgac gccggtcagg aagtgccgga tcggctccgg catgatcgac 48540

actccgggta cgccgatgcc ctccaccgca tcgatcagcg gttcgttcgt ggccaccagg 48600actccgggta cgccgatgcc ctccaccgca tcgatcagcg gttcgttcgt ggccaccagg 48600

agctcgtgcc ccgcgttgcg caccgccttc gccagagcgg cggtcgcgta gacggtgacg 48660agctcgtgcc ccgcgttgcg caccgccttc gccagagcgg cggtcgcgta gacggtgacg 48660

tggctgccgg tggtgaggaa caggaatctc atgctcggtc gccggccttc cgcgcggggc 48720tggctgccgg tggtgaggaa caggaatctc atgctcggtc gccggccttc cgcgcggggc 48720

tgccggcctc cgggcggcac tcgaccacga actggccgga cggcgtgcgc ccgtacgcgg 48780tgccggcctc cgggcggcac tcgaccacga actggccgga cggcgtgcgc ccgtacgcgg 48780

tgaccgccag gccggcccgc gccgccaggg cacggaactc gtccagcgtg cggggcttgc 48840tgaccgccag gccggcccgc gccgccaggg cacggaactc gtccagcgtg cggggcttgc 48840

cgccggtctg caccatcatc atcagcccgg cccgggcggc ctcccgcccg ggggagacac 48900cgccggtctg caccatcatc atcagcccgg cccgggcggc ctcccgcccg ggggagacac 48900

caccgaccac gacgacgcga ccccgtgggg ctgcggcctc ggcgcaccgg gtgagcaggg 48960caccgaccac gacgacgcga ccccgtgggg ctgcggcctc ggcgcaccgg gtgagcaggg 48960

cggcggcctc ctcgtcgggc cagtccagca gcaggcgctt gagcaggtac acgtccgctc 49020cggcggcctc ctcgtcgggc cagtccagca gcaggcgctt gagcaggtac acgtccgctc 49020

cgggcgggag cgggtcgaag aagctctgcc cgctgaccgc ggcccggtcg gcgaccccgg 49080cgggcgggag cgggtcgaag aagctctgcc cgctgaccgc ggcccggtcg gcgaccccgg 49080

cggcggcgaa gacctccgcg gctcgggcga ccgcacgggg caggtcgacc agggtgccgt 49140cggcggcgaa gacctccgcg gctcgggcga ccgcacgggg caggtcgacc agggtgccgt 49140

gcacgtgcgg ccggacccgc aggatctcgg ccagcgtctc gccggtaccg ccgcccacgt 49200gcacgtgcgg ccggacccgc aggatctcgg ccagcgtctc gccggtaccg ccgcccacgt 49200

cgacgacgtg gtggacgtcc gcccagtcgt cgtcgacgag aacggcggga tccgaggcgt 49260cgacgacgtg gtggacgtcc gcccagtcgt cgtcgacgag aacggcggga tccgaggcgt 49260

cgccggcccg atggcccatc atggcgtcgt agctctccgc gatctcggga tgagcctcga 49320cgccggcccg atggcccatc atggcgtcgt agctctccgc gatctcggga tgagcctcga 49320

ggtcctccca gaacgagcgt ccgaagacgg tctcgtagcc cgggcggccg gtgcggacgg 49380ggtcctccca gaacgagcgt ccgaagacgg tctcgtagcc cgggcggccg gtgcggacgg 49380

tcgcgagcag cccgctccag gcggcggcgc tgcgaccgcc gtggccgtcg agggtgagcc 49440tcgcgagcag cccgctccag gcggcggcgc tgcgaccgcc gtggccgtcg agggtgagcc 49440

cgcgcagcgg accgggctcc agcagggtgc gggccgcctc gttcaacgcg aactcgccct 49500cgcgcagcgg accgggctcc agcagggtgc gggccgcctc gttcaacgcg aactcgccct 49500

cggccggctc ctcgaagacg cccttgccca ccagatggcg caggacccgc gcgagcgcgt 49560cggccggctc ctcgaagacg cccttgccca ccagatggcg caggacccgc gcgagcgcgt 49560

cgcgatcgca gtccgtcttc cgggcgagct ccccgatcgt cgtgacgccg gctgccacgt 49620cgcgatcgca gtccgtcttc cgggcgagct ccccgatcgt cgtgacgccg gctgccacgt 49620

gctcggcgat gcccagtgtc accaccaccc gcaggctcca cggggtcgcc agatcactga 49680gctcggcgat gcccagtgtc accaccaccc gcaggctcca cggggtcgcc agatcactga 49680

gcgaatcgac atccaccttc tccgacattt gccacctcca gtggtcgacg cggctcgaca 49740gcgaatcgac atccaccttc tccgacattt gccacctcca gtggtcgacg cggctcgaca 49740

gcccacggct gggccgcctg gaattcgctg ccggaatttc cccaccggcc ctggccgaag 49800gcccacggct gggccgcctg gaattcgctg ccggaatttc cccaccggcc ctggccgaag 49800

tcggcccact gccacgcctg cgggcgggcg gcaccggagc gcctcacgat agcggcattc 49860tcggcccact gccacgcctg cgggcgggcg gcaccggagc gcctcacgat agcggcattc 49860

atcgatccgc acagctcgtt cagaactgcg cgcgatggtt ccggatgccg ttcgaccggc 49920atcgatccgc acagctcgtt cagaactgcg cgcgatggtt ccggatgccg ttcgaccggc 49920

gagaacgttc agttgaacac ggccgaccgc aatcgcgagg ttgtggcggg atcggccaga 49980gagaacgttc agttgaacac ggccgaccgc aatcgcgagg ttgtggcggg atcggccaga 49980

tgccaagctc cgtcgaggtc gagtacgtcg gcgctcaccg agcgattaaa tcggcatttc 50040tgccaagctc cgtcgaggtc gagtacgtcg gcgctcaccg agcgattaaa tcggcatttc 50040

cggctcctct cctctggcga aagacgaatc gacatggcag acacgtacca agccagccgg 50100cggctcctct cctctggcga aagacgaatc gacatggcag acacgtacca agccagccgg 50100

caatgggaac gcatcagcag gcactgggtg accgaggaag cggccgtcga cctcgcgaat 50160caatgggaac gcatcagcag gcactgggtg accgaggaag cggccgtcga cctcgcgaat 50160

ttcaagtcgg ggcggccgaa ccacaagatc tcgctgtgga atcccgaggt gaacggcgtc 50220ttcaagtcgg ggcggccgaa ccacaagatc tcgctgtgga atcccgaggt gaacggcgtc 50220

cggtacctga agacgctcgt ccacaacctg gcgacagcgc tcgggccggc cgactgggag 50280cggtacctga agacgctcgt ccacaacctg gcgacagcgc tcgggccggc cgactgggag 50280

cggctgcgca ggaccaccca ccgggacgtg ggcaacccca tcgccgtccg cgtgggcggc 50340cggctgcgca ggaccaccca ccgggacgtg ggcaacccca tcgccgtccg cgtgggcggc 50340

gagagcatct gcctcgatta cctgcaggcc accctcgaac tccgtttcat cgagcagagc 50400gagagcatct gcctcgatta cctgcaggcc accctcgaac tccgtttcat cgagcagagc 50400

gtcgacctgg cgggtgcgag cgtcctcgag atcggtgcgg gctacggacg aacgtgtcac 50460gtcgacctgg cgggtgcgag cgtcctcgag atcggtgcgg gctacggacg aacgtgtcac 50460

accctgctgt cgaaccacga catcgccgcc tactgcatcg tggacctgcg cagcaccatg 50520accctgctgt cgaaccacga catcgccgcc tactgcatcg tggacctgcg cagcaccatg 50520

cacctcagcc ggcggtatct ccgggaggtc ctcgacgacg cccggttcgc caagatccgt 50580cacctcagcc ggcggtatct ccgggaggtc ctcgacgacg cccggttcgc caagatccgt 50580

ttcgtcccgg tggaggacga ggacgtgaac gcggcgctgc gcgagagcga gttcgatctg 50640ttcgtcccgg tggaggacga ggacgtgaac gcggcgctgc gcgagagcga gttcgatctg 50640

tgcatcaaca tcgactcgtt cgccgagatg gccccggaga ccgcacggag ctacctcgac 50700tgcatcaaca tcgactcgtt cgccgagatg gccccggaga ccgcacggag ctacctcgac 50700

ctcatcggcg agcggggcac ggccctgtac gtgaagaacc cggtcggcaa gtactgggac 50760ctcatcggcg agcggggcac ggccctgtac gtgaagaacc cggtcggcaa gtactgggac 50760

cagagcctgg acgggcacgt cgagggcgac gacgccctgc gccgggcgat ggagaccggc 50820cagagcctgg acgggcacgt cgagggcgac gacgccctgc gccgggcgat ggagaccggc 50820

ctgctgcgcg aggtgctcga catccacgac gaccgggcgg tgcgcgctgc cgtcccggag 50880ctgctgcgcg aggtgctcga catccacgac gaccgggcgg tgcgcgctgc cgtcccggag 50880

ttcatcgcgg cctaccggcc cggtgacgac tggcgctgcg tcgccgacgc ctgggccgtg 50940ttcatcgcgg cctaccggcc cggtgacgac tggcgctgcg tcgccgacgc ctgggccgtg 50940

ccgtggagct tctactggca ggcgatctat cgccggggga ggccgagcac cggcccgcag 51000ccgtggagct tctactggca ggcgatctat cgccggggga ggccgagcac cggcccgcag 51000

cggtgaccgt gccgggggac ggaagcaggc gagtaccgaa ggtcggagga gcggtgggtc 51060cggtgaccgt gccgggggac ggaagcaggc gagtaccgaa ggtcggagga gcggtgggtc 51060

cactggtgag cgtgctgggt gcgtccggat atctgggatc ggtcgtcacc gcccgcctct 51120cactggtgag cgtgctgggt gcgtccggat atctgggatc ggtcgtcacc gcccgcctct 51120

cgcagctccc gatccggctg cgtgccgtgt cgcgccgggt cagcccggtc ccggacgagg 51180cgcagctccc gatccggctg cgtgccgtgt cgcgccgggt cagcccggtc ccggacgagg 51180

tggtggccga cgtggaggtc cgtaccgccg acctcaccga cccgcgatgt ctcgccgacg 51240tggtggccga cgtggaggtc cgtaccgccg acctcaccga cccgcgatgt ctcgccgacg 51240

