CN110373477B - 克隆自cnv片段的与猪耳形性状相关的分子标记 - Google Patents

Abstract

本发明属于猪分子标记筛选技术领域，具体涉及克隆自CNV片段的与猪耳形性状相关的分子标记。该分子标记为通过整合NCBI数据库和欧洲生物信息研究所公开的包含58个品种的469头猪的重测序数据，对比猪参考基因组序列获得基因分型数据，根据全基因组关联分析(GWAS)技术得到一种与猪耳形性状关联的拷贝数变异分子标记(CNV)，当个体拥有该片段的多拷贝(拷贝数>3)时，则有利于猪拥有非直立的耳形，该片段的核苷酸序列如序列表中所示。公开了筛选与猪耳形相关分子标记的方法及其关联分析方法的应用。本发明为猪耳形的标记辅助选择提供了遗传标记资源。

Description

克隆自CNV片段的与猪耳形性状相关的分子标记

技术领域

本发明属于猪分子标记筛选技术领域，具体涉及克隆自CNV片段的与猪耳形性状相关的分子标记及其应用。

背景技术

猪是家畜生产中重要的经济动物。家猪作为人类最早驯化的家养动物之一，起源于欧亚野猪(Sus scrofa)，随后欧亚野猪在欧洲和亚洲分别被独立驯化(Larson G,DobneyK,Albarella U,et al.Worldwide phylogeography of wild boar reveals multiplecenters of pig domestication[J].Science,2005,307(5715):1618-1621.)。由于这种独立驯化过程，欧洲猪种和亚洲猪种之间有着区别分明的种质特色，例如欧洲猪种拥有更高的瘦肉率和更大的体型，亚洲猪种则脂肪含量更高，并且拥有高产仔数和早熟的优点。

耳朵形状是猪重要的外表构象，是一个品种的独特商标(Wei W H,De Koning DJ,Penman J C,et al.QTL modulating ear size and erectness in pigs[J].Animalgenetics,2007,38(3):222-226.),目前为止，关于耳朵形状的相关研究较少，因此，探索发现耳形遗传机制的相关分子标记有助于更好的展开猪的育种研究。

全基因组关联分析(Genome-wide association study,GWAS)作为一种关联分析方法，在复杂性状的相关研究中具有比连锁分析效果更好，统计效力更高的优点，最初是1996年由Risch首次提出的(Risch N,Merikangas K.The future of genetic studies ofcomplex human diseases.[J].Science,1996,273(3):350-354.)作为在全基因组范围内探索鉴定实验群体中和目标表型相关联的遗传标记的有力工具，全基因组关联分析已经被广泛应用于各类研究，其中包括人类疾病以及家畜重要经济性状等等(Visscher P M,Brown M A,McCarthy M I,et al.Five years of GWAS discovery[J].The AmericanJournal of Human Genetics,2012,90(1):7-24.)

拷贝数变异(copy number variation，CNV)是一种片段长度在1kb甚至1Mb的遗传标记，对比于参考基因组上的片段，该部分可能发生不同程度的缺失或拷贝数的增加。而由于该片段拷贝数的缺失或增加，可能会引起显著的基因表达量的变化，从而影响到基因控制的对应表型(Zhang F,Gu W,Hurles M E,et al.Copy number variation in humanhealth,disease,and evolution[J].Annual review of genomics and human genetics,2009,10:451-481.)。

迄今为止，对于猪耳形性状相关的分子标记及其应用的报道较少。

发明内容

本发明的目的在于寻找与猪与耳形性状相关的分子标记，通过将猪重测序数据比对到猪参考基因组上，获得SNP分型数据与CNV数据，并使用全基因组关联分析(GWAS)筛选与猪耳形相关的CNV片段，为猪耳形的相关育种提供新的分子标记资源。

本发明通过整合NCBI数据库(SRA,http://www.ncbi.nlm.nih.gov/sra/)和欧洲生物信息研究所(EMBL-EBI,https://www.ebi.ac.uk/)中公开的包含58个品种的469头猪的重测序数据，使用MVP软件，利用MLM模型(Zhang Z,Ersoz E,Lai C Q,et al.Mixedlinear model approach adapted for genome-wide association studies[J].Naturegenetics,2010,42(4):355.)，筛选出与耳形相关的CNV片段，为猪耳形的辅助选择和耳形预测改良提供了新的分子标记基础，对于猪的育种具有重要意义。

本发明筛选的CNV片段与猪耳形的分子标记相关性达到了显著水平，为猪的耳形相关研究提供了新的遗传资源。

本发明的技术方案如下所述：

申请人通过整合NCBI数据库(SRA,http://www.ncbi.nlm.nih.gov/sra/)和欧洲生物信息研究所(EMBL-EBI,https://www.ebi.ac.uk/)中公开的包含58个品种的469头猪的重测序数据，并将猪重测序数据(核苷酸序列见序列表SEQ ID NO:1)比对到猪参考基因组(基因组版本11.1,Sscrofa11.1)，当个体拥有该片段的多拷贝(拷贝数>3)时，有利于猪拥有非直立的耳形。

上述片段可以作为检测与猪耳形性状相关的分子标记。

申请人提供了一种筛选猪耳形性状相关的CNV分子标记的方法，所述的方法包括如下步骤：

1.整合了NCBI数据库(SRA,http://www.ncbi.nlm.nih.gov/sra/)和欧洲生物信息研究所(EMBL-EBI,https://www.ebi.ac.uk/)中公开的包含58个品种的469头猪的重测序数据，并将猪重测序数据比对到猪参考基因组(基因组版本11.1，Sscrofa11.1)上，获得SNP分型数据；另将重测序数据进行初步质控后，使用相关生物信息软件获得CNV数据。

2.根据群体中个体对应的品种信息，通过查阅中国畜禽遗传资源志(猪志)，维基百科，国外相关猪品种网站，猪遗传资源相关文献等，统计了每个品种的特色耳形，并将耳形作为表型；

3.采用混合线性模型的方法，利用R统计环境下MVP软件包中的MLM模型进行GWAS分析。具体模型如下：y＝Xβ+Zμ+e，其中，y代表个体的表型值，β代表包括主成分(principlecomponent)在内的固定效应，μ代表服从分布u～N

的随机效应，该分布中G代表由对应个体的SNP计算的亲缘关系矩阵。X和Z代表β和μ的关联矩阵,e代表残差向量(vector ofresidual errors)。

本发明筛选的分子标记可应用于非诊断目的的对猪耳形相关基因的基因型或猪耳形性状相关的关联分析中，为猪的耳形性状的分子标记辅助选择提供了新的分子标记资源。

本发明与现有技术相比具有的有益效果：

本发明可通过在体外采用基因芯片技术检测猪的基因型，作为非诊断目的的评价猪的耳形，与目前的PCR-RFLP等方法相比，本发明具有简单、快捷、灵敏度高和特异性好等突出优点。

更详细的技术方案请参见《具体实施方式》。

附图说明

图1：本发明的总体技术流程示意图。

图2：是本发明的曼哈顿图。附图标记说明：黑色圆圈及箭头指向标记的为本发明筛选的分子标记，该标记位于猪第5号染色体上。

具体实施方式

对序列表的说明：

序列表SEQ ID NO:1的是本发明检测到的与猪耳形性状相关的的CNV分子标记的核苷酸序列。

本发明中的序列和全基因组关联分析结果，是基于猪基因组11.1版本。

实施例1：基因分型检测

(1)整合下载NCBI数据库(SRA,http://www.ncbi.nlm.nih.gov/sra/)和欧洲生物信息研究所(EMBL-EBI,https://www.ebi.ac.uk/)中公开的包含58个品种的469头猪的重测序数据。

(2)使用SRAToolkit(V2.8.2)将原始数据转换为fastq文件，随后使用Trimmomatic(V0.36)软件对fastq文件进行初步质控。该步骤后通过使用Burrows–WheelerAligner 0.7.17(BWA)软件将剩余的高质量reads与猪参考基因组(版本11.1)进行比对，取唯一比对序列使用GATK分型。为了获得高质量的变异数据，使用GATK(V4.0.3.0)软件通过‘QUAL<30.0||QD<2.0||FS>60.0||MQ<40.0||SOR>4.0||ReadPosRankSum<-8.0’命令进行质控，获得可用于GWAS分析的基因型数据。

(3)去除掉重测序数据缺失率在5％以上的个体，随后使用CNV caller软件对重测序数据进行分析，以获得该群体的CNVR数据，经过质控和校正后，保留了全基因组变异百分比在30％以内的321头个体，并获得了18016个CNV用于后续分析。

(4)根据对应表型个体，最终有319头个体，18016个CNV用于GWAS分析。

实施例2：Chr 5:29826401-29866000片段在猪耳形性状关联分析中的应用

Chr 5:29826401-29866000片段与耳形性状关联分析：

用于基因型与耳形性状关联分析的表型主要来源根据群体中个体对应的品种信息，通过查阅中国畜禽遗传资源志(猪志)，维基百科，国外相关猪品种网站，猪遗传资源相关文献等，统计了每个品种的特色耳形，并将耳形作为表型，有CNV基因型的共计319个个体。品种的特异耳形根据统计结果共分为两个类别，即直立(如大白猪，汉普夏猪等)和非直立(如长白猪，梅山猪等)，其中直立的个体表型编号为1，非直立的个体表型编号为0。采用混合线性模型的方法，利用R统计环境下MVP软件包中的MLM模型进行GWAS分析。具体模型如下：y＝Xβ+Zμ+e，其中，y代表个体的表型值，β代表包括主成分(principle component)在内的固定效应，μ代表服从分布u～N

的随机效应，该分布中G代表由对应个体的SNP计算的亲缘关系矩阵。X和Z代表β和μ的关联矩阵,e代表残差向量(vector of residualerrors)。该标记基因型与对应表型个体数见表1，全基因组显著水平见表2。

表1 Chr 5:29826401-29866000片段的拷贝数及对应个体数

表2 Chr 5:29826401-29866000片段的全基因组显著水平

表2说明：当标记的P值P<0.05/18016(Bonferroni校正)时为显著标记

由表1可知，对于Chr 5:29826401-29866000片段拥有多拷贝(当拷贝数大于3时)的个体，有利于猪拥有非直立的耳形。

主要参考文献：

[1]Larson G,Dobney K,Albarella U,et al.Worldwide phylogeography ofwild boar reveals multiple centers of pig domestication[J].Science,2005,307(5715):1618-1621.