ccgtcgcgga cgccgacgtg atccttcacc tcggtaagca cagcggcgga tggcgtgacg 51300ccgtcgcgga cgccgacgtg atccttcacc tcggtaagca cagcggcgga tggcgtgacg 51300

cggacacgcc ggagggcgat cgggtgaacg tcggcgtcgc gcgcgacctc gtcgacatcc 51360cggacacgcc ggagggcgat cgggtgaacg tcggcgtcgc gcgcgacctc gtcgacatcc 51360

tcggccggcg acggccggca gcggcgcctc cggtggtggt gttcgccgcg acgacctcgc 51420tcggccggcg acggccggca gcggcgcctc cggtggtggt gttcgccgcg acgacctcgc 51420

aggtcggccg tccgcccgag caccctatga acggcagcga accggaccgc ccggagacgg 51480aggtcggccg tccgcccgag caccctatga acggcagcga accggaccgc ccggagacgg 51480

cgtacgaccg tcagaagctc cgggccgagg gcatcctcaa ggccgcgacc gaggcgtcgg 51540cgtacgaccg tcagaagctc cgggccgagg gcatcctcaa ggccgcgacc gaggcgtcgg 51540

tgatccgggg cgtgagcctg cggctgccga cggtcttcgg ccagagcggg ttgtcccgcg 51600tgatccgggg cgtgagcctg cggctgccga cggtcttcgg ccagagcggg ttgtcccgcg 51600

tacccgacgt cggcgtggtc tccgcgatgg cgcgtcaggc gatcgccggc cggccgttga 51660tacccgacgt cggcgtggtc tccgcgatgg cgcgtcaggc gatcgccggc cggccgttga 51660

ccatgtggca cgacggaacg gtcaagcgcg acctggtgta cgtcgaggac gcggcggacg 51720ccatgtggca cgacggaacg gtcaagcgcg acctggtgta cgtcgaggac gcggcggacg 51720

ccttcctggc agccatgcgg cgacctgacg aactggccgg ccggcactgg ctggtgggca 51780ccttcctggc agccatgcgg cgacctgacg aactggccgg ccggcactgg ctggtgggca 51780

ccggccggca cgacacggtc ggctcggtgt tccgtgtgat cgcggcgtcg gtcgccgcgc 51840ccggccggca cgacacggtc ggctcggtgt tccgtgtgat cgcggcgtcg gtcgccgcgc 51840

acacgggtcg gccgccggtg ccggtgacgt ccgtgccgcc gcccgcgcac gcgcctgtga 51900acacgggtcg gccgccggtg ccggtgacgt ccgtgccgcc gcccgcgcac gcgcctgtga 51900

tcgattttct gagcgtcacg attgatcccg cacccttcca gctcgcttcc ggctggcggg 51960tcgattttct gagcgtcacg attgatcccg cacccttcca gctcgcttcc ggctggcggg 51960

cgcggacggc gctcgacgat gccgtggacc gcactgtcgc cgctctgctg gatcgcggtg 52020cgcggacggc gctcgacgat gccgtggacc gcactgtcgc cgctctgctg gatcgcggtg 52020

aagctaccgg agaggagccc catgtacgga cgtgacgtgg ccgagatcca tgacgacctc 52080aagctaccgg agaggagccc catgtacgga cgtgacgtgg ccgagatcca tgacgacctc 52080

aacgagagcc ggggaaagga ctaccggacc gaagcggagt acatcaccga ggtggtccgt 52140aacgagagcc ggggaaagga ctaccggacc gaagcggagt acatcaccga ggtggtccgt 52140

agccggttcc ccggagcgcg ttccctgctc gacatcgggt gcggcgcggg cggccacctc 52200agccggttcc ccggagcgcg ttccctgctc gacatcgggt gcggcgcggg cggccacctc 52200

gtgcacttcg ccgagttctt cgacaccgtc ggagggatcg agctgtcgga ggacatgctg 52260gtgcacttcg ccgagttctt cgacaccgtc ggagggatcg agctgtcgga ggacatgctg 52260

gccgtggccc ggggcaagct gcccggtgcc gggctgcacc agggcgacat gcgggggttc 52320gccgtggccc ggggcaagct gcccggtgcc gggctgcacc agggcgacat gcgggggttc 52320

gacctcggcc gcgagtacga cgccgtggtg tgcctgttcg ggtccgtcgg ccacacccat 52380gacctcggcc gcgagtacga cgccgtggtg tgcctgttcg ggtccgtcgg ccacacccat 52380

gacgaggacg aactgcggca gacgctgcgc tgcttcgggc ggcatctctc cgccggtggc 52440gacgaggacg aactgcggca gacgctgcgc tgcttcgggc ggcatctctc cgccggtggc 52440

gtcgtcgtgg tcgagccgtg gtggttcctg gagaagtcgg tggacggatt catctccggt 52500gtcgtcgtgg tcgagccgtg gtggttcctg gagaagtcgg tggacggatt catctccggt 52500

gacgtcgtcc ggagcggccg gtcgaccatc gcgcggatgt cgcacaccgc gcgttccggg 52560gacgtcgtcc ggagcggccg gtcgaccatc gcgcggatgt cgcacaccgc gcgttccggg 52560

caccggtcga caatggacgt gcacttcctg gtggccgacc cggagaccgg ggtgcggcac 52620caccggtcga caatggacgt gcacttcctg gtggccgacc cggagaccgg ggtgcggcac 52620

ttcgccgaga cgtacaccca cacgttgttc agccgggccc agtacgaggc ggccttcgcc 52680ttcgccgaga cgtacaccca cacgttgttc agccgggccc agtacgaggc ggccttcgcc 52680

gcggccggct tcgacgccga ctacatcgag gacgtccagg gcggccgggg actcttcgtc 52740gcggccggct tcgacgccga ctacatcgag gacgtccagg gcggccgggg actcttcgtc 52740

gccgtcgccg tcgaggagcg gccgtgacga tccgggtgtg ggactacctc gccgagtacg 52800gccgtcgccg tcgaggagcg gccgtgacga tccgggtgtg ggactacctc gccgagtacg 52800

agagtgagcg tccggacgtc ctcgacgcgg tggagacggt gttccggtcc ggccagctgg 52860agagtgagcg tccggacgtc ctcgacgcgg tggagacggt gttccggtcc ggccagctgg 52860

tgctcggcgc gagcgtccag ggcttcgagg aggcgttcgc cgcctaccac ggcgtcccgc 52920tgctcggcgc gagcgtccag ggcttcgagg aggcgttcgc cgcctaccac ggcgtcccgc 52920

actgcgtcag cgtggacaac gggacgaacg ccatcaagct gggtctgcag gcgctcggcg 52980actgcgtcag cgtggacaac gggacgaacg ccatcaagct gggtctgcag gcgctcggcg 52980

tgggacccgg agacgaggtc gtcaccgtgt ccaacaccgc cgcaccgacc gtcgtggcga 53040tgggacccgg agacgaggtc gtcaccgtgt ccaacaccgc cgcaccgacc gtcgtggcga 53040

tcgacgcggt gggcgccacg ccggtgttcg tcgacatccg ggcggacgac tacctgatgg 53100tcgacgcggt gggcgccacg ccggtgttcg tcgacatccg ggcggacgac tacctgatgg 53100

acaccacgca ggtggaggcg gtcctcaccg aacggacccg ctgcctgctt ccggttcacc 53160acaccacgca ggtggaggcg gtcctcaccg aacggacccg ctgcctgctt ccggttcacc 53160

tgtacggaca gtgcgtcgac atgcggccgc tgcgcgaggt cgccgaccgc cacggcctgc 53220tgtacggaca gtgcgtcgac atgcggccgc tgcgcgaggt cgccgaccgc cacggcctgc 53220

tgatcctgga ggactgcgcc caggcacacg gggcacgcca cggcggtgtg ctcgccggca 53280tgatcctgga ggactgcgcc caggcacacg gggcacgcca cggcggtgtg ctcgccggca 53280

cgatgggcga cgccgcggcg ttctcgttct atccgacgaa ggtgctcggc gcgtacggtg 53340cgatgggcga cgccgcggcg ttctcgttct atccgacgaa ggtgctcggc gcgtacggtg 53340

acggtggcgc gacgatcacc gcggacgacg cggtggcggc caacctgcgg cgcctgcggt 53400acggtggcgc gacgatcacc gcggacgacg cggtggcggc caacctgcgg cgcctgcggt 53400

actacggcat ggagagccgc tactacgtcg tgcagacgcc ggggcacaac agccggctgg 53460actacggcat ggagagccgc tactacgtcg tgcagacgcc ggggcacaac agccggctgg 53460

acgaggtgca ggccgagatc ctgcgtcgca agctcacacg tctcgaccgg tacatcgcgg 53520acgaggtgca ggccgagatc ctgcgtcgca agctcacacg tctcgaccgg tacatcgcgg 53520

accggcgcgc ggtggcgcag cggtacgcgg agggcctcgg ggacaccgac ctggtcctgc 53580accggcgcgc ggtggcgcag cggtacgcgg agggcctcgg ggacaccgac ctggtcctgc 53580

cccgggtggc cgacgggaac gaccacgtgt actacctcta cgtggtgcgg cacccccgcc 53640cccgggtggc cgacgggaac gaccacgtgt actacctcta cgtggtgcgg cacccccgcc 53640

gggacgagat cctgacgcgg ctgcgcggct acgggatcga gctgaacgtc agctacccct 53700gggacgagat cctgacgcgg ctgcgcggct acgggatcga gctgaacgtc agctacccct 53700

ggccggtgca caccatgacc gggttcgccc acctcggcta ccgggagggc tcgcttccgg 53760ggccggtgca caccatgacc gggttcgccc acctcggcta ccgggagggc tcgcttccgg 53760

tcaccgaggc gctggccggg cagatcttct cactgccgat gtatccgtct ctcccggtcg 53820tcaccgaggc gctggccggg cagatcttct cactgccgat gtatccgtct ctcccggtcg 53820

acgtccagga gaagacgatc agtgccctcc gagacatcct caagacgctg tgaacggccg 53880acgtccagga gaagacgatc agtgccctcc gagacatcct caagacgctg tgaacggccg 53880

gttctaggga gatcaggtga aggtcgaaga gctcgcggtc accggtgcgt tcgtgttcac 53940gttctaggga gatcaggtga aggtcgaaga gctcgcggtc accggtgcgt tcgtgttcac 53940

gcccgacgtc taccccgacc atcgtggatc gttcgtgtcg ccgttccagc aacgggcctt 54000gcccgacgtc taccccgacc atcgtggatc gttcgtgtcg ccgttccagc aacgggcctt 54000

cgcctcggcg aagggtgcgc cgttcctccc cgtcgcgcag acgaatcaca gcgtctcccg 54060cgcctcggcg aagggtgcgc cgttcctccc cgtcgcgcag acgaatcaca gcgtctcccg 54060

ccggggcgtc gtgcggggca tccactacac cgtcaccccg ccgggcacga cgaagtacgt 54120ccggggcgtc gtgcggggca tccactacac cgtcaccccg ccgggcacga cgaagtacgt 54120

ctactgtgcg gcgggtgagg ccatcgacat cgtggtcgac atccgggtcg gctcgccgac 54180ctactgtgcg gcgggtgagg ccatcgacat cgtggtcgac atccgggtcg gctcgccgac 54180

gtacggcaag tgggacgccg tgcgggtgaa cccgcgggac ttccgcgcgg tgtacttccc 54240gtacggcaag tgggacgccg tgcgggtgaa cccgcgggac ttccgcgcgg tgtacttccc 54240

ggtcggggtc gggcacgcct tcgtggccct ggccgacgac acggtcatgt cgtacatgct 54300ggtcggggtc gggcacgcct tcgtggccct ggccgacgac acggtcatgt cgtacatgct 54300

gaccagcgcc tacgtgcccg agtacgaaaa ggcgatctcc gtgttcgacc cggacctggg 54360gaccagcgcc tacgtgcccg agtacgaaaa ggcgatctcc gtgttcgacc cggacctggg 54360

gctgccgatt cccggggaca tcgagccgat cgtctccgag cgggacagcg tgggcccgcg 54420gctgccgatt cccggggaca tcgagccgat cgtctccgag cgggacagcg tgggcccgcg 54420

gctggcggag gcggccgagg cgggcctgct accggactac caggagtgcc gggccatcga 54480gctggcggag gcggccgagg cgggcctgct accggactac caggagtgcc gggccatcga 54480

ggagcggctg ctccgctccg cctcgtgagc tgcgccggga cgtggcggaa ccgcacccgc 54540ggagcggctg ctccgctccg cctcgtgagc tgcgccggga cgtggcggaa ccgcacccgc 54540

gggccgggtc cgccgcacga cgagccggcc cggtgaccgg aggtccggtc gccggggtcc 54600gggccgggtc cgccgcacga cgagccggcc cggtgaccgg aggtccggtc gccggggtcc 54600

cggcgccgcc gcgcacccgt tcgccccgct cgctgcccac ccggttccgg tgaaaggacg 54660cggcgccgcc gcgcacccgt tcgccccgct cgctgcccac ccggttccgg tgaaaggacg 54660

caccgcatgc catctgtggt tcgcatggga gtcctggggt gcgcgagcat cgcgctccgc 54720caccgcatgc catctgtggt tcgcatggga gtcctggggt gcgcgagcat cgcgctccgc 54720

cgggttctgc cggcgatgat cgaggcggac ggcatggagc tgcgggccgt cgcgagccgc 54780cgggttctgc cggcgatgat cgaggcggac ggcatggagc tgcgggccgt cgcgagccgc 54780

gatcccgcca aggcgcacgc gatcgccgag cgtttcggct gcgtggccgc cgaggggtac 54840gatcccgcca aggcgcacgc gatcgccgag cgtttcggct gcgtggccgc cgaggggtac 54840

gacgacctgc tggaccggcc cgacatcgac gcggtctaca tcccgctgcc caccggcctg 54900gacgacctgc tggaccggcc cgacatcgac gcggtctaca tcccgctgcc caccggcctg 54900

cacgcgtact gggcgagccg cgccctcgcc gccggcaagc acgtgctctc ggagaagccg 54960cacgcgtact gggcgagccg cgccctcgcc gccggcaagc acgtgctctc ggagaagccg 54960

ctcaccagtg accacgccac cgcccgtgac ctcgtgaacc aggcgaaggg cgcgggtctg 55020ctcaccagtg accacgccac cgcccgtgac ctcgtgaacc aggcgaaggg cgcgggtctg 55020

tggctgatcg agaactacat gttcctccac caccgccagc acgacatggt ccgtgacctc 55080tggctgatcg agaactacat gttcctccac caccgccagc acgacatggt ccgtgacctc 55080

gtcgcgcagg gcaggatcgg cgagccccgc gtcttcaccg cgagcttcgg catccccccg 55140gtcgcgcagg gcaggatcgg cgagccccgc gtcttcaccg cgagcttcgg catccccccg 55140

ctcgacgcgg cgaactggcg gtaccacgtc gagctcggtg gtggggcgct gctcgacgtg 55200ctcgacgcgg cgaactggcg gtaccacgtc gagctcggtg gtggggcgct gctcgacgtg 55200

ggcgtctacc cactgcgcgc cgccacctac ttcctcggcg ccgacctgga ggtcgtcggg 55260ggcgtctacc cactgcgcgc cgccacctac ttcctcggcg ccgacctgga ggtcgtcggg 55260

tcggtgctgc gcatccatcc ggtccggggc gtcgacgtgg ccgggcacgc cctgctcagt 55320tcggtgctgc gcatccatcc ggtccggggc gtcgacgtgg ccgggcacgc cctgctcagt 55320

accccgtcgg gtgtgacggc cgagctgtcc ttcggtttcg agcacgcgta ccggtcctgc 55380accccgtcgg gtgtgacggc cgagctgtcc ttcggtttcg agcacgcgta ccggtcctgc 55380

tactcgctgt ggggcgaccg ggcccgtctc accctggacc gggccttcac accgccggtg 55440tactcgctgt ggggcgaccg ggcccgtctc accctggacc gggccttcac accgccggtg 55440

acccggcaac cagtggtccg gatcgaagcg gaggaccacg ccgaggaggt cgttctcccc 55500acccggcaac cagtggtccg gatcgaagcg gaggaccacg ccgaggaggt cgttctcccc 55500

gccgaccacc agttcaagaa catcgcggag ttcttcgccc ggtccctgct cgacggcggg 55560gccgaccacc agttcaagaa catcgcggag ttcttcgccc ggtccctgct cgacggcggg 55560

gactacacgc cgcacgccga agcgatctgc cagcaggccg agctcgtcga caaggtgcgg 55620gactacacgc cgcacgccga agcgatctgc cagcaggccg agctcgtcga caaggtgcgg 55620

tccagcgcga tccgcatcac cgagccccgc cgggtgcagt ggagcgggat cgccggatga 55680tccagcgcga tccgcatcac cgagccccgc cgggtgcagt ggagcgggat cgccggatga 55680

cagggccggc ggtcctcgac ggagaggaac cgccggcccg cgccggacgt ccggtcaccg 55740cagggccggc ggtcctcgac ggagaggaac cgccggcccg cgccggacgt ccggtcaccg 55740

ggacggtcgc gccccgccgg ttctcgacgc catgaactcc gcgaccgtcg ccacgaacgt 55800ggacggtcgc gccccgccgg ttctcgacgc catgaactcc gcgaccgtcg ccacgaacgt 55800

cgccttgtcg gcgtccggca ggcggaagta cggcgcgagg acctcggaca accccggccg 55860cgccttgtcg gcgtccggca ggcggaagta cggcgcgagg acctcggaca accccggccg 55860

gacgaacgtc atcggctggt cgcgccggtg cagcggttcg atcacgcaca cgtccccggg 55920gacgaacgtc atcggctggt cgcgccggtg cagcggttcg atcacgcaca cgtccccggg 55920

cccggtgccg cggtacgcca tgtccgtggc gaccgtgaac cactgctgtt ccccggcgat 55980cccggtgccg cggtacgcca tgtccgtggc gaccgtgaac cactgctgtt ccccggcgat 55980

cgttccgcgc gcgcgggcca ccgactgttc cttcaggcag cgcggagcat gactggcggc 56040cgttccgcgc gcgcgggcca ccgactgttc cttcaggcag cgcggagcat gactggcggc 56040

ggcgatctgc agcaccgtgt cgacgccgtg cgcggcgaag atgccggcgg ccgcctcgat 56100ggcgatctgc agcaccgtgt cgacgccgtg cgcggcgaag atgccggcgg ccgcctcgat 56100

ctcggcgacc gtgttgtgca ccggctgcgt gatgacgatg tcgtgcaggg cctggcggaa 56160ctcggcgacc gtgttgtgca ccggctgcgt gatgacgatg tcgtgcaggg cctggcggaa 56160

cgccgcgcgc tccgcgtccg acaaccggtc gagcaggggc ttcaggcggc tgaactcgtc 56220cgccgcgcgc tccgcgtccg acaaccggtc gagcaggggc ttcaggcggc tgaactcgtc 56220

cagcctggag aggttgtcca gaaagaacct cttggtgtac tcgccctcac cgacgccgtc 56280cagcctggag aggttgtcca gaaagaacct cttggtgtac tcgccctcac cgacgccgtc 56280

ccgccgggag aagccgccgc cgatgacgat gcaggcgagt tcgtgatcgc cgcccagtgt 56340ccgccgggag aagccgccgc cgatgacgat gcaggcgagt tcgtgatcgc cgcccagtgt 56340

caggacggct cgcgccagcg tgcccaggtc acccagacgg tcgtcctcgg gcacgccgac 56400caggacggct cgcgccagcg tgcccaggtc acccagacgg tcgtcctcgg gcacgccgac 56400

gaccagttcc gcccacccgt cggtggccag atggcggctg tgcaggagca ctcccactct 56460gaccagttcc gcccacccgt cggtggccag atggcggctg tgcaggagca ctcccactct 56460

cgtcgtcaac acggaactcc tcccggtggg ggcacgacgg aggctcgccc gatccgggcg 56520cgtcgtcaac acggaactcc tcccggtggg ggcacgacgg aggctcgccc gatccgggcg 56520

gcgagaaggg tgagggccgg cggctagccg gcggcggcca gctgctcgat cgtgccgacc 56580gcgagaaggg tgagggccgg cggctagccg gcggcggcca gctgctcgat cgtgccgacc 56580

atctcggccg tcgtgggcag cttaccgatc tcggtggcca ggacccgcgc gcgctcggcg 56640atctcggccg tcgtgggcag cttaccgatc tcggtggcca ggacccgcgc gcgctcggcg 56640

aaccggcggt ccgagaggat ctgccggcag gccgcggcca cggcgtcgac cgcttcccgg 56700aaccggcggt ccgagaggat ctgccggcag gccgcggcca cggcgtcgac cgcttcccgg 56700