[2]Wei W H,De Koning D J,Penman J C,et al.QTL modulating ear size anderectness in pigs[J].Animal genetics,2007,38(3):222-226.

[3]Risch N,Merikangas K.The future of genetic studies of complexhuman diseases.[J].Science,1996,273(3):350-354.

[4]Visscher P M,Brown M A,McCarthy M I,et al.Five years of GWASdiscovery[J].The American Journal of Human Genetics,2012,90(1):7-24.

[5]Zhang F,Gu W,Hurles M E,et al.Copy number variation in humanhealth,disease,and evolution[J].Annual review of genomics and human genetics,2009,10:451-481.

[6]Zhang Z,Ersoz E,Lai C Q,et al.Mixed linear model approach adaptedfor genome-wide association studies[J].Nature genetics,2010,42(4):355。

序列表

<110> 华中农业大学

<120> 克隆自CNV片段的与猪耳形性状相关的分子标记

<141> 2019-07-11

<160> 1

<170> SIPOSequenceListing 1.0

<210> 1

<211> 39600

<212> DNA

<213> 猪(Sus scrofa)

<220>

<221> gene

<222> (1)..(39600)

<400> 1

cttttttagt agtagttatt tttaaaagtg ctatacaaga attttcttca gaaccatcat 60

aactaaaaat taaagaggag aaagttagac tttgaggtac aatggcaaga tgaaaagaca 120

tgttccaccc atatgctagc aaattccaat caaaataacc aaagcaatgt aaaataattt 180

ttttttcctt tgtctttttg catttttgcc ttttctaggg ctgctctcac agcatatgga 240

ggttcccagg ctaggtttcc aatcggagct gtagctgccg gcctgcgcca cagccacagc 300

aatgcaggat cccagccacg tctgcaacct acaccatagc tcatggcaag gccggatcct 360

taatccgcag agcaaaaaca gggatcgaac cctcaacctc atggttacta gttggatttg 420

tttccgctgc gccacagtgg gaactcccag aacttgcatt ttggaagtca tttcaccact 480

catcctcgaa caagactgag aagccttgaa tggagaggtc taaaggcaat tatcatccca 540

caacataagt aaggatttgg actccttaaa gggccaggca gcagctagcc accagacaga 600

cactgtggaa agtaacattt ctatcaaagg aaatcttggt ccagtttcag ctgaaacgcc 660

ttcctgcgac ccccctcctc tttccctctt ctcccggtaa taatttgttt caaattatcc 720

aactgagaag agtatgtgcc ctgacctgac caatgggaag gggacaggta catcgcttgc 780

actagggata aatagggagg gtcttttctt ctttgcgcac actttttgga gtactcgcgc 840

ccttctgcag aagtaaagag ccttgtcgag atctccccgg gttcacctgt ctcatttttc 900

gatactgatg atccgagcca cgtccccaac aacaccagac atggtcccat agtatactgg 960

gaaagaaggc caggccaaaa agtaaattat gtgtggaaaa aaatgttcct tggcataaaa 1020

gatttgccaa acacagtgaa gggtatacac agacaaatgt ccccagtgag gacaacctgt 1080

aacatctgct tcctcctatt tccctcagtg tacctggatt attggactgg taatagaatt 1140

gttctccaag ccccagtcac ccacaccaga aatttttaag gtcaaatttc agtaattctg 1200

atgccagtct tttctttacc catcacatac tgagcgagac cctgggtgca cgcttctatc 1260

ccctcacaca cagcctcaga ggacatttct gaatgtcctc acacctctct gctttactaa 1320

tgcttctcaa aggttggtgt gtgatgtcga tagctttcct gactgaaaaa aatattactt 1380

actttttcaa aaacttattt aatttcattt gatttctgct gaaagaaatc ttggtgtgca 1440

tctttgctgc ttctttgacc agagtttgct ttttggcctt tgcttttctt tctctacaag 1500

gtattttcct ggatccatac cccacagccc caaaacatag gtccactcta tttgtattgc 1560

cctggtgcct agctagatgt ggcatctcag gctaatctgc agagagaaac tccatcattt 1620

ccctggattc tttccaaagg ctggcataag atctttagaa aatctttagt agtggagaat 1680

ctttgctctt tgaggtgctt aattttctaa aaatgcttaa attacccttg cagtggtctt 1740

tgtggaatct tctttggttt aaagcctatg ttttacagtg tttgggttct gctctttctc 1800

cttttttgaa aattgggcaa atggattctt attgtttact ttgtgtctat acctgtgagc 1860

tggtatccat gtcaacatct gactctacaa gcgcttgact ctatcctctg acttgcaccc 1920

tgcttctttg gagttgacat tcctgctgtg atctttctat cttaattctg atacctcctt 1980

tagaatttgg cttggatcta ctcttctcac tctgttctag tatattcttt agaacagtct 2040

ctgtgctgta tgcagatagc ttaaaatcta accgacaaat agcttagtat ctgcctaaca 2100

tagcaaaaca acagacagta atttaaaatg gccattgtat aggtttcagg ttgtataata 2160

gagttgttag gtgttataga cattcggaaa tagtggggaa tagttatgga gcacttcagg 2220

ggaacatata gaacttcaac aaggtatttg tgcatggctg ggatttggag atggcaaaat 2280

tccagtcaag tgaatgattg tgaatgaggg aagaatgtgg taatgagctt aggcatattg 2340

atgcaaccat aaaaatatac tcctcagcat actccatctt tccataaatg agatagcaaa 2400

taggggctac gggacaacaa atagtagagc tgtgaaagtg atttgggcag agagagcctt 2460

gtgtctggga ggctaattaa atattatcta gttggaaaat agtgaggctt caaattttga 2520

tagtggctga ggaatggaga aggagggatg gaaattagag atagtggtcg atacaggagt 2580

taatgactaa gtagatgtgg gagtataaca gatgagtttt gactattggt aaataaaaca 2640

atggcgatac catcattaga ggcaggaaaa tatacgtaag tcaaattgct gaatttgtta 2700

tttcaatcga gttacttttg ttttataagt taaggtaaag acctctttct gactccttgc 2760

acttttccca tttaaaattt tgattgttgt ttctttttca tctttcggcc aaacccaaat 2820

ctatatttaa aaatcttttt atgtatttca agactatttt tttttttttt tttgtctttt 2880

tgccatttct tgggccgctc ctgcggcatg tggaggttcc caggctaggg gtccaatcgg 2940

agctgtagcc accggcctac accagagcca cagcaacgcg ggatccgagc cacgtctgca 3000

acctacacca cagctcacgg caacgccaga tccttaaccc actgagcaag gccagggatc 3060

gaacccgcaa cctcatggtt cccagtcgga ttcattaacc actgtgccac gacgggaact 3120

ccttcaagac tattttgagt ttagctttcc atttgtcagg ctgcacaatc ctaggtactc 3180

acttctttat ccatttgtat gggccattaa aaaaaaaaca aaaacaaaca aaaaaacact 3240

ttccttatat ttgtgattat cttgtgaatc tttttcttta tccacataac tctggatgct 3300

aaattcagat ttggtcataa tacttcttta tagttacagc ctgaacaata ttgattttga 3360

ataaagaata gtacgtcaaa tttctgaaaa tctgtacacc tgtttattca tagcagtaaa 3420

gtgctatact acgctgataa gccaggttca agttgtcatg ttttctgcaa agcactttga 3480

aagattttgg taacattttg tgaaagtttc tgggaaagtg ttgatggact tctaggcaac 3540

attcttcctc ctctgaaaaa taaagggctt tgataagaaa gcagtttatc aaggtggaag 3600

taaaagataa gacagggcta gtaacccaag gtctatggac ttctgaaatt ttggtatatg 3660

catttggatt aatagttttt tggagagggt ccagatattt cagccatttt tcaataatcc 3720

tcaatgatct cgagaagtag attttttatt tcgaattaca gaaggagatg aggctaagtg 3780

aggttatagt atttgtcaga gatctcacag ataacaaatt gcacagctag gatttgaacc 3840

tagttgtatg tgtgcatgta tgtataaatt aaaattcaaa ttccatgctt ttccctctta 3900

gtcaaactga ttctattcta tcactttgct atacacctgg aactaacaca atattttaaa 3960

tcaactatac ttcagtataa aaaaatttct attctaggaa aagtcatgat aatcagcata 4020

gggcggattg agaagtttca aaacagtgtc catggaatac cgaggaatag gccttagata 4080

aggagttctt agcttgggcc ctttggattg attttaaagg gtctgtgtat ctggaaggga 4140

ggaaaaaaaa aaaaaccaac atattcactt tcactaaccc caatttctat taattataaa 4200

tgtagataat aacctaccat agtagttgtg actatatcac cagtagaaat cctatatttt 4260

cttatcatat tacaattgtc acagatatct cacagtaaca tttatactca taaatgcttc 4320

aaagttatgg caattactag agccaccatt agagcttgtt ttgtaatgtg ttgataaaaa 4380

aacagatgta ctactatact gtaaatttgg tggttttttt tttttttttt ggtattttga 4440

aaacactatt taaagttgtt tcttctctgt aatcccatgt atttcctctc attcatttaa 4500

aaacattatc ctgagtggtc tgtagtcttc accagacttc caaaaacctc atggaacaaa 4560

aaaagtttaa gaacccctgc tttgagtgtt atattgtcat tttatcattc ccaaggggtt 4620