tacgcctcct ggccggcctg gcggggggcc ggcggcacgg cgagggcggc accgaagtcg 56760tacgcctcct ggccggcctg gcggggggcc ggcggcacgg cgagggcggc accgaagtcg 56760

cccagggcct tcgcgatggt cctcgagtag tcgttgtccg gggtgatcag ctgcggcacg 56820cccaggggcct tcgcgatggt cctcgagtag tcgttgtccg gggtgatcag ctgcggcacg 56820

ccggcgttca tgagcgtcat cgcggtggcc gcgccgccgt ggtgcaccgc gaggtcgcag 56880ccggcgttca tgagcgtcat cgcggtggcc gcgccgccgt ggtgcaccgc gaggtcgcag 56880

gtgggagcga cgacgtcgag cggcacccag ccgatgcgca cgtcggagaa ctgtgcgccg 56940gtgggagcga cgacgtcgag cggcacccag ccgatgcgca cgtcggagaa ctgtgcgccg 56940

aactcctcgg cggcgcggtc cggtgccgcg atcacgacct ccgccccggc tgccgtcagc 57000aactcctcgg cggcgcggtc cggtgccgcg atcacgacct ccgccccggc tgccgtcagc 57000

tggtcgacca gctgacgcat gacgccggtg tgcgagtgca gcatgagatt gcgcgttccc 57060tggtcgacca gctgacgcat gacgccggtg tgcgagtgca gcatgagatt gcgcgttccc 57060

gcggtgacca gcactcgtgg gcgccctgcg ggccgggtgt acatccacgg gtcgagtcgg 57120gcggtgacca gcactcgtgg gcgccctgcg ggccgggtgt acatccacgg gtcgagtcgg 57120

ccctggcggt tcctcgggat ccagcgcatc ggtcgtgcgc ccggggtggg cgacggccgc 57180ccctggcggt tcctcgggat ccagcgcatc ggtcgtgcgc ccggggtggg cgacggccgc 57180

aggcacggcg ggcagacgtc gatgaacagg tcggcctccg gcggcccggt gaggccgaga 57240aggcacggcg ggcagacgtc gatgaacagg tcggcctccg gcggcccggt gaggccgaga 57240

cgctccaggt cgggctcgat tcccggctcc ggccgcagcg ggacgatgtc ccagtagtgg 57300cgctccaggt cgggctcgat tcccggctcc ggccgcagcg ggacgatgtc ccagtagtgg 57300

cgtacgtaca gcgccttcag atgggtggcg aggatgccgg ggacgtgcga caggccaccg 57360cgtacgtaca gcgccttcag atgggtggcg aggatgccgg ggacgtgcga caggccaccg 57360

acgaccacat cggccggcca ctgccgggtc agctccagca gcgcctgcag cctggccgcg 57420acgaccacat cggccggcca ctgccgggtc agctccagca gcgcctgcag cctggccgcg 57420

ggcatccccg gatgaggtga ggggaccacc ggcagtccga tcgccgtcgc cgtctccagc 57480ggcatccccg gatgaggtga ggggaccacc ggcagtccga tcgccgtcgc cgtctccagc 57480

agcggctcgt cggcggtcac gaggatctcg tgcccggcgc tccgggccgc cgtcgcgagc 57540agcggctcgt cggcggtcac gaggatctcg tgcccggcgc tccgggccgc cgtcgcgagc 57540

ggggcgatgg agaacacggt ggactggcta ccgcctacgg tgaacaggaa tttcatcgtg 57600ggggcgatgg agaacacggt ggactggcta ccgcctacgg tgaacaggaa tttcatcgtg 57600

atccgtcttc ttctcgacga ctgcggacag gtgcaccacg acggaccggc gtcctcgcgg 57660atccgtcttc ttctcgacga ctgcggacag gtgcaccacg acggaccggc gtcctcgcgg 57660

cgcgttcggg cacgtgcggg ggtccacgcg agacgcgcgg gcggcggcgg accggcccgg 57720cgcgttcggg cacgtgcggg ggtccacgcg agacgcgcgg gcggcggcgg accggcccgg 57720

gagccgcggc cgccggacga cgcgcaggcg gggccggggc gacagcgcgg cggccggggc 57780gagccgcggc cgccggacga cgcgcaggcg gggccggggc gacagcgcgg cggccggggc 57780

cggacgcctc acgtcccgct cccggatgtg gagggcggag ccgcgcggcg acgcagccgg 57840cggacgcctc acgtcccgct cccggatgtg gagggcggag ccgcgcggcg acgcagccgg 57840

tcggtgagtt ccggcccgtg ggagatcgcg agccgcacga tgtgacagat ccggtcgatg 57900tcggtgagtt ccggcccgtg ggagatcgcg agccgcacga tgtgacagat ccggtcgatg 57900

tccgtggggg agacggccgg gccggtcggc agggcgacca cctgctcggc caggcgctcg 57960tccgtggggg agacggccgg gccggtcggc agggcgacca cctgctcggc caggcgctcg 57960

gtgtgcggca gcgaggtcgg acgctccgag cggtagggct cgagctggtg gcaggcgggc 58020gtgtgcggca gcgaggtcgg acgctccgag cggtagggct cgagctggtg gcaggcgggc 58020

gagaagtagc gccgcgcccc ggcgttctcc gccgtcagca cctcgatcat caggtcgcgg 58080gagaagtagc gccgcgcccc ggcgttctcc gccgtcagca cctcgatcat caggtcgcgg 58080

tggatgccgg tgaccgcttc gtcgatcttg aggatcacgt actgccagtt gggatcctcc 58140tggatgccgg tgaccgcttc gtcgatcttg aggatcacgt actgccagtt gggatcctcc 58140

tgctcgtcga agtcgactac ggagagcccg cccaccccgc gcaacgcgga gcggtagtgc 58200tgctcgtcga agtcgactac ggagagcccg cccaccccgc gcaacgcgga gcggtagtgc 58200

gcgtggttgg cgcgattgtg ccgtacggtc tcgccgaagg cgtccagcga ggtcagtccc 58260gcgtggttgg cgcgattgtg ccgtacggtc tcgccgaagg cgtccagcga ggtcagtccc 58260

atggccgcgg aggcctcggt catcttggcg ttggtgccga tgcccacgct gcggccgtcg 58320atggccgcgg aggcctcggt catcttggcg ttggtgccga tgcccacgct gcggccgtcg 58320

gcggtgatgc cgaaggtccg caggacgcgg agctcctggg ccagcccgtc gtcgtcggtg 58380gcggtgatgc cgaaggtccg caggacgcgg agctcctggg ccagcccgtc gtcgtcggtg 58380

acgacggcgc cgccctcgaa gcagttgacc accttcgtgg cgtggaagct gaacacctcg 58440acgacggcgc cgccctcgaa gcagttgacc accttcgtgg cgtggaagct gaacacctcg 58440

gcggagccga agccgcccac gcgccggccc tggaagctgg accccagcgc gtgtgccgcg 58500gcggagccga agccgcccac gcgccggccc tggaagctgg accccagcgc gtgtgccgcg 58500

tcgaaggtga gcgtgaggcc gtgccgcgcg gccaccttct ccagctcgtc ggcccggccg 58560tcgaaggtga gcgtgaggcc gtgccgcgcg gccaccttct ccagctcgtc ggcccggccg 58560

gggcggcccc acaggtggac gccgacgatg ccggaggtcc gcggcgtgac ggccgcctcc 58620gggcggcccc acaggtggac gccgacgatg ccggaggtcc gcggcgtgac ggccgcctcc 58620

acctgctccg ggtcgaggca tccggtgacc ggatcgacgt cgcagaagac cggtgtcagg 58680acctgctccg ggtcgaggca tccggtgacc ggatcgacgt cgcagaagac cggtgtcagg 58680

cccagcaccc gggcggcgtg cggcgtggcc acataggtca tcgcgggcat gacgacctca 58740cccagcaccc gggcggcgtg cggcgtggcc acataggtca tcgcgggcat gacgacctca 58740

cccgacagcc cggtggccat gtagagcagt tgcagcgcca gggtgccgtt gcacgtcgcc 58800cccgacagcc cggtggccat gtagagcagt tgcagcgcca gggtgccgtt gcacgtcgcc 58800

acgcagtgcc ggacgcccgc caggtccgcg atgcgctcct cgaactcgcg gaccagcgcg 58860acgcagtgcc ggacgcccgc caggtccgcg atgcgctcct cgaactcgcg gaccagcgcg 58860

ccgttggtga gccattcgtt gtccatcgcc gtgttcagcc tgtcgaacag acgcgaccga 58920ccgttggtga gccattcgtt gtccatcgcc gtgttcagcc tgtcgaacag acgcgaccga 58920

tcgccgacgc tgggcgttcc cacctgcagg aaccgtgaga actccggggt cccgccgagc 58980tcgccgacgc tgggcgttcc cacctgcagg aaccgtgaga actccggggt cccgccgagc 58980

gcagcgagat cagcgagtgt gtctttcacg cgcgggcctc catcatgccg tgcgacgtgc 59040gcagcgagat cagcgagtgt gtctttcacg cgcgggcctc catcatgccg tgcgacgtgc 59040

catgggccgg cggtgtgccg gaacgacagc gtcccggacg gtgctcgcac ctcggttgag 59100catgggccgg cggtgtgccg gaacgacagc gtcccggacg gtgctcgcac ctcggttgag 59100

cgcaactggg gtgacgcccg ccggggctat caggcggaac cgctccaccg cccttccagt 59160cgcaactggg gtgacgcccg ccggggctat caggcggaac cgctccaccg cccttccagt 59160

ggacctccac catgggtcgg ccgtgcccgg cgacgctcgg aaacatgaaa cttctcgtga 59220ggacctccac catgggtcgg ccgtgcccgg cgacgctcgg aaacatgaaa cttctcgtga 59220

ccgggggagc cggcttcgtc ggctccgagt acgtacgcag catgctcggc ggcgcctacg 59280ccgggggagc cggcttcgtc ggctccgagt acgtacgcag catgctcggc ggcgcctacg 59280

agggttacga gaacgcggag atcaccgtcc tcgacaagct cacgtacgcg ggcagtctga 59340agggttacga gaacgcggag atcaccgtcc tcgacaagct cacgtacgcg ggcagtctga 59340

cgaacatccc ggtggacgat ccgcggctca cctttgtccg aggcgacatc gtcgaccgcg 59400cgaacatccc ggtggacgat ccgcggctca cctttgtccg aggcgacatc gtcgaccgcg 59400

agctgcttct cgacctgctg ccgggacacg acgcggtcgt ccacttcgcg gccgagagcc 59460agctgcttct cgacctgctg ccgggacacg acgcggtcgt ccacttcgcg gccgagagcc 59460

acgtggaccg gtcgctgctc gacgcctcgc cgttcacgac gacgaatgtg ctggggaccc 59520acgtggaccg gtcgctgctc gacgcctcgc cgttcacgac gacgaatgtg ctggggaccc 59520

agacgctgct ggactgctgc ctccggacag gcatcagccg ggtggtgcag gtctcgaccg 59580agacgctgct ggactgctgc ctccggacag gcatcagccg ggtggtgcag gtctcgaccg 59580

acgaggtgta cggcaccatc gcccacgggt cgtggacgga ggatcacccg ctcctgccga 59640acgaggtgta cggcaccatc gcccacgggt cgtggacgga ggatcacccg ctcctgccga 59640

attccccgta cgccgcctcg aaagcggcgg ccgacctgct cgcgcgctcg taccaccgct 59700attccccgta cgccgcctcg aaagcggcgg ccgacctgct cgcgcgctcg taccaccgct 59700

cgcacgggct gccggtggtg atcacccgct gctcgaacaa ctacgggccg taccagcacg 59760cgcacgggct gccggtggtg atcacccgct gctcgaacaa ctacgggccg taccagcacg 59760

tcgagaagat gatcccgcgc ttcgtcacca acctgctcag cggccggccg gtgcccctct 59820tcgagaagat gatcccgcgc ttcgtcacca acctgctcag cggccggccg gtgcccctct 59820

acggcgacgg ccgcaacgtc cgggagtggc tgcacgtcgc ggaccactgc cggggcgtgc 59880acggcgacgg ccgcaacgtc cgggagtggc tgcacgtcgc ggaccactgc cggggcgtgc 59880

agttggcgct ggggaagggt cgtgacggcg aggtctacca cctgggcagc ggcaccgagc 59940agttggcgct ggggaagggt cgtgacggcg aggtctacca cctgggcagc ggcaccgagc 59940

tgaccaaccg ggacctgacc gccgagctcc tgcgcctgtg cggggcggac tgggacgcgg 60000tgaccaaccg ggacctgacc gccgagctcc tgcgcctgtg cggggcggac tgggacgcgg 60000

tgcggccggt cgccgaccgc aaggggcacg acctccggta ctccctcgac gacagcaagg 60060tgcggccggt cgccgaccgc aaggggcacg acctccggta ctccctcgac gacagcaagg 60060

cccgccgcga gctcggctac gccccgcagg tgcccttcga gtcgggcctc gccgaggtcg 60120cccgccgcga gctcggctac gccccgcagg tgcccttcga gtcgggcctc gccgaggtcg 60120

tcgcctggta ccggaccaac agcgaccggt gggccgacga gtcggagcgg caccacacgc 60180tcgcctggta ccggaccaac agcgaccggt gggccgacga gtcggagcgg caccacacgc 60180

aagccgcacc agagccggca atgcccggtc cggacacaga ggaacagggg tcgatgaaca 60240aagccgcacc agagccggca atgcccggtc cggacacaga ggaacagggg tcgatgaaca 60240

tgaaggctgt cgggacgacc gcggcggtgt cgcggtgagt gttcgcaaag tggtgatcac 60300tgaaggctgt cgggacgacc gcggcggtgt cgcggtgagt gttcgcaaag tggtgatcac 60300

cggtctcggc gtcgtggcgc ccggcggcgt gggtaccaag gcgttctggc aactgatcac 60360cggtctcggc gtcgtggcgc ccggcggcgt gggtaccaag gcgttctggc aactgatcac 60360

cgccggtcgc accgcgacca ggccgatcac cgccttcgac gcgtccgcct tccggtcccg 60420cgccggtcgc accgcgacca ggccgatcac cgccttcgac gcgtccgcct tccggtcccg 60420

gatcgccgcc gaggtggact tcgaaccggc ccaggccgag ctgtcccacc gggagatcag 60480gatcgccgcc gaggtggact tcgaaccggc ccaggccgag ctgtcccacc gggagatcag 60480

ccggctggac cgggcggcgc agttcgccct ggtcgccacc agggaggcga tggccgacag 60540ccggctggac cgggcggcgc agttcgccct ggtcgccacc agggaggcga tggccgacag 60540

cggtctggag acggaccggt ccgatcccac ccgcgtcggc gtgagcctcg gcaccgccgt 60600cggtctggag acggaccggt ccgatcccac ccgcgtcggc gtgagcctcg gcaccgccgt 60600

gggtgccacg tgcaacctcg agtcggagta cctcgctctc agcgacaccg ggcgggagtg 60660gggtgccacg tgcaacctcg agtcggagta cctcgctctc agcgacaccg ggcgggagtg 60660

ggtgctcgac caccactacg ccggcccgca cctgtacgac tacttcatgc ccggctccat 60720ggtgctcgac caccactacg ccggcccgca cctgtacgac tacttcatgc ccggctccat 60720

tgcggcggag gtcgcctggg acgtcaacgc ccagggcccg gtcgccgtga tctccgcggg 60780tgcggcggag gtcgcctggg acgtcaacgc ccagggcccg gtcgccgtga tctccgcggg 60780

gtgcacctcc ggactggacg cggtcggaca cgcggtggat ctgatccggg agggcgcggt 60840gtgcacctcc ggactggacg cggtcggaca cgcggtggat ctgatccggg agggcgcggt 60840

cgacgcgatg gtgaccggtg gcaccgacgc gccgatctca ccgatcaccg tcgcctgctt 60900cgacgcgatg gtgaccggtg gcaccgacgc gccgatctca ccgatcaccg tcgcctgctt 60900

cgacgccatc cgggcgacct cgtcgagcaa cgacgacgcg gcccacgcgc tccgcccctt 60960cgacgccatc cgggcgacct cgtcgagcaa cgacgacgcg gcccacgcgc tccgcccctt 60960

tgaccgcacc cgcaacggct tcgtgctggg ggagggggcc gccgtgctcg tcctcgagag 61020tgaccgcacc cgcaacggct tcgtgctggg ggagggggcc gccgtgctcg tcctcgagag 61020

cgaggagcac gcccgcgccc gcggcgcccg gatctacgcc gaggtcaccg gattcgcctc 61080cgaggagcac gcccgcgccc gcggcgcccg gatctacgcc gaggtcaccg gattcgcctc 61080

ccgcagcaac gcgtaccaca tgaccggcct gcggcccgac ggcgccgaga tggccgcggc 61140ccgcagcaac gcgtaccaca tgaccggcct gcggcccgac ggcgccgaga tggccgcggc 61140

gatcaccgcc gcgctggcgg agagcaagct gtcaccggag gacgtcgact acgtcaacgc 61200gatcaccgcc gcgctggcgg agagcaagct gtcaccggag gacgtcgact acgtcaacgc 61200

gcacggcacg gcgacgcagc agaacgacag gcacgagacg gccgctctca agcgggcact 61260gcacggcacg gcgacgcagc agaacgacag gcacgagacg gccgctctca agcgggcact 61260

cggccaccac gcctactcga caccggtcag ttccatcaag tcgatggtgg gccactccct 61320cggccaccac gcctactcga caccggtcag ttccatcaag tcgatggtgg gccactccct 61320

cggtgcgatc ggctcgatcg agatcgccgc ctgcgcgctc gccctggacc agggcgtgat 61380cggtgcgatc ggctcgatcg agatcgccgc ctgcgcgctc gccctggacc agggcgtgat 61380

cccgccgacc gcgaacctcc acgagccgga ccccgagctg gacctggact acgtcccgct 61440cccgccgacc gcgaacctcc acgagccgga ccccgagctg gacctggact acgtcccgct 61440

gcacgcgcgg gagcagaggc tcgacaccgt cgtcagcgtc ggcagcggct tcggaggctt 61500gcacgcgcgg gagcagaggc tcgacaccgt cgtcagcgtc ggcagcggct tcggaggctt 61500

ccagagcgcc atcgtgctcg cccgtcccgg gcggggtgcg gcatgaccgc ggcggtgatc 61560ccagagcgcc atcgtgctcg cccgtcccgg gcggggtgcg gcatgaccgc ggcggtgatc 61560

accgggatcg gcgtcgcggc gccgaacggc ttcgggaccg agaacttctg ggccgcgacg 61620accgggatcg gcgtcgcggc gccgaacggc ttcgggaccg agaacttctg ggccgcgacg 61620

ttgcgtggcg agtccgccat ccgcccggtc cggcgattcg acgtgagcgg gtacccggcc 61680ttgcgtggcg agtccgccat ccgcccggtc cggcgattcg acgtgagcgg gtacccggcc 61680

cggctgggcg gcgaactcga cggattcgat cccgccgacc acattcccag ccggcttctc 61740cggctgggcg gcgaactcga cggattcgat cccgccgacc acattcccag ccggcttctc 61740

ccgcagaccg accggatgac ccagttcgcg ctggccgcgt cggcctgggc gctcgccgac 61800ccgcagaccg accggatgac ccagttcgcg ctggccgcgt cggcctgggc gctcgccgac 61800

gcccgggtgg aacccggcgc ctacgaggcg accgagaccg gcgtggcgat ggccggggcg 61860gcccgggtgg aacccggcgc ctacgaggcg accgagaccg gcgtggcgat ggccggggcg 61860

ttcggcggct tcgagtacgg ccagcgcgag ctggagaacc tgtggcggtc cggtcccgag 61920ttcggcggct tcgagtacgg ccagcgcgag ctggagaacc tgtggcggtc cggtcccgag 61920

cacgtcagcg tgtacatgtc gttcgcctgg ttctacgccg tcaacacggg gcagacgtcg 61980cacgtcagcg tgtacatgtc gttcgcctgg ttctacgccg tcaacacggg gcagacgtcg 61980

atccggcacg gcatgagagg ccccgccggc gtcatcgtca gcgaccaggc gggcgggctc 62040atccggcacg gcatgagagg ccccgccggc gtcatcgtca gcgaccaggc gggcgggctc 62040

gacgcggtcg cgcaggcccg gcggaacatc cgcaaggggt cccggctcat gctctccggg 62100gacgcggtcg cgcaggcccg gcggaacatc cgcaaggggt cccggctcat gctctccggg 62100

ggcttcgaga gctccttctg cccgtacggc tgggtggccc ggatgagcgg cggcacgctg 62160ggcttcgaga gctccttctg cccgtacggc tgggtggccc ggatgagcgg cggcacgctg 62160