tttcttgttt gttttttttg ttgttgttgt tgtttgtttt taaagtacat tgaacatggg 4680

atatgcagct caggatttca tacatataac ttattgaaaa aatatttatc aaaatcacta 4740

atcatcaggt agaatgtaga ctttttggct ttcaggaaat gtttgtagct tcttattatt 4800

ttatttcttt tcatcactag ttgtattctg tcatagacta tttcagcttg acaactctct 4860

gggctgcctc aaagacaggt tcatgtagtg tttaaaaata aggacttgga aacaaacaca 4920

ggttcacagg ccagcttcac cacttaagag tgcatggtaa aatgaccttg tggaagatat 4980

atatataatt tttcaacttc agtctctcca cctgtaaatg ggaataagta gggttgctgt 5040

agggtatata taagaaatgt aaaccactta gcaaagcatc tgacaagtaa tagcaataat 5100

attcattgaa gttggtctta aataagactt gctaccctta tattctcaat cccagcttct 5160

ctgaatctcc tgagctaaat agtgagggaa atcttgggta tatggagaat ttctcttatt 5220

cttatccttt cattccttca gcaagatctg gcaggcatct gcctctctgc ttccatcatt 5280

ggcaggacta ccagcttctg gattacttaa ataacaagtc ctaagcccaa gtgagccaat 5340

aaagaataac cataacacat gtagaggagt ccaacatgca gataggatca ataaagataa 5400

atagacacac atcttatgaa gtagaacctc tggaggagac caataacaac taaaccaaaa 5460

caacaacaat agaaaactgt gatgacgttg gctgtctaat acaattatat tcatcttttt 5520

ttttaattag ggcagggtga cttccctccc caggtgaata ataactaatc atgattggat 5580

tcctaatcat aacttatact aatcgtaaaa ttgaattcct cttgtcagca gctgatttca 5640

gccatgcagg tactccattc ctggtcagtt gcatctcagg ggaaatctga ttggaggttc 5700

ctggtaaaag tttccctcgt tgctaaaaag agacatacta gttgaagtgc ctccattttc 5760

actcatgctc tcagatctgc ctgtggtgtc tggaattgct gcagccattg tgtgaccatg 5820

agaggagatc actgacattg tgagggggta gaaaggaaaa cacattggcc tttatgagta 5880

gctgtgccac ttacttcacc aacatgggac cacctaaata tattttcaaa gccactttta 5940

ttttgtgtca tttctttcag ctgaaatatc ctatttgata caattaaagg actttatgtc 6000

cagtcccctc agaagtggag atttgaaaat atgatttccc agctaaacaa ttcagtaaaa 6060

agtgaaatga aggaagccac tgctgagacc caaattggag gagatatctg aaaggacgta 6120

gtgaaaaata aagaagtctg tatcatgaag gaaaaattat gagacttgga agatagctcc 6180

aggaggctta acatgtaaat aacagatatt tgaaaaaatt tctttttctg accaaaagat 6240

tcaagtctgc gtagaagaaa aatatctttt ttctctagta ttctaagttc taaccttacg 6300

ctcccatcac aaagattaac aagagcaaaa cagggaattc ccacggaaac aatctcaact 6360

agtatccatg aggatgcagg ttcaatccct gcccttgctt ggtgagttaa ggatcttgtg 6420

ttgccacaag ctctggtggc acacgcagct cagatctggc attgctgtgg ctgtggtgta 6480

ggccggcagc tgtaggtccg attccctagc ctgggaactt tcatatgcca cacctgtggc 6540

cctaagggaa aaaaaggggg tgtggggaga ggaacaaatt tatttaaaaa tatttttaaa 6600

atactttaca tgacacaaaa gcctttataa ggagacccag aaaagcagtt aagcttgaat 6660

gttttcatgc taggtttgag gaaaagcgga gagtcttggg aagatgtgag aagacaaaga 6720

ttatgagcca gcaaggcctg ttcattccaa ttcctttgat tcctttctga gccttggaga 6780

tgaagatatt cttttttctc taggtttagg gaaagcacct ctcatacgaa ggttttgtga 6840

cctgcttcag aggacattta tttctcagat tccatttccc aggttccttc agcttatgtt 6900

cagtgtgcca atgtgccata gttggggata gcacatccca tgctccatca cctgcagatt 6960

gaacaggctc accaaattgc gcataggatt aatgagaaac cctgggcacc caaagcctta 7020

cttgatgctt caaggataaa gaggaaattt tataggcttc caaatttata agtgtcagat 7080

tagccttaga tttctcaacc ctggactgga agatggcagg ataacctgta tagaagggga 7140

gggaaggcag agaggaggaa aatgagtttt ctaaaggttt aatctcacat gggaagaaag 7200

ggaagctata gcatcttgat ctgcaaaaca atgggtcttt ggtgaggaaa ttgtaccttg 7260

tatatagtat attgtttata gtagctaatt ctgtttgtga gtgcagagga agggagaatg 7320

acttaagggg gggaaatgga ataatagcaa atggcagaaa caagggggaa aacacaacaa 7380

aaaaagaatg gtaaatgtga agaaagatga aacaagagga gttcccgttg tggcacagtg 7440

gttaacgaat ccgactagga actatgaggt tgtgggttca atccctggcc ttgctcagtg 7500

ggttaaggat ctggccttgc catgagctgt ggtgtaggtt gcagatgaag ctcagatccc 7560

gcattgctgt ggctttggcg gctggcggct gcagctctga ttcgacccct agcctgggaa 7620

cctctatata ctgtgggagc agcccaagaa aaggcaaaaa gacaaaaaaa aaaaaaaaaa 7680

aaaaaaatta aattgtcaaa tcaaatatga atgagctgaa tttccccatt aaaaaaaaca 7740

gagattatca gagattgaat taaaaattta gctatttgct acttagaaaa ataaacaaaa 7800

tatcaaaaaa gcttaacaat aaaaagattg tcaaagggta ttaggtcaaa gaaaacaaaa 7860

gggaaagtag aaatgaccat attactatca aagtataatt taaactaaaa agcacaaaag 7920

gataaaaatg taaattatgt gatattaaaa gggcattatt tataaagagc atagagcagt 7980

tagaaacctg taggcataaa ataacgtaat gactaaatat atgaacaaga atcttagaaa 8040

tgaaagaaaa caattaaaaa atacaaatta taatggataa cttcactctc cttccgactg 8100

agatatgtga gtagtcataa ccaagtaaag actaaccgaa taatatagtc aataaacttt 8160

atttagtaga gatataaagt atcttttaaa tttaaaaaat ccctcaaata aaaaatgaaa 8220

ttttttattt tcatgggaca ttaaaaaata gatcatttac ttgattgtaa agaaaatatt 8280

aataaatttt taaaagcaga gatttctaga ggccatattt ttaagataat ccaatgaaat 8340

tagaaataaa tgtgggaata cagtgaacat tatacttagg ggtaaaatta caaccttgaa 8400

taccttcatt attaaaagaa aaaaagatga aaaagaaagg aattttatca aaagaacaat 8460

aaaaataata ggaaaatgga ataaagataa aagctgcaat taaaatgaaa tataagacat 8520

taaacaattg taggaagaat aaatgaattc aaagttgatt ctttgaaaga aagaaaaaat 8580

agattaactt atactgggca cattaagaga aaaaaagccc aacccataat atttatgaaa 8640

gagaaaacta taagtagata gttgcctttc ttggcatgag aacatgctat gaagtcacca 8700

caataaaatt atattttcat agaaatagat atgaagatca gtggagcatc tacttgagaa 8760

tctataaaga tcagttaaac atggtatttt aactcagtca ggaatagata atgtaaaaat 8820

tgttgacggc ccaattgctc ttccttttgg aagaaaataa aatcgaaata ttacctcaca 8880

tactgaaaga aattacagat ggattaagat ctaagtgcaa cccccccccc aaaaaaaaac 8940

aactcctgca tgaaatgtag gatgccatgt gtataatcca aaggcaggaa actcttctta 9000

atcaaggcca ggagcccagg aagtgcacag aaaagattag acatatttga ctgacaaaat 9060

cataagccaa gtccacacag aagcaacata tataggagaa taattgtaat gcaaaggcca 9120

gtaattaatg tctgcagtat acactgcagt atgcagtgtt accaattgtg aagaaaaaca 9180

atagtaacaa ataactctat agaaaaaaag ggtgaaaata tggagttaca aattccaaaa 9240

aaggaaatgc caacaaacat taagaactgt gtatcttgta tcttgggagt tcccgttgtg 9300

gctcagcagt aatgaacctg actagtgtcc atgaggatgt agattcgctc tctggcctca 9360

ctcagtgggt tggggatctg gcattgcctt gagctgtggt gtaggtcaca gatgaggctt 9420

ggatctggtg ttgctgtagc tgtggtgtag gctggcagct gtagctctgc ttcgacccct 9480

agcctgggaa ctttcatatg ccacaggtgc gtccctaaaa gaaaaaaaaa aaaaaaaaag 9540

aattgtctat cttatagtca atgaaacaca aatttttctt atatgaacta ctggcaataa 9600

tttaatggaa atttttaaaa gctgcaacac caattgttga tggagggtat ggaaaagaag 9660

tatttgacta tatttgttat ggaaagccat ctagtaatat cttctaaaca caaaaacaca 9720

tttatacttt gacctagtag tcctattctt aagaatcaat cccataggaa gaaaaattct 9780

accagatgta atagcatatg tataaggttg ttattgcagt atatttcatg gaggaaaaac 9840

aatcagaaac aaagtcattc ttgtcatttg tgggatggtt gcaaagtgga atattatgcg 9900

tctactagaa catttatgtt tatgtgtgtg tagctactta tgattttgta aggctataag 9960

aacatgggac aaaaatgtgg tagaaccctt attaggttgt tagcatagtt tccttgcagg 10020