agcaccggcg acgatccgcg aaccgcgtac gtgcccttcg acgtcggcgc ccgtgggtac 62220agcaccggcg acgatccgcg aaccgcgtac gtgcccttcg acgtcggcgc ccgtgggtac 62220

gtccccggtg agggtggtgc cgtcctggtg gtcgaggacg ccgccggcgc ccgccgccgg 62280gtccccggtg agggtggtgc cgtcctggtg gtcgaggacg ccgccggcgc ccgccgccgg 62280

ggcgcgcaca tccacgggga gatcgccggg tacgcggcga cgttcgacgc cgccgcgcgc 62340ggcgcgcaca tccacgggga gatcgccggg tacgcggcga cgttcgacgc cgccgcgcgc 62340

cacggtggtg cgcggggact gcgccgcgcc gtggaaggcg ccctggcgga cgcgcagatg 62400cacggtggtg cgcggggact gcgccgcgcc gtggaaggcg ccctggcgga cgcgcagatg 62400

acggccgcgg acatcggcgt ggtcttcgcc gacggctccg gaacgccgga cgaggaccac 62460acggccgcgg acatcggcgt ggtcttcgcc gacggctccg gaacgccgga cgaggaccac 62460

gccgaggccg aggcgatcgc cgccgtgttc ggtcccggcg cggttcccgt caccgtcccg 62520gccgaggccg aggcgatcgc cgccgtgttc ggtcccggcg cggttcccgt caccgtcccg 62520

aagaccatga ccggacggat gagctccggc ggcgcgtccg ccgacctcgc gtgcgccctg 62580aagaccatga ccggacggat gagctccggc ggcgcgtccg ccgacctcgc gtgcgccctg 62580

ctcgccatgc gcgacgggct cattccgccg acggtcaacg tgcgcacggt cgcggccggc 62640ctcgccatgc gcgacgggct cattccgccg acggtcaacg tgcgcacggt cgcggccggc 62640

gcgcagatcg acctcgtcac cggcgggccc cgccggtggg aacccgaggc cgcgctggtg 62700gcgcagatcg acctcgtcac cggcgggccc cgccggtggg aacccgaggc cgcgctggtg 62700

atcgcccggg gcaggggcgg attcaactcc gcgatggtgc tccgccgcgg caccgcgtcc 62760atcgcccggg gcaggggcgg attcaactcc gcgatggtgc tccgccgcgg caccgcgtcc 62760

ccggacgtca ccgcggaagc gacgagcaac gacagtgaca ttcgaaccgg atcaggaggg 62820ccggacgtca ccgcggaagc gacgagcaac gacagtgaca ttcgaaccgg atcaggaggg 62820

gagcagtcgt gcagcggata actctgccgg accttgagga gatcatgcgc aggtgcgccg 62880gagcagtcgt gcagcggata actctgccgg accttgagga gatcatgcgc aggtgcgccg 62880

gggacgacga gtcgacgtcc tcgttccagc aggctccgga ccaggcgttc accgacctcg 62940gggacgacga gtcgacgtcc tcgttccagc aggctccgga ccaggcgttc accgacctcg 62940

gatacgactc gctcgcgttg ctcgagacgc agagtgtcat caagcgggac tacggcgtcg 63000gatacgactc gctcgcgttg ctcgagacgc agagtgtcat caagcgggac tacggcgtcg 63000

agatctccga gcaggccctg agcgaggcca ccacgccgcg gcagctggtg gacctcgtga 63060agatctccga gcaggccctg agcgaggcca ccacgccgcg gcagctggtg gacctcgtga 63060

accgatggct gaccgcggcc tgaccggcgc cggggcgccg gccggccggg acacgccggc 63120accgatggct gaccgcggcc tgaccggcgc cggggcgccg gccggccggg acacgccggc 63120

cgagggcccg ccgcaaccgc cacctgccct gctgatgccg ggccagggtt cccagtacca 63180cgagggcccg ccgcaaccgc cacctgccct gctgatgccg ggccagggtt cccagtacca 63180

gggcatgggc acggggctgt accgggatgt ctccgccttc gcggcgatca tcgacgaggt 63240gggcatgggc acggggctgt accgggatgt ctccgccttc gcggcgatca tcgacgaggt 63240

gttcgagctc atggggaggg ccggggagca gctacgttcc gactggctcg ccgcctcccc 63300gttcgagctc atggggaggg ccggggagca gctacgttcc gactggctcg ccgcctcccc 63300

gcagcttccg gtcgaccatg ccagccgttc gcagccgctg ctgttcgcca tcgactacgc 63360gcagcttccg gtcgaccatg ccagccgttc gcagccgctg ctgttcgcca tcgactacgc 63360

gctcggcaga ctgctgctgg accgcgggct gcggccggcg gtcctgctgg gacacagtgt 63420gctcggcaga ctgctgctgg accgcgggct gcggccggcg gtcctgctgg gacacagtgt 63420

cggcgagatg gccgccgcca ccctcgccgg catcttcgac ctgcccggcg cgacccgcat 63480cggcgagatg gccgccgcca ccctcgccgg catcttcgac ctgcccggcg cgacccgcat 63480

cgtcgggcag cgggtcggcc agttcaccct ggtgccgccg ggcggaatgg ccgccgtggc 63540cgtcgggcag cgggtcggcc agttcaccct ggtgccgccg ggcggaatgg ccgccgtggc 63540

ggcgtcccgg gccgaggtcg agccgtacct gcggccgggt gtcgacgtcg gggcggtcaa 63600ggcgtcccgg gccgaggtcg agccgtacct gcggccgggt gtcgacgtcg gggcggtcaa 63600

cacgccccgg cagacggtga tcgccggtgc ggacgagccg ctgcggacca cagtggacgc 63660cacgccccgg cagacggtga tcgccggtgc ggacgagccg ctgcggacca cagtggacgc 63660

gctgcgccag gccggctaca cctgtgctcc ggtgccgtcg acggttccct tccacagcga 63720gctgcgccag gccggctaca cctgtgctcc ggtgccgtcg acggttccct tccacagcga 63720

gtggctccgg ccggcggtcg ccccggcctg ctccctgctg gcgagcctgc cggcgaaccc 63780gtggctccgg ccggcggtcg ccccggcctg ctccctgctg gcgagcctgc cggcgaaccc 63780

gccccggatc cccgtggtgt ccgggtacac cgccggctac ctgaccgagg ccgaggtgaa 63840gccccggatc cccgtggtgt ccgggtacac cgccggctac ctgaccgagg ccgaggtgaa 63840

ggatccacgg tactgggccg agcacccggt gaacccggtg ctgttctggc ccgccctggc 63900ggatccacgg tactgggccg agcacccggt gaacccggtg ctgttctggc ccgccctggc 63900

gagactggcg gaggccgggc cccacctgct ggtcgagtgc ggccccggca ccagcctgac 63960gagactggcg gaggccgggc cccacctgct ggtcgagtgc ggccccggca ccagcctgac 63960

caccttcgcc cgccggcacc cggacgtccg ttccgggcgg tgcgaggtgc tgtcgctgat 64020caccttcgcc cgccggcacc cggacgtccg ttccgggcgg tgcgaggtgc tgtcgctgat 64020

cgggccggcg gcgtcgggcc cggcccggga ggccgagcac ctcgctgccg cggcggcccg 64080cgggccggcg gcgtcgggcc cggcccggga ggccgagcac ctcgctgccg cggcggcccg 64080

gctgggagtc tcgctgccct gatccgcggt gccgccccgg gtcaaccgct gctcgaccgg 64140gctgggagtc tcgctgccct gatccgcggt gccgccccgg gtcaaccgct gctcgaccgg 64140

acctggtgag gccggtgcca ggctgcgcaa gcacgatcca cccaggcggt ggccgtcccc 64200acctggtgag gccggtgcca ggctgcgcaa gcacgatcca cccaggcggt ggccgtcccc 64200

taggaccgcg gagatgacat ggagaccaac cagcaggccc tgtttcccca ggtcggtcgt 64260taggaccgcg gagatgacat ggagaccaac cagcaggccc tgtttcccca ggtcggtcgt 64260

ccggcgctgg ttcgacgccc gggtgactgg ctcgaacgcg tcgaactgat cacccccgcg 64320ccggcgctgg ttcgacgccc gggtgactgg ctcgaacgcg tcgaactgat cacccccgcg 64320

atgtgcggcc ccaactcgct gttcatcggc cagctgggcg actggacctg ggacgcggtc 64380atgtgcggcc ccaactcgct gttcatcggc cagctgggcg actggacctg ggacgcggtc 64380

ggcgccgcct gccacatcga tccgtactcc gcggtggatc ccgagggcaa ccccgtctac 64440ggcgccgcct gccacatcga tccgtactcc gcggtggatc ccgagggcaa ccccgtctac 64440

ctgtccttcg cctatttccg ggtcaaggca tgcagcgacc tgtccgtcga gcaactgacc 64500ctgtccttcg cctatttccg ggtcaaggca tgcagcgacc tgtccgtcga gcaactgacc 64500

ttcggcgacc ggctccgcgt acggtcgaag gtgctgtcct gcggcggtga ctcgttgctc 64560ttcggcgacc ggctccgcgt acggtcgaag gtgctgtcct gcggcggtga ctcgttgctc 64560

accgtgcacc aggtcagccg ctatcacggg gacgccggta cggaggccgc gtacggcgac 64620accgtgcacc aggtcagccg ctatcacggg gacgccggta cggaggccgc gtacggcgac 64620

ggcctcgcca tggacgactt cttccgctac gactcgcccg gatgtgtgca cgtcgagacg 64680ggcctcgcca tggacgactt cttccgctac gactcgcccg gatgtgtgca cgtcgagacg 64680

ttcaaccgct ggatccagcg ttccggcgac ggcacgaacc acggattgcg gcgggcgacg 64740ttcaaccgct ggatccagcg ttccggcgac ggcacgaacc acggattgcg gcgggcgacg 64740

cccatcggct tccagtcgca ccacctggac ccgatcgccg acgagcacac cccccgtcgc 64800cccatcggct tccagtcgca ccacctggac ccgatcgccg acgagcacac cccccgtcgc 64800

gtcgtgtcga tggcgcgccg ggcagcgggc ttccgggccg agggggagcc gccgccggag 64860gtcgtgtcga tggcgcgccg ggcagcgggc ttccgggccg agggggagcc gccgccggag 64860