atgaggaaag gggccaagaa aactggggga aaaaattttc atttgaaata cgtagggtat 10080

attcacattt gcgcatttgt gtatggttaa tagatgtgta tatgatatat ttaaaaagta 10140

acaggagcta agcaacaagg cattaggaga aaaataacat taagatatct tggaggccag 10200

aaacccaggg aaatatctgg aactgagcaa aaggttgaga ttgaggggta taaattattt 10260

tattataaat aaaaatgcat gaaaacaaaa actgttaatt atcaaagagc tactatactg 10320

gctctggatt tatacctgaa gtcaaagtag ataattttaa aaaatctatt atatagcacc 10380

agaccttggt gatattcccc aggggagttt cctaatatgg gaaatatagt taaaatttat 10440

ttttgcctag cctttaagac catgattttg gccataatat tacatggctg agacttaaat 10500

aggatgtggt cgaatgcttt atgtctatgg atgctgacca caccttctaa aatattatgc 10560

tccccaaatg ggaaaatatc aaagaaatta tgtaggtgcc catgatgtac tatcgagaga 10620

gtaatccatg gaatgctata ctctggtata gtgacataag aaataataaa tacttgagag 10680

gtgatcattg tcaaaaaatg cttgatatat atatatacgt cgcttaattg tggtatcatt 10740

ttcacaattt ttgttaatac tttcttcagc ttcaatttgg ttcacttatt caacaaatat 10800

gtatttaaag cctactcagt actaagtagc caactcttac atcttcacct tgcatcattt 10860

acaactggtt cagtgaggta gtacttaaat attactgcgc agtgacagaa ggatatatca 10920

ccctataggc cagggatgat atactatgga tgtacttgga ccttttatgg acttatactc 10980

aatttttttt ttttgtcatg atgttctagt gccaaattaa aatatgtatg caatactgta 11040

tcttctttag ccaaattcat ttattcgaag ttcaacttta ttcatgtgtg atggcatcta 11100

atttttaatg cattaaacct gttagtgctt agaatcaaag ccactgaagc ttttgagctg 11160

caaaggccac ttggctcttg cttggtacag gaccttggct ccatagacta gattagtttt 11220

agctctgtga ggtactcaag gcgctgttga tctgagggct gttaagtggg tcacattcat 11280

agaggtccta gactgtagat aggagggaaa atggagggaa aactctcagg tgccaaaatc 11340

ttcaacaaac atcagtaagt gacccagaga gtgtataact aactctataa cagagacttt 11400

tgtgactgtg tattttaaag ccaaagcagg aattcctttt aaatctgaga tttccagagt 11460

aacaagaatg tgtataaaaa atttattcaa ccaaaattta ttaaatgcca ggcatcgttc 11520

taggaacttt gaatgccttt gtaacaaaac aattatctct acccacatac agcttacatt 11580

ctatttaggg gaatacagat aagaaacaaa caaaaaacat tataaacagt aacttatggc 11640

atactggaac atgattaatt aaatatatga gaaggaagag ctaagtaatg agacttgggt 11700

gttggcaaaa ggtttcctta gaggatttta aaaagaatga atagttatca gagtatgcct 11760

tgatttgatg taatttgctt catgtctttt tggtgtaatt tatttaaaaa ttttagtata 11820

ttaaggaatg ttcagtgttt tataaattgc ttcagatttg ccatgtgtct gctaccaagg 11880

aaaaaaacat ttttgtcata tcaacaactc taaggaatca ttttgtttta gattatttgt 11940

gtttaatagc tttcctgagc aatctgttct gtttcaaaat attttttcag gatgggctgt 12000

cagaaattta aattgtagta ggtgaactaa gttttgacgt gtcataaaag ttgatgtaga 12060

gcagtaaagg atcatagatt tatgaaaaca aggaaatatt ttactttgga ttgtggtacc 12120

taaaagtata taatagaata aagttctaaa gtgtgtttag aagtttagag agagtttagt 12180

tagagaaaaa tatcaggaga aaatttagaa aagtgaaaga atttcagtac ttaactccct 12240

cttagaaatc atcttacatt tttatcaggc acatatattt gagaggaatt attctaaaat 12300

ttttgatagg taccctacta aggaatgtaa gtgatagttt gacagtcccc aaagatgcta 12360

tgcagtgaat agacctaatt caagactgca gctgaatgga atattatttc agttttagtc 12420

tgggttgcat gtcgctgaaa atagaacctg tatttctttg aatttcccca ttctatatcc 12480

ctcctggaat tcttaaggtc agttaagcac agtaaataat cttcgctcag caagcaaccc 12540

aaatcactat cttcctgttc gacgtagcta ataaattcta agcaaactct aaactctttg 12600

tgaagcttcc taatggatgc ctcactgaac tgaagagctt ttggaagcaa aataaaataa 12660

tcatctaaag ttgaatgttt ggggatcaag gttactaggt caacactaaa tttaaaacca 12720

gtttgagaga cattttttaa tggcacttag aaattatggc taaaggtcag gccgctacat 12780

gcgatgttgc tcagatcctt tgatgctaaa ggtctacaac tgacttatgt gtaagatata 12840

ttttttaaaa ccttaactgt gatttaaagt agcaagagac agtgttcagg ccagggaaaa 12900

taaagtccct ctccccagtt ttttcattgg aatggtttga gttcagtaat cttattgaac 12960

ctgaaaaaaa ttattaatag ctgcttccac aaaggcaaat agggaagctt tcccctgtca 13020

attcctggtt aggagtttct gagatatttt gagactgtaa gaaaactgtg gatgaaatcc 13080

ataccaatta gagatctcgt gtatttggaa ccatttaaaa acgctgtgca gggctcaaag 13140

gaattagtaa agcctgctgc agtacagcta cattactaat ctccaaagag aatagatacc 13200

agatgttcac actgcccaaa ggacacctcc gtgttcctga cggttttagt cagactgtcc 13260

tttcatagat gcaaatgcct aaagtcccgc tgggtgatag gactgtgcgg cttctacata 13320

tgaaattacg aagaatcagt ttgaagaaat gaactgtttt tgataggcag gataataatc 13380

aggttaaatg tttatctttg taaaaaaaat cgcctaaagc aaaatagtag aaattcttct 13440

tcgtctattt ttttttttga tcagcctaat aaaaggcatg ttccaactaa gtgataatct 13500

tttaaatcag tccaatgttg ttgttaaatg atgaattatg gtattttctt tattaattca 13560

ttcattcaac aaatgtttcc ctgagacagg cattgaggat ataatagtaa agaaagcaat 13620

catggtctca tggagcttcg tgaaagaagc agccgttaat caaaaactac ccccataaat 13680

aagtgatcac gaaccaccat tgctttgaag aaaaagtact gggtgctgtg aacatgtttg 13740

actgtgggac ccgaccgaga cttaaagctc tagattcctt tttttcatcc atattgagtg 13800

acttaacttt gccaaaaagc gggacagatt accttggtca tttaaaactt agactcatga 13860

ctctcagttt ggtagaccaa aagataccat gaacaaaaca acaagaaact taaagataaa 13920

atgattctag ggatggaatt ttactttagt ctactagtct ttattttttt tctctatttc 13980

accaccctct tctatttatt ttgaaaaaat ttattatagt tgacttacaa tgttctgtca 14040

atttctgctg tacactaagg tgacccaatt atatacacac acacacacac acacacatat 14100

atatatatat atatatatat aagttatctc acattatcct ccatcatgtt ctgatatagt 14160

ttctggatat agtttcccgt gctatgcagc aggatctcat tgcttatcca ctccaaatgc 14220

aatagtttgc atctatgaac cccaaactct cagtccctcc cactccctca agctcccagt 14280

ccatcccact ccctcatctt tctataaata tagctcctat tcagtatctt aaggagtttt 14340

tgagggacat ttaagatttt ttagcacgtc caagtgtttt tcacattact cattttattg 14400

caaagactcc aattggaaaa taattttgta atattttaaa cacattagaa aacagggaaa 14460

agtgggatca gcaggaggga cccccctgct cccagaaccc aaatctttta acgcaaacat 14520

catgatgctc ttggctagcc atcaacctgg cttctcttcc ttccctgcca ggctttgtaa 14580

agagcagtct ttgttcactc tctccacttc acatatgctt ttcaatgcaa cagagttcag 14640

cttccgtcat catcatcaca ttgaaatggc tcttctgaag ttcctgagac ctcctaacca 14700

ttgaattcag caaactcctc tcagatttta cttgacttgt cctttctgaa gatgatgtca 14760

ctgctgttgg ccctctgatc ttaagcagtt ttctattatc tgactctgtc ctgattccct 14820

tggcctcctc cagtttcttg gaatgccctc ttctattctc ctctgctgac tcattcttca 14880

gctcacctgt taaataccta tctggttctc cgctgccacg atattaacct tcttctatag 14940

ctcctcatcc ataacctata ttttctatgc tctgctgact ttcacatctg tatctgcagc 15000

cagggtctga cttctgaccc acataaacct ctgcctccta agtggttcca cactacactg 15060

attttattat ttcccctgaa aacatgtatg acgagcatgg tcccagtcat tgtacagctg 15120

gaaacttagt cataatccga ccctgccctt ctccttcact ccctaccagg tactacggtc 15180

actgcaatca gtgaattgta ctctctcaat ctctctacag tttctcccct gctccccagt 15240

cccattgcag ttgtcttcat gtaaaacttc gtcacatctg ttcctgtgca gctgtcatct 15300

gatagatctg ctttcgtcct tcctctcatc cctcaccctt cagcctgctt ccagagcatc 15360

ttcccaaaac atgagtctga tcatgttact agtctgctta acagccttta ggtggtcccc 15420

atcatcatgc ctttcaaaat gttcctggcc gtggtgccac tgctgtttag aactactgca 15480

ttacttgaca acaagaagaa aagcagggta aactagtagc caaaggaaaa aaaaattaat 15540

acttgaatgg tacattgcac ttaaactttc ttagcactta ttatcttgga gtttttagag 15600

tctatagagc catgctaatt tttatatata ttctaaactc cctgcctttc aagataagta 15660

ttttaagtac catttttact tgaggtactg cttgttaaag ctgaatgcag catggggaaa 15720

atgtttgcca aaaccagtga ttttcttttt gtcataatta ctgttaaact ggagttcctg 15780

tagtggcgca gcggaaacga atgcgactag gaaccatgag gtggcgattt ccatccctgt 15840

tcttgctcag tgggttaagg atccggcgtt gccatgagct gtggtgtagg ttgcagacgt 15900

ggctgggatc ccatgttgct gtggctgtgg ttgtaggctg gtggcaacag ctctgattag 15960

accctcgcct gggaacctcc atatgccgcg ggtgcagccg taaaaagcaa aaagaccaaa 16020

aataataata ataataataa tcactgttaa acaatttcct tgactggaat agtttacctc 16080

gagactactg cttgtttgca atattgaaat cgtgtcatca gcatgtgggg gcacttgttc 16140

ctatatttgc aatggcatgg ggtagaaata caataaattc gcaaaccgta tcaagcaaat 16200

ttaaagttgc aaagtggtct aagaaatttc tttctaccaa accggtcagg gaggagactg 16260

tcctgacttg ccttgattct aactcagatg gcattacaga tgtcatgaat aagtgctcat 16320

ctcctacagc tagcccataa tcacatttta gccagagtca aatctgagtc agttaaagct 16380

actaaacctt acattgattt tggtattttt atcagtaagg taaaaaataa tctaattgtg 16440

gttagtagaa aaattatgat gcctgaaatc tgttgcttct tcccataaaa tattagatca 16500

agacactaaa atctgcagtt tattaaagag aaatcttggc ctcttgttca agatgataga 16560

ctcagcatag gtatttgttt tcctctctcc caagaggcac atttaaggtg agaataggga 16620

aaataagcag aaagaagagg aagaggaaga gttaaaatgg agaagaaatc attgcaaaga 16680

gaatagactg ggggaggggg tcttgagcag acaagaaact ttaaacggtt ttttccaagg 16740

gaaaaaaaaa atgggtggaa ctgcgttaat gggtacgact gactaaagaa agatgcagtg 16800

ccaaatttgt accagagaag ttgtggagat gcccagcagt attgtcaaga gccaaatggg 16860

ggcctgacaa caaggtgttc aagttgaagt cttaaggaca gctagaccag ttttacttct 16920

ttctcacttt catatttaga atgatagatc atcaattact cacatggaaa aacaaacaaa 16980

caaacaaaca aggaagactg ctctcctaag acgttaaaca gcctgttcag gggaattgcg 17040

gctgctgttg aggtgtgctc tagagtaaac cattccccag tcaagtattc agggaaccct 17100

aggaccaacc cttttccatc agcaaatgat gcctgtttcc agagctccca gccagctttt 17160

ctgcccagag ttaagactag taaagaaaca aactcatcct gaaagcaaag gaagctgtgt 17220

agctgcatag gataacagac ccacacctcc gttcttacat agaaatggcc aaagaccacc 17280

acagatatac acagaaaacc aaaagcatga aaaacaaaag ccaagataaa gagaacagtt 17340

gccccagagg aagcagagat gtttgagagg ataatagaaa aactaaaata aaacaactct 17400

ctaactcata ttcccagaga tatttgaaat acttcaatta taaaataaga aaatgatgct 17460

cttaaaaaaa atcacaaaac accaaaataa acaaaataac tagaaaccag gagtaacgtg 17520

aaaaaataag ttgacaaaag ataactatga tttgatatca tttatgtaaa ttattgaaga 17580

tttaaaacat ggtatgttat ttatggatat atacatgtag aaaatgaatc aacacaagtg 17640

taggagtggt acatagacac ttaacaacac caggatatga aggagtagga gagaaagtag 17700

ggggcagttc tggctgcaac tatattattt tattttgtta gaaaaagaga aaagctgaag 17760

aaaatatttt ttaaaagtta ttatctatca aatctgggta gttagtgtct attgaatcac 17820

agatgcctat tctgtatact tttgcatttg tttgaaatat ttgtaattaa aaattcaaaa 17880

attgaaatac tgaattaaag aaaaatttgg aataattata taataaaatc atggtctaaa 17940

caaaaaaaaa atgaaagaaa aacatgacat gcctcttaac atgtgataca atgtaaaaaa 18000

ggaaaatctg gttttaggca gcctgagatt atgataacgg gactgttgga gaaggaaata 18060

caatcccagc actccttagc cctgcagtaa taaatgcata tatagctgtt aaaatataaa 18120

ggctatctat gcatttttag ctcttaaaaa atcaatctgt agaaaagcat gcaaaggagt 18180

gcccattgtg attatgtcac tcaaggatct gccattatct ctgtgaggat gctgtttcga 18240

tccctggcct cattcagtgg gttaaagatc cagcattgct gcaatctgtg gcataggtca 18300

cagatgcagc tcagatccag tgttgccatg gctgtaggcc gcagctgcag ctccaatttg 18360

acccctggcc caggaacttc tatatgctgt gggtgcagcc ttaaaaagga aaaaaaaaaa 18420

aaaaaaaaaa aaaaaaagaa gtaaaaagta aaagaaaagc aggcttaatt aagactggac 18480

attggaatgg gggaaagagg aatcagaggg aggatcaggt cactaaagcc atgttacaga 18540

gggagagtca aatggtataa gctatccgaa agtctcacag tccttcaatc atcagatcgg 18600

tttttgcaac atctctatgt caaaggcact gttttatgag attcgcaata cagtggtaaa 18660

caaaacaaac tccccactca tgttaaattt acactctaga tttagaagca gacattacca 18720

aacaaggttg taaaagaatt agaaataata cagctattga aaattggaag aggaaaggga 18780

gggcagatta atataacttt gaatactaca tcttcattat tcacaaggaa gaatctattt 18840

atagtttcta aacagcactg tttaatggta aaaatgtttt gtatctgtgg aaatgatggt 18900

ggaaatattc attacctgta tctgtattgt ctaataccat aaccattaac tgtatgtaac 18960

tgtggaaaac tcgaaatgtg gctagtgaga ctaaggaact gaaattttaa tactttagtt 19020

taatccattt aaattgtgac gtgtgactgg tagctactgt actgaacaat acagattaaa 19080

atgaagcccc agataaaaat tagaaacctc aataaattag ctacatgaac caccagtaga 19140

gaacatagat gaaactaatc atgatcgaga attcattgaa ttaaaatgta attagaaata 19200

ttttgttatc ttcagattca aaataagttt gtgcacaata aagaccattt aggcttgggt 19260

gatgttttat gtatgtattt atttatttat ttagtctttt ttaggcccgt acccagtggc 19320

agatggaagt tcccaggcta ggggtccaat tggagctaca gctgccagcc tgcaccacag 19380

ccacagcacc accagatctg agccgtatct aagacctaca ccacagctca ctacaatgcc 19440

agatccttaa cccattgagc gaggccaggg atcgaaccct catcctcata gatactagtt 19500

gggttcatta acaactgagc caagatggga actccttggg tgatggattt tttttttttt 19560

tttttttttc ctttttgcca tttcttgggc tgctcccacg gcatatggtg tttcccaggc 19620

taggggtcga atcggagctg tagccaccag cctacaccag agccacagca acacaggatc 19680

tgagccgtgt ctgcaaccta caccacagct cacggcaact ccggatcctt aacccactga 19740

gcatgatcag ggattgaacc cacgacctca cggttcctag tcagatttgt taaccactgc 19800

gccacgacgg gaactccagg tgatatttta ataatgatat atgatgtata tatatgagcc 19860

tggttttggt gagacaagct ggaggtttgc agatctacag ttttttgtgg atagtaattc 19920

tcacagctct acaattctag gaccatattt caaaatttgt agcttttagt ttctctgaaa 19980

ccagcatctg ttaacctcac tgactgatat ttaccttatc tcaagccaaa taataaaact 20040

ctttatgaag agcccctgct aacaagaatt gtttgaagct tagagctctg attcttgtcc 20100

aattattctc tttaggtgac acttcaagta taacttttat ctcattttta aaaggatgaa 20160

tgtctatttc ctctgaaaga cccaaaagtt tcagtagtaa tcaaccaaat acatgcatgt 20220

agtagaaaat caaacagtgc aagataactg aattaaatgt tccctttcaa ttctccccac 20280

ctccgcagat gatcttgttt aacaggtttt ccatttaatt ctaaaaaaca tgcatgtgct 20340

agggagggat atggaggaaa ggaagcagta tagtagtcct tgttaaatta agagtaagaa 20400

aggagggtca ctttttctag gggtggtgca gatggaattg gctttaactt gtttataata 20460

ctttgctagt tttgaacctg caacttgttt ttgatgattt ttgaaaggaa ggagaaaaag 20520

tttgggatag ctgagaaagt ttttcacaga aggcagttga tctgaaacat ttaaatttgt 20580