gccgggctca cgctgaccta ccaggtgagc gccagtcgcg acctcaacgg cgtcggcctg 64920gccgggctca cgctgaccta ccaggtgagc gccagtcgcg acctcaacgg cgtcggcctg 64920

ctgtacttcg cgtcgtactt ctcgatcgtc gactgggcgg tcctgacgtt gtggcgcagt 64980ctgtacttcg cgtcgtactt ctcgatcgtc gactgggcgg tcctgacgtt gtggcgcagt 64980

ctcggccggt cgtccgacag cttcctgcgc cggcgggtgc cggaccggca ggtctgcctg 65040ctcggccggt cgtccgacag cttcctgcgc cggcgggtgc cggaccggca ggtctgcctg 65040

ttcgccaacg cgaacgccga cgacgtcctc gacgtcgcgg tcacgacatt ccccgacgcc 65100ttcgccaacg cgaacgccga cgacgtcctc gacgtcgcgg tcacgacatt ccccgacgcc 65100

gggggtgagg acgtggtcga cgtgacggtg cgccggcagg acggcagcct gctcgccgtg 65160gggggtgagg acgtggtcga cgtgacggtg cgccggcagg acggcagcct gctcgccgtg 65160

gcgcgtcagc gcatcaccca gaaccgggcg ggatgagcac ggcgccccgc caccgggtgg 65220gcgcgtcagc gcatcaccca gaaccgggcg ggatgagcac ggcgccccgc caccgggtgg 65220

ggcgtcacgc cggctccagg cggctgttga tgaactcgac gagttcccgg ggggtccgga 65280ggcgtcacgc cggctccagg cggctgttga tgaactcgac gagttcccgg ggggtccgga 65280

cgaggcccag gtcgtcctcg gagatcggca caccgtagtc acgcttgatg cggctgtgcg 65340cgaggcccag gtcgtcctcg gagatcggca caccgtagtc acgcttgatg cggctgtgcg 65340

tctcgagcag ggcgagcgag tcgtacccca gctcgtcgaa cggttggtcc ggggcgtgct 65400tctcgagcag ggcgagcgag tcgtacccca gctcgtcgaa cggttggtcc ggggcgtgct 65400

cgaaggaccg ccagtccccg tcccagacgt actggcgcat gaggttctcg agctcccgca 65460cgaaggaccg ccagtccccg tcccagacgt actggcgcat gaggttctcg agctcccgca 65460

acgtgatcgg ctgcacgact ggctccttgt cctcgcagat cggggcgccg tccgccgcgc 65520acgtgatcgg ctgcacgact ggctccttgt cctcgcagat cggggcgccg tccgccgcgc 65520

ggccccgcca cacgcgctcg gcgcggcgtc gcaccgccgg acgtctggtc gagtgtgccg 65580ggccccgcca cacgcgctcg gcgcggcgtc gcaccgccgg acgtctggtc gagtgtgccg 65580

ccgtcccccg gagggccgct caaatgtgaa tggaacacga aagggggaat cgcgagagcg 65640ccgtcccccg gagggccgct caaatgtgaa tggaacacga aagggggaat cgcgagagcg 65640

tcggctggac gggccgatcg agtttcggca cgataaccgc gtgctcccgc agcaacagac 65700tcggctggac gggccgatcg agtttcggca cgataaccgc gtgctcccgc agcaacagac 65700

gcccgaggag gccgccgcca cggcgcgcct gcgccgtctc gcgcggctgc gccgggtgcg 65760gcccgaggag gccgccgcca cggcgcgcct gcgccgtctc gcgcggctgc gccgggtgcg 65760

tgaccggatc gaccgggagt acgcgcagcc cctcgacgtg caggcgctgg cgcgcggggt 65820tgaccggatc gaccgggagt acgcgcagcc cctcgacgtg caggcgctgg cgcgcggggt 65820

ggccatgtcg gccggttatc tcagccgcga gttccggctg gcgtacgggg agtcgccgta 65880ggccatgtcg gccggttatc tcagccgcga gttccggctg gcgtacgggg agtcgccgta 65880

cgcgtacctg ctgacccggc gggtggaacg cgcgatgacc ctgctgcgcc gcggtgacat 65940cgcgtacctg ctgacccggc gggtggaacg cgcgatgacc ctgctgcgcc gcggtgacat 65940

gaccgtcacc gaggtctgct tcgcggtcgg gtgtgcttct ctgggcacgt tcagcagccg 66000gaccgtcacc gaggtctgct tcgcggtcgg gtgtgcttct ctgggcacgt tcagcagccg 66000

gttcaccgag ctggtcgggg tgccgcccag tgcgtaccgg cgtgcggcgt cgcaggcgac 66060gttcaccgag ctggtcgggg tgccgcccag tgcgtaccgg cgtgcggcgt cgcaggcgac 66060

ggccgggatg ccgtcgtgcg tgtcgaagca ggtcacccgg ccgatccgga cccgggagga 66120ggccgggatg ccgtcgtgcg tgtcgaagca ggtcacccgg ccgatccgga cccgggagga 66120

gcggcaaccg gcctgacgac gcgagcttgc cttgagcgct gcgtgagcgt tgcggacgaa 66180gcggcaaccg gcctgacgac gcgagcttgc cttgagcgct gcgtgagcgt tgcggacgaa 66180

acttctgccg catcgccaac agaggagcct catcgtggtt ctcgccgccg gcccgacccg 66240acttctgccg catcgccaac agaggagcct catcgtggtt ctcgccgccg gcccgacccg 66240

gatccaggac agtgaccgcc gggatccggt ggtccgtctc cgccagctgt tcgacccggg 66300gatccaggac agtgaccgcc gggatccggt ggtccgtctc cgccagctgt tcgacccggg 66300

caccctccgg ctcgtccacg gggacgacga caccggcgtc gcggtcgtcc gcggccgcgt 66360caccctccgg ctcgtccacg gggacgacga caccggcgtc gcggtcgtcc gcggccgcgt 66360

ggccggcgga ccggtgatcg cgtactgcac cgacgcccgg atcatgggcg gcgccctggg 66420ggccggcgga ccggtgatcg cgtactgcac cgacgcccgg atcatgggcg gcgccctggg 66420

cgcggacggc tgccgccgca tcgtggccgc catccaggcg gcggtggtcg agcactgtcc 66480cgcggacggc tgccgccgca tcgtggccgc catccaggcg gcggtggtcg agcactgtcc 66480

ggtcgtcggg gtctggcact ccggcggcgc ccgtctgtcc gagggcgtgg cggcactcga 66540ggtcgtcggg gtctggcact ccggcggcgc ccgtctgtcc gagggcgtgg cggcactcga 66540

cggcgtcggc cggatgttcg ccgcgatggt gcacgcgtcc ggccgcgttc cgcagatctc 66600cggcgtcggc cggatgttcg ccgcgatggt gcacgcgtcc ggccgcgttc cgcagatctc 66600

cgtcgtgctc ggccccgccg ccggtggtgc ggcctacggc ccggcgctga ccgacgtggt 66660cgtcgtgctc ggccccgccg ccggtggtgc ggcctacggc ccggcgctga ccgacgtggt 66660

ggtgatgtcg tccgaggcgc gggtgttcgt caccggtccc gaggtggtcc gccgcgtcac 66720ggtgatgtcg tccgaggcgc gggtgttcgt caccggtccc gaggtggtcc gccgcgtcac 66720

cggggagcag gtcgacatgg agagcctcgg cggtccggac acccacggcc gccgctccgg 66780cggggagcag gtcgacatgg agagcctcgg cggtccggac acccacggcc gccgctccgg 66780

cgtcgcccat gtggtcgtcc ccaccgagag cgacgcgttc gcgcgggcgc gtgccctgac 66840cgtcgcccat gtggtcgtcc ccaccgagag cgacgcgttc gcgcgggcgc gtgccctgac 66840

gacgctgctc gggcagcagg ggacgtacga cgtggctgcc gtgcggacgc cgtcgaacct 66900gacgctgctc gggcagcagg ggacgtacga cgtggctgcc gtgcggacgc cgtcgaacct 66900

gtcggcgctg ctgccggaga accccaatcg cgcctacgac gtgcgtccac tggtccgggc 66960gtcggcgctg ctgccggaga accccaatcg cgcctacgac gtgcgtccac tggtccgggc 66960

gatcctcgat ccggcggagg agcacggttt cctggagttg cagccccgct gggcgccgaa 67020gatcctcgat ccggcggagg agcacggttt cctggagttg cagccccgct gggcgccgaa 67020

cgtcgtggtc gggctgggac gcctggccgg tcgcacgatc ggggtcgtgg ccaacaaccc 67080cgtcgtggtc gggctgggac gcctggccgg tcgcacgatc ggggtcgtgg ccaacaaccc 67080

gctgcgcaag ggcggatgcc tggatgcgct cggcgcggag aaggcggcca tgttcgtccg 67140gctgcgcaag ggcggatgcc tggatgcgct cggcgcggag aaggcggcca tgttcgtccg 67140

gcggtgcgac tccttcggcg tgccccttct cgtgctcgtc gacgtgcccg gttacctgcc 67200gcggtgcgac tccttcggcg tgccccttct cgtgctcgtc gacgtgcccg gttacctgcc 67200

cggcctcgac caggagtgga ccggtgtggt gcgccgtggc gcgaagctgt tgcacgcgtt 67260cggcctcgac caggagtgga ccggtgtggt gcgccgtggc gcgaagctgt tgcacgcgtt 67260

cgccgaggcg gtggtgcccc gggtcaccct cgtgacccgg aagtcctacg gcggcgcgta 67320cgccgaggcg gtggtgcccc gggtcaccct cgtgacccgg aagtcctacg gcggcgcgta 67320

catcgcgatg aactcccgcg cgctgggtgc gagcgcggtc ctggcctggc cgcaggcgga 67380catcgcgatg aactcccgcg cgctgggtgc gagcgcggtc ctggcctggc cgcaggcgga 67380

actggcggtg atgggcgcgg aggccgcggt ggccgtcctg caccgccgca ccctggccgc 67440actggcggtg atgggcgcgg aggccgcggt ggccgtcctg caccgccgca ccctggccgc 67440

cgcacccgac gacgagcgcg aggccctgcg gacgcagctg atcaaggagt acgagcagga 67500cgcacccgac gacgagcgcg aggccctgcg gacgcagctg atcaaggagt acgagcagga 67500

ctcgggcggg gtgcgacgcg ccctgtccat cggtgtcgtc gacgaggtca tcgatccctc 67560ctcgggcggg gtgcgacgcg ccctgtccat cggtgtcgtc gacgaggtca tcgatccctc 67560