gaacagaaca ctacttttgg agagctgtgt catatagact cagagcagct ttattacact 20640

acattatgag tgaggaggaa taattctttc tattgcaatc taaaaaaatc ctctggtgta 20700

catggtagca tgtcccaagg atctactaat aaaaagctaa taataaaact cttatatttg 20760

agatgtaaat gaacataccc tggatttgct tcacatgata gtagcatata ccacgtatac 20820

caagttatgt ctgcagatta atttggtctg atggttggat aagcagaact tccatattcc 20880

aacttgccag cattagttac agtacctgta gttattttgc tatttagaga tgaaacttct 20940

ttgagaggag aacctattta tttatatttt tatttattta attttttttt tttttttgcc 21000

ttttctagga ctgctcccga ggcatatgga ggttctcagg ctaggggttg aatcggagct 21060

gtagctgccg gcctacgcca gagccacagc atcgcggatc cgagccgtgt ctgcgaccta 21120

caccacagct cacagcaacg ccaaatcctt gacacactga acaaggccag ggatcaaacc 21180

cgcaacttca tggttccaag tcagattcgt taaccactga gccacaaggg gaactctgaa 21240

cctatttatt gaatgtagga tattcttgat gatagtaata ggctaatatc gaaaaggtat 21300

aatttatgtt attggaaata aaatgataat tatcccatat cccacctcat caaatagaaa 21360

caaatggatg ttttgggcat ggggtgagtc aactctgttt atgcaggttt tcgcatgact 21420

tttcttttaa aaaagtgtat ggaatgcatc caaaacaagt aatattttgt tagatgagaa 21480

gaccaaaagg gaaatcctag agaaaggagg atggatgatc ttgctttaat gtcaaaactg 21540

acaaagacac cccccccttt gctcaagata attttctcaa caagacccag attgcttcag 21600

tctttgccta tcaccctttg aagcactttc tttattatta ggagtggggg cagggtttcc 21660

tttgaggatt tttacagcag tgaagagttc ctcctttaaa tactttcaca tgatgaggga 21720

attccgcttt caaagtgctg tcatcactgt ggcttctctg ccataagacg atttattctt 21780

aggtgaaggt gcttcttaat ttgattgtca ataggactta cctttctgat ggattctctt 21840

tctgtggagc ttggcttttc tgagagacct agcaaaagag ctcaacattc gtttttgtta 21900

accgtattgg tgaggttgac tatgattctc ttttgaaaag tgtttcatca cttacagtta 21960

tagacgtcag catggctcct ccttatcagc cttcgccagt tgagatgtgg gtggcatgtc 22020

cagtattcca tcttgaaaac tgcaattctt ctaccgtgca gagggacacc atgcatttgg 22080

ggatgtttgg tgtctcttcc aatgtttaaa taacaccatg gtaaacaaca aatacaatat 22140

tcaagcatca agagttttcc cagaaacatt gaaaggtttg gagaaccttt actcgtgtca 22200

ttccacttca tcctgctcaa cttttgaata tagtcttctg gaaaattcta taagtattta 22260

attcatggag gcaaaatgta tagctgtatg ctttcatagc tcatttgaaa gtagttcctt 22320

cccctccccc aaatttttag taaaatgatt agtaagcaat gattcgcagt tattaggata 22380

gaattagaat tcggaaaccc caaagagaat tgaccatttt ctcaatatta ttgccgaggg 22440

gtaagctcct aagtctttgt ggttgtttta gataaaaaaa taaaaaaata actcctctat 22500

gtgccaaact agattacatc atttacatca ttgccgttgg actttccctt tgaacagttt 22560

tctttagaaa gtgagagaat ctgtgttgtt tagggtacat tcattatctg cttccaccta 22620

gaggccctta agtgcaacag tatacttatc aactaatcca gactggcagg aaacatattt 22680

cttaaaacct acgacttcct tcctgctaga gtgttcaaat gattgacatc tccgcctttc 22740

attgaggaag aaggacgtgg aacataagac caggggcctt aaactaaatc agaatattca 22800

gattctagtt tttactctcc tatttcctgc ctgagtaact tttggcaagt aacttatctt 22860

tctgctcccc cattcctctc cactacctcc caaaacctct gtaaagtgag caacctaaga 22920

tgatcatcat gcgtttttac cactttaaat aaaaagttta cttcacctac atagatgggc 22980

ttaattatag taattgagcc aaatcaggtg gtccagtgaa aaattagcat tgatattcct 23040

taatctggtc tccaaagagg aatataattg tactctttct cctgtgcaac tctgtgatgt 23100

actgtcatta ttcaccccac tttcttattt aaagcttgtt gaaaatcaga ttaataaaca 23160

tttgagctct gcctctgtgc tgtataccat ggtagaaatt ctcattacct aagcaaggaa 23220

gaaagcatgc aaacaaatca caaatctagc gtgataagta gaacaaggta aatatacata 23280

agatgtagag gcagcgtagg gagtatatac atattcctct gtacctattt taaattctag 23340

aagtgggaaa atgtgtatat ggtagaggga ggtagcttta tggtaaattt ttaaaaaacc 23400

aaaacctcct aactatttac ctgtttacta ctgaatcaaa tatgttccct tgtaatatta 23460

gttgctataa actgtagtca aatgaattat gacatgaaag attaataagg caagatctta 23520

tattttcttt tcatagataa attatttttc aatgtcttac aacatcacca ccaaacttct 23580

ctttagcaat tgaaatattt gtcattgaac aaggaagaac acctaaatgt cgcaactaat 23640

cattaaaaac caaatactat agcctggaat atattcaaaa tctctttgaa atgtagacat 23700

gggagaaatg agttttctta aggatactga tatttcttag cataattaag ttttgtggaa 23760

tattatctag tatcaacaaa gttaaggttt tcctggtgat ttgtagcaat cttgaattca 23820

aatcaaggaa ttgaaaagtg cgtaaacaat ggggtgtgaa atctcgcact atgaaagcta 23880

tacttttgca tggtggagga tgcttgggga agtttagcat aaaaacttga cctgagctaa 23940

tttgttttgt ttgaaactgg aaatcatatt gttgaaattt gcattctgtt ttctttcaag 24000

aggtgtattt tcataaagct ctctgtcttt aaagagcttc ctcctttctg caggaaagat 24060

ttggcttctg aaaaagaaaa aaaaatgtaa aagcagttcg gcaacattgc acctctttca 24120

tcaatttggg gatgcaggtt tcagcacagt aactccagac atttttctgg aaacatgtaa 24180

aagcatgccc aaaagtactg aatattataa aacccataaa cattgtacac ccataaatat 24240

gtgtacaacc agagtttctc aaaactgtgt acctatttta aaaatcacta gaataaatga 24300

aaattggaac aagatgctat ttgagagaag gtgctaccta gagatgctat atgactctcc 24360

tatgaagtga gcaagatgcc catcaataga agctatgctt tccagataac ggctctcaag 24420

aaagtagata aagatggaaa caacagcatt taaaaaaaat atattttaac catccaaact 24480

actgaaaagg cctgcttcct aaagagacat ggaatagttg aaaagttaat agccagatca 24540

gttggactgt gttttgggaa acatcacatt gtaagattca tagtgctcag gactgtgccc 24600

agtgactctg aagataagag aaagatgaga aagtggtgag aaagagcttt ataagttcaa 24660

ggacgctcta aaaaactcct ggtctcatga aacttaaaat cccaatgtaa ctgtaatctc 24720

agtatgagta aaagtttatt cttaaagata gttgcactct cgcaaaatat attgcctttt 24780

ggtctttgac tttttttttt ttcagattga aaaagttaag catgattcat actctaaaga 24840

tttccattct atgcttagta tttcttttaa aattgcattc taacccaagg ctgtgtatct 24900

cctctgtttt ggttcttttg cctgctgtaa cccaggttgg ccttcattcc atgatgtgat 24960

cagttctgag gcagtcacat tcacagacga cttttcctac gggatgcaca gggtggaaac 25020

cagctgctct caggtcagtt aacccacctg aaaaccccaa atactgctca ctctgcattg 25080

caaacacttt taaggctcct ggttgtgcta aatatagcaa ctaaggccaa gtgacccatc 25140

acaaaaggga cagacaagca caatgaagtc tctaccaagt cctaatgata ttgcagtctt 25200

gaggagttta gcaagcccaa ttacatgtca gtcattggct tcaacactat taaacaaagt 25260

cattggtaag aaaaatcatc ttcaattgac tcaatacagt ggagaacagg ccattcttag 25320

gggatgtgaa gatatcatta tgtggcctta cataatctga cggttaggta atccccctct 25380

ccctgcactt ctgtgcactc attgttcagc cataatacct ctaatggtaa tgatccaaac 25440

acggtcaaca aagaccccag aaacagctct ccatgggtgg atgtaaggag ggtgtttgga 25500

gaacggcagg agaggaagtt cagtggcgtc tgtgtgaggc agagagggaa tgcgtgatga 25560

ggtgattcct tctctgctcc tagaactagg cccatcctcc tttccaagga tgttctcatt 25620

cgttccttcc gctctactct aaccaaaata gtgctggatc taagggggag aatgaacacg 25680

tgttgaggag gggtcaaaaa tacagttgcc tgtcatccca gtcaagagac ttgaattttg 25740

tgtttaaacg cagagacata ccagaacttt cttgcacaca gacttagggc aagtgctcat 25800

tctattctag agtaggtagt cctgttctaa attgtttgga ataagtgaaa cataaagtct 25860

tagtattgcg atgctgaaac tcattgatac tacggagtat tttgtcaggg gatgtctgtt 25920

gtgcaggtca gaggtaaagg aggcacaatc aggcagcagt gagagcaaag ggattgccac 25980

atgggcctcc ctcggcattc ctccacctgc cctttcacct ccttcatcag gcacaagagt 26040

tttagggagt aagtggcata tttactacca agtagggagg gagattggtt tgaacctaac 26100

catctgatga atttctgttt tcttttattt tttaaaaata agactggaat ttaactgtct 26160

tccacgtatg agaaccactc tctcccttga aaactggact ttatatttta cctaagtcac 26220

atttttggtt cactctggtt tctcagatat gaattttagc aagagagtgg taccttttat 26280

ttatttattt ttcttttttg catgtggaac tttccagcca gggatcaaac ctgtgccaca 26340

gcagtgacaa tgctggaacc tttaactgct cccaaaacgg tggtactttt tgatgctggg 26400

tcttactctc agagatttta attaattggt ctgggtgagg cctgaacatc aacattctaa 26460

aaagccgccc tggtgaatgt tcacaataac cttgtgagta gatattgtca ttatctctca 26520

ctttacagtt gagggaactg aggcacaggg agtgcaatta cactcgcgta aatattcaca 26580

gatttaagtg gcagaactga gacttgaatt tggactttaa ctccagagct ctttgcggtg 26640

ctcagataga taggtagata ggtaggcagg taggtgagta agtagataga cagatagata 26700

catgccccat aaagagtcat atcgaaataa gagaaaaaaa aaaaaaaaaa caaatcttat 26760

gatcccgcag cagttagttg aattcaattc agtttgggac tcaattgaat ttttattgat 26820

tcaaaaaaat aataactacc gcaaaaacaa attgatacta tgttctcagg agttcccgtc 26880

atggctcagc agagttccca cagtggctca gccaaaatga atctgactag catccatgag 26940

gacacaggtt tgaccctcgg cctcgctcag tgggttaagg atccactgtt gctgtggctg 27000

tggcgtaggc ccatggctac aagctccaat tccaccccta gccaaaaaaa aaaaaaaaga 27060

aagaaaaaga aagaaatgat ctactcagga atctaagata cttgacaaaa aggtaactgg 27120

ataggatggt acatagtaat ttgtaactta ggtacatagt acttagtaaa cttctaggac 27180

tactggttgt tttttttctt tctttttttt aagctgcctt aaaggctatt tgtcacatga 27240

ctcctacatt ccttttcacc gaaaacccac tttcaaacag tgagttctat gaaaaccaaa 27300

taaaaaaggg acattaaaaa gtgaggtata agatacctgc tttccttcag gccctgctct 27360

ctgatgcctt tgggacaaat tcttgaaatt cacactttga atacacttgc tcctttctct 27420

atgcccgcca ctgaggtaac atcagtagga gttcagacaa gatcatttca aaggcgtcct 27480

acggtgacca aagaggaaca caacccacaa accgtgccac aactaggcaa ccctccatag 27540

atatcatgct aacctaggca taactctggg ccctttggtg aaagccctag gtgactgttg 27600

tttattcttt gggctttttc taacaattgg ataaagaact aacactgact gacctattcc 27660

ttttcagctt cttttcccct caactttatt tttcacattt agaaagcatt tgacccattc 27720

ttcaaagact taaatttaaa gagaagaggg aggaaacttt gttcctttaa cgtcatgtgt 27780

aaaaatacat gctttgagaa attgagtacc atgtagaaaa taattgggtt ctggccccaa 27840

agactgtgtt tgctggaacc tccccaagtg gacagagtga gcgcgtctgc cacacagtat 27900

acgaaaaagc ccactaaacc gaagtctgct gggtgtcttt cctttcagtt tcttttcttt 27960

taatttatgt gtttggcaca tttgctatcc tttcgtttta catttttctt tagttttctc 28020

ttttccatgt tctcattcat cctttgtatc ttgacttggg aaaaaaactt agtggttccc 28080

ataactagag taaccatttg tatttcactt cctagtttct catccttaac actatttttc 28140

attatcagtt tatcttcttt tacctctgtc ttttcattca tgtattctct tcacctactg 28200

gccacttctt tctctctctt tttttttttt tttttttttt tttaactttt ggatggcgac 28260

atggtgtggg aaacttctct gatttgtttt ttctctttct aaaattcttg ttatattttg 28320

aattcacatg gcttgtcctt tctctggact gaattgcatc agaggggttt gtgttatgaa 28380

gactctctta gatgctttta gtatcttaga gacattctca gttggacgtg ttatgtgggg 28440

tgcttctttc cttgtttacc acaaggagca tctaagcgtg aaaacccttt tattgccagt 28500

ttacacaaat aaaccaagct tgcatggcag aaaccagtag attccatctg cctgcctgcc 28560

tggcaatatg gcaatatttt ttttttgtct ccgggtctct aactcttgct ttagtcctta 28620

agattcttct tctccttccc cttcccctcc cctctcccct tctccctccc gcctcttctt 28680

ctttccttct ccttctctcc ttctctgccc cccaccccca tccccttctt ctcctcccct 28740

gctccttctc cttcaccgtg atctggatca taccatgcag tgaaataggc ctcctatttg 28800

ggtaggagtt gtaaacatta ggagcaatac ctagcacccc tctacactac actgtaattt 28860

ttccatatgt ctaatgactt cttaggtagt aatattttag taaaaatttt aattgagaaa 28920

gttaagattt attcagtact ataatcactt ttcagagaaa tattgcatca ggtccttcca 28980

aaaagatctc agtgtgagaa acccagctca ctgcctgatc tttaagagaa cagcacagaa 29040

aacaagcaag ggtggcagag attcttccct gccttcatca ggattcctgc ttgtggtgga 29100

aactggctac cctgttacat tctgatgctt cttgcaagta tatttgtttt cttttactat 29160

tctagttact gagttgagaa attatattac agctatctct tccaaagagg atctttttat 29220

cttaaatttg cccattttcc tgagcaattg tatcttcccc tggctatagc gatttcctaa 29280

caattttcag ctatagagaa ttttcctggc attcccattg tggctcagca ggttaagaac 29340

ccgacatagt ctccgggaag atgtggattc aatccctggc attgctcagt gggttaagga 29400

tctggtgttg ccgcaagctg tggcataggt cacagatgag gcttggatct ggtgttgctg 29460

tggccgtggt gtagaccagc atctgtagct ctgatttgac cgctagcctg gaacttccat 29520

atgctgcagg tgtagcccta aaaagaaaaa aaaaattttt ttccttaatt gtatttaatt 29580

tacatttttt gttagttgta aaatatactg cttctattcc agtttcactg actactatga 29640

ataattgctt gacagcttct ccctaatcct tcaaccccgc ctgttaagca acacgttgtc 29700

aggaataaat tcagtgcaaa gaggtaagat ttgcctggtt aatgctgtag ttaagcaaag 29760

taagtccttc ctggaaatac ttggggaatt cttctttctt tagatcaaac ccctacatct 29820

gccgcaaatg ggagaccaat aaatcactta acttttgtaa taatctttaa ggaaaccatc 29880

agaaccctga gtttacccaa gttggactct gtttatgact tgtagctatt actgtttgaa 29940

tctatggtcg agtcccttaa gtttgtttta gatcattatt gatttgttct aagtcagaca 30000

tatcaacaga aaggtaaata cccacccata caaagtattt tctcaagagt tacgcagttt 30060

tgggtaaaac cccaaacctt attaatcttt tgcagatgac aactcatata ttgctgatga 30120

ttaaagatac ccatgtgtta caatatggtt ccagactcac agcccatctc tctttttctt 30180

gatctttcag taaaatcata atgctaaact ggtgacttca caatcatttc cttggcaaaa 30240

gattaatatc acaagcacag aggaaaagag ccctacttac acacaagtgt cttcaataac 30300

ggtgggagaa aacaggatac tactggcaaa gtaaattgct tttcttctta aagctctttt 30360

ctccttccca tcctgcccca tgtgcccaag tgtaaaagcc ctgctttgaa actgcttcat 30420

cttgtagatc aaacacttaa gagaaaatga caataataaa gccccaaaca tcctaggaat 30480

agctgggaga aaaccctcta ggacaattgg aaatacctca gtgcagaaat atgtaactgt 30540

gtttaataca ctgtgtttct tatgcacata attgagaatt gcagtcagat tttaaggaat 30600

tacattaaat gtgacgctca atcacttttt tttttttttt tttttttttt tttttttttt 30660

ttttgagagg gtcctttccc ttggtttccc gcaggttctt tatttgtcag cttttctgag 30720

tggccagctt agatgataaa ggattcctat gggaatttac aatttttctt ttgaacaatg 30780

ctcatctcat gcttcttgag aatgagagct agaatgtttg gtcatggttt gtattttacc 30840

tgtaaaagaa ggaaatacat aaccacttta aaacaacttt caaaatgcac accaaagatt 30900

ctaccctgca ataaatcatt aacagccggt ttcagagatt tcttcatcgg gcaaccagtt 30960

tgtttcagtt acaaccaggt gtgccgagag atcccaacac attgccagtt ggctggggcc 31020

ggggaggcag agctcgaagg cggctaaaga aacaatgcat cactgacaac tttcattcat 31080

catcccagcc ttccgccagg gctagaagct ggaaagaatg ggctccatgt gcgttcccct 31140