gctcacccgg tcgaaggtgg tgacggcgct gatcgcggca ccggcccgcc ggggagcgca 67620gctcacccgg tcgaaggtgg tgacggcgct gatcgcggca ccggcccgcc ggggagcgca 67620

caagaacatc ccgctgtgac cgcgcggacc gtgacgccca cgtacggacc gacccacagc 67680caagaacatc ccgctgtgac cgcgcggacc gtgacgccca cgtacggacc gacccacagc 67680

cacagcttgc gaggaggact cgagtgaagg gcatcatcct ggccggcggt tcgggaacgc 67740cacagcttgc gaggaggact cgagtgaagg gcatcatcct ggccggcggt tcgggaacgc 67740

gcctgcaccc ggtgacgctg gcgatgtcga agcagctcct acccgtctac aacaagccga 67800gcctgcaccc ggtgacgctg gcgatgtcga agcagctcct acccgtctac aacaagccga 67800

tgatctacta cccgctctcg gtgctcatgc tcgccgacat ccgggacatc ctgatcatct 67860tgatctacta cccgctctcg gtgctcatgc tcgccgacat ccgggacatc ctgatcatct 67860

ccaccccgcg tgacctgccg ctcttcgaac ggctcctcgg cgacgggtcg cagttcgggc 67920ccaccccgcg tgacctgccg ctcttcgaac ggctcctcgg cgacgggtcg cagttcgggc 67920

tgtcgttgac ctacgctccc cagccggctc cccggggtct ggccgacgcc ttcatcatcg 67980tgtcgttgac ctacgctccc cagccggctc cccggggtct ggccgacgcc ttcatcatcg 67980

gcgcggagca cgtcggcgac gatccggtcg ccctgatcct cggcgacaac atcttccacg 68040gcgcggagca cgtcggcgac gatccggtcg ccctgatcct cggcgacaac atcttccacg 68040

gctacggctt ctccgaggtg ctgcaggcgg agagccgcga catcgacggc tgcgtcctct 68100gctacggctt ctccgaggtg ctgcaggcgg agagccgcga catcgacggc tgcgtcctct 68100

tcggctatcc ggtgaccgac cccgagcggt acggcgtggg cgagaccgac gcgacgggcc 68160tcggctatcc ggtgaccgac cccgagcggt acggcgtggg cgagaccgac gcgacgggcc 68160

ggctcatctc catcgaggaa aagccggaga agccgcggtc caaccgcgcc atcaccgggt 68220ggctcatctc catcgaggaa aagccggaga agccgcggtc caaccgcgcc atcaccgggt 68220

tgtacttcta cgacaacgac gtggtggaca tcgccaagaa cgtccgcccg tccgcgcgcg 68280tgtacttcta cgacaacgac gtggtggaca tcgccaagaa cgtccgcccg tccgcgcgcg 68280

gcgaaatcga gatcaccagg gtcaaccagg tctacctgga gcgcggcaag gcccggctgg 68340gcgaaatcga gatcaccagg gtcaaccagg tctacctgga gcgcggcaag gcccggctgg 68340

tcgacctcgg gcggggcctg gcctggttgg acgccggcac ctacgactcc ctgctgcagg 68400tcgacctcgg gcggggcctg gcctggttgg acgccggcac ctacgactcc ctgctgcagg 68400

ccgggcacta cctgcagacc ctggagcagc ggcaggggat ccacatcgcc tgcctggagg 68460ccgggcacta cctgcagacc ctggagcagc ggcaggggat ccacatcgcc tgcctggagg 68460

aggtcgccct gcggatgggc ttcatcgacg ccgacgcctg cctcagcctg ggcgcccagc 68520aggtcgccct gcggatgggc ttcatcgacg ccgacgcctg cctcagcctg ggcgcccagc 68520

tggcccacac cgagtacggc aggtacgtca tgaacgtggc ggcgggagcc cgctgacggg 68580tggcccacac cgagtacggc aggtacgtca tgaacgtggc ggcgggagcc cgctgacggg 68580

agtgccgaac ggcccggccg gtgcgatccg gccgggccgt ccggcgggtg acgccgtcac 68640agtgccgaac ggcccggccg gtgcgatccg gccgggccgt ccggcgggtg acgccgtcac 68640

accacctcga actgcgccat ctggccgagc tgcgagtgcc gcaggtagtg gcagtggtag 68700accacctcga actgcgccat ctggccgagc tgcgagtgcc gcaggtagtg gcagtggtag 68700

acgtagcgcc ccttgaaccc ggtgaacgtg gcctggatgc gcacggtgga accggggggc 68760acgtagcgcc ccttgaaccc ggtgaacgtg gcctggatgc gcacggtgga accggggggc 68760

accgcgacgg tgtccttcag gcccgcctcg cccggcgccg gcgggacgcc gtcgcggtcc 68820accgcgacgg tgtccttcag gcccgcctcg cccggcgccg gcgggacgcc gtcgcggtcc 68820

agcacccgga actgtacgag gtgcagatgc aggctgtggt cgatcgggaa gtccacgtcc 68880agcacccgga actgtacgag gtgcagatgc aggctgtggt cgatcgggaa gtccacgtcc 68880

acgttggtga cccgccagat ctcggtcgtg ccgatcggga tgcgggcgtc gacgcgggcg 68940acgttggtga cccgccagat ctcggtcgtg ccgatcggga tgcgggcgtc gacgcgggcg 68940

gcgtcgaaga cctcgccgtt gatggtgaac gtcgcctggc cgggccggcg gctcagcgtc 69000gcgtcgaaga cctcgccgtt gatggtgaac gtcgcctggc cgggccggcg gctcagcgtc 69000

acgtcgcgga ccgccgtggc ccggggcagc ggcggcagcg gccggagctt cgccgggagc 69060acgtcgcgga ccgccgtggc ccggggcagc ggcggcagcg gccggagctt cgccgggagc 69060

cggctgttgt cggcggccgc gccgaccacg tcgaacgcga gcaccgggcc ggtggtgtcc 69120cggctgttgt cggcggccgc gccgaccacg tcgaacgcga gcaccgggcc ggtggtgtcc 69120

tccaggacca ggcggctgcc gaccggtgac cggcggaagt ccacgacgat ctcggcgcgt 69180tccaggacca ggcggctgcc gaccggtgac cggcggaagt ccacgacgat ctcggcgcgt 69180

tcgcccggcg acagggccag ttcggtacgg ggcaccggcg cgggaagcag gccgccgtcg 69240tcgcccggcg acagggccag ttcggtacgg ggcaccggcg cgggaagcag gccgccgtcg 69240

gtggcgacct ggatcatgtc ggcgccggtc aggctcagct ggaagttccg ctggatggcg 69300gtggcgacct ggatcatgtc ggcgccggtc aggctcagct ggaagttccg ctggatggcg 69300

ccgttgagca gccgcaggcg gtaccggcgg gcggccaccc gcaggaccgg ctgtggtttg 69360ccgttgagca gccgcaggcg gtaccggcgg gcggccaccc gcaggaccgg ctgtggtttg 69360

ccgttcgcca gggtggtctg ctggacgaag ggatccgccg ggtcgaacgc cagcgtgccg 69420ccgttcgcca gggtggtctg ctggacgaag ggatccgccg ggtcgaacgc cagcgtgccg 69420

tcggccgcga tccgggcgtc ccgcagcatg atcgggatgt cgtaccggcc cgtcggcaac 69480tcggccgcga tccgggcgtc ccgcagcatg atcgggatgt cgtaccggcc cgtcggcaac 69480

ccgaatcgcc gttcggcgtc gtcgtcgatc aggtagaacc cgtgcaggcc gcggtagacg 69540ccgaatcgcc gttcggcgtc gtcgtcgatc aggtagaacc cgtgcaggcc gcggtagacg 69540

tgctcggctt ccatgtggtg gctgtggtcg tggtaccaca gcgtcgcgcc ctgctgccgg 69600tgctcggctt ccatgtggtg gctgtggtcg tggtaccaca gcgtcgcgcc ctgctgccgg 69600

ttcggatagt ggtaggtgcg caggcgcccc ggctcgacca cgtccatcgg gtgcccgtcg 69660ttcggatagt ggtaggtgcg caggcgcccc ggctcgacca cgtccatcgg gtgcccgtcg 69660

ctggcggcgg cgaccgatgc gccgtgcagg tggacgttcg tcgggtggtg caggccgttg 69720ctggcggcgg cgaccgatgc gccgtgcagg tggacgttcg tcgggtggtg caggccgttg 69720

ttcagccgga cgatcgccgg ccggccgcgc cgggcgcgga tggtcgggcc gacgaaccgg 69780ttcagccgga cgatcgccgg ccggccgcgc cgggcgcgga tggtcgggcc gacgaaccgg 69780

ccgttgtagc cgagcaccgg tgtcctcacc ccgggcagga tctccgtcgt ggtggaccgg 69840ccgttgtagc cgagcaccgg tgtcctcacc ccgggcagga tctccgtcgt ggtggaccgg 69840

accgacaggt cgtagacgtc ggtgccgccc accaccgcgc tcgggcgcag taccggagga 69900accgacaggt cgtagacgtc ggtgccgccc accaccgcgc tcgggcgcag taccggagga 69900

accggaagcg ggacggtgaa ggggacgggc ggggtgctgg ggtcgccggt gccgccgccc 69960accggaagcg ggacggtgaa ggggacgggc ggggtgctgg ggtcgccggt gccgccgccc 69960

gggtgcgggc cgtgtccggc atggatcgcg gcgctgctct 70000gggtgcgggc cgtgtccggc atggatcgcg gcgctgctct 70000

Claims (6)

1. An anthraquinone oxidase gene nes27, which is characterized in that the nucleotide sequence is shown as the base sequence of 32561-33304 of SEQ ID NO. 1.

2. An anthraquinone oxidase NES27 encoded by the anthraquinone oxidase gene NES27 of claim 1.

3. The use of the anthraquinone oxidase NES27 as claimed in claim 2 in catalyzing the formation of the compound ethyl-kinebscriptione of formula 2 from the compound ethyl-dehydrorabemomycin of formula 1

4. A genetically engineered bacterium lacking the anthraquinone oxidase gene nes27 of claim 1.

5. The genetically engineered bacterium of claim 4, wherein the genetically engineered bacterium is Micromonospora echinospora (Micromonospora echinospora).

6. Application of genetically engineered bacterium of claim 4 in preparation of compound ethyl-dehydrorabemomycin shown in formula 1

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