cagggaggag ccttgacggc ctgcgcactt ttaaaagaaa ccgggtactg atcttttatg 31200

ggggactggg tgtacctggg aggaggcaga catgctgctg tggggttagg tttccttcag 31260

acaaatggcc ttgccacttt tcgggcaacg gtgggcatct atgtcgtggc tcagatacat 31320

gaacacctta ccacgtcttg aaatattgct cctccctcag ttctctgagc ctggtgtgaa 31380

tcttccagaa caccttctgc aagcaatacg aaacgtagca ctgttgttct taggggaccc 31440

acaccttttc tgagggtttc cctaccccgt cactaaccca tagagttgtt tgcaaatgag 31500

gtttgttgcc ttttcccaac tataatttat cttcctcatc cccctctcgt tttcttttgc 31560

ctggattaag gacatctaga gtctgtgttt ttctttctgt tttaaccgaa agactgtttt 31620

cagatttagc aaatgaaacc gaagattaat aaccaattca gtccaacaga ttaaattact 31680

ttaatttgga tttttaaaaa acgatttggg aaataaacac cactaaatgg cttcaagtat 31740

gctgttacta tgctttattt ttggagtaag ataaacagaa ctgctttcta agtatgcttg 31800

agatgattga taagggcaat atcatttatt tttgcccaga tagctgtatt ctctcattaa 31860

gaacaaaaca aaacaaaaaa acaccaccaa aaaacctatc tgaggctttt atgtgcaaac 31920

tccatctctc agatgccaag tgatcaaagt gccggaactg cacgtgagat caataattgt 31980

gcacacatac tggtcactga gagcaccagt tcctgttggc ttgcacgctg actgttttgg 32040

caggcagaaa taggtgcttt cctgtgtcca ttgggtaagc atatgtattt ttgctggtgc 32100

atcttagctg ctgatgcctg aaaatttggc ctcaattgtt tatagaaagg aaaagcaatc 32160

tgagctgtca cccttgatat tacctaaacc ggcataaaat gtggtaaaat aatgcaagat 32220

ttgtttttta ggcctgtttc atattttcca ggcaatagat caaatcggtt cagggttcat 32280

ttttagttct ttaataagct gacacagtat gttggcagta aaataagaat gcacttttaa 32340

catggaagtg ccatatagtc aaagttgcta tttgtgctat ttattaataa cgagagaacc 32400

ctcgagaact gcgtttagca ctttcagtgg ggaaactgag attctagtct ttccattgta 32460

atgagcacca tccctcactt ctttctcctc atctccattt ggtgtggagg gacagaaaaa 32520

ccccattccc ttgtcaattc ctgctttttc tgcaatttat gtgttttata catcaaacaa 32580

gcacagtgag aattaataaa tcatctgtat gcctcttctc tcatgttctt ccgtaagtct 32640

taactcctcc atctgtttga aatggtagat acctactcct tcacttcctc caggctctta 32700

tcccaaagtc atgcagggaa caccacctga ctaccagctt tcctcaaatg tgagccccca 32760

agctgaccat tgatctccct ctctttcttt ttgcttctta gcatttgttt ccccctatta 32820

tacgttacag ttccttcctt cctttgcatt gtctgtctcc cccatgaaga tgtgagcgcc 32880

attagccatt gcctgtacct cccaagtctg ggacagtgcc tggcatgtag caaaaacacc 32940

ataaatattt gtgaaatgag tgaagggggt catttgcccc aggctgtggt ttctctgcca 33000

catgatgaac gatgcgtttt cacagcctgt ttgaatgtgg tgctctactg tctccatagc 33060

cttcaccccc gttcccgctg agaaagggtg ggaaataggt aagtgttgac ctgcactgtg 33120

tctcctggag ccgtagcagc agactggttc gtgcggggct ctgttcggtg agccaggttt 33180

ttttggagac acattctcta ggctggattt ctcttaaata ccatcataat caaccatctc 33240

tccttttttc tctgtcagga ctctctgcct atccccagct ttttggatag tcagtgggta 33300

cagattgagc ttgaaagctt attccttctc cacaagatcc agtttgagcg ggtaaaggag 33360

cccgcgtggc actccaccta cgggaagcca gaaaaaaaaa cgattgtgtg gactagggat 33420

gtttccattt gccacactgc aagactatag tcccggtgac cgtgacataa atctcttgag 33480

ccttgccatt gctttgctgg gttccaagaa catccagtgc ctgtctgatg ttaagtttta 33540

ctgcccatga aagcaaacat cacctccgtc atggccagtt aaacagtatc ttttacaggg 33600

gacctctgca tatctgaaag ccacccatcc aaagccaaaa gcaatggggg tggggggcgg 33660

gtagaaattt tagtctgcca tgtttcttac aacgggagat gaagggtcaa ctcaattcat 33720

aaaattactg cagtcatttt ttttcagctg gccgagaggc ttcacagatc aaaaaataca 33780

gtggctccta aatgattttt gatatgcaca attctggtat ttttttttct tctcagcatg 33840

tggtgccaag cttgccagaa accatccttg cagctattga caggcagatt agaaaccatc 33900

caccgtggtt tccatcgttc ctctacaggc cgctcatcca tttgaaccgc tgagtgtgag 33960

ctccccacca agtctgtttt gtacacgctt tgttcgctta ctttgtgctc ttctcttgca 34020

gtgcggtgct cacctcgggc acatttttga cgacggacct cgtccaaccg gcaagagata 34080

ctgcatcaac tcagcgtccc tgtctttcac gcctgcggag agcagcagcg caggtggcag 34140

cgagggggac ggccaggccc agaccaatag ggccgagctg tagaggccgg agagccggcg 34200

gacaagtgcg cttaaggcgc acctaccatg acaaacccag tatcctctat ctcagctcta 34260

ttttttcaaa agtaggaggg cagttttatg ctattgatat ttttcttctt ctgctaaaca 34320

gatgccctag ccatcagtgc attttgctat tgactagatc gggaactgtt cataccttta 34380

gtgacccttg gagagagctc tgtgaggctt cttcaccagc cactcaacta ctgcttggca 34440

ttattttctt tgcccacata gcttcttttt tgttgttgtt gttgttgttg ttctccttgc 34500

ttcagaggca gaaggtgctt ggccttcaaa cataaacatc aagatctcct ctgcagtgcg 34560

ggacccaggg ctgggaaggc agggctttga gacctgggag acatatggcc gcgaggcatt 34620

tgtgcagatc cacctaagat aaggatggag agatgtctta tgagacttct tagctttgtg 34680

gaaagtttga gctaaggtcg tttattctca ctaaaagggt gtgaaggtct aaagtctttc 34740

cttatgttaa attgttgcca gatctgagag ggcgcactga gtgttgtggc agagaagtaa 34800

acattacctc atggttaggc ccttatttta ttttattttt cattgaaaac attggaatgt 34860

aaggcaactg taaaaggttg agccccgttt tgtccaaagt tcacacagaa cacctgccat 34920

ggccttttat aagtctgtcc ctccacacct aataccagat gcagctagca gggcattttg 34980

gtgaattcct tgtgtaaaat aaaatatttg ggacagcaca cttcatcaag gtagcagacg 35040

taagagagag cagagaaggc caaggaggaa gcctgggtaa ctagatgtcc agtgctctgc 35100

agaagggctg acaagatacc acaggtaaag aacggagccc ggagagaagg ggccagggac 35160

ctggtgcaca gggctggcca gatgacctct ggaaggacac tgggggttaa aatagacata 35220

gcaggagctc atgtcccacc cttcccgctc atctgctctg taacaaggga caaactattt 35280

agcttactct ttatgtatat aacacagact taacaatatc tgtaaaataa ttgacactct 35340

caggggaata ttactatatt gctatacaat tatgatcagt tttgtaaaaa ttaactgatc 35400

tggttctaat tgcctcaaag gccgaagccc agacatttga aatggaaaga aacagagagg 35460

aggttaactt agctgattgg tatggaaaca gttgggccaa gagccagaaa attttctttg 35520

tagcaacatg gctggtttta ctctaagaag ctctgctggt ggaagtataa catttagtct 35580

ggtgggctgg atgccattgt gcacagtaag gtttttatag ctccaagcag gggtaacatt 35640

ccttggtatt gctgcagcca ggtgtgaagg ttatgactga tatatattgt gtttgtaacc 35700

ttgaatgcta atcctgaaat gtagtttttt aaacatattt ttgtaaatct ttgaaagctt 35760

atgaaaaccg atgtctttcc ttgtaagcac caagaccaaa ataaggaaga gggacacttg 35820

acaatcttaa agaagtaatg agaaactaaa atgaaaagtt gcgatcagtc agaatgttgg 35880

tatggtcacg tccatatcct tctgtgtctc tccagatact tcctggtgag gcgctgacat 35940

cttggtgact cgtcattgtc acatgggcca cacggaactt tgcacttcca aatacattgt 36000

gctcttcctc tttcagctcc cagatgtttc acctcaggct ctggggtcac tctggaattc 36060

gcaggtttca cttccttcta gaagtgagat gtagaattta agatccttga agcctcagca 36120

tccagactat cccagaagtc tttctcaacc ttgcagaggg tgtcctgaca ttgcctagta 36180

acctgaggat ccctaagagg tgggtccccc aggacacgcc tgtccgagca tgcagtgcat 36240

ccttcagggg ctcacacagt acttctcgtg gctccagctc taggtggtgg cctgccccag 36300

ggtttgctgg actcgagcac agttttgggg tggacaagga agataaacgg tggccttctc 36360

cctctcccca gagactctag attctcattg ttcctccagt tttattgtag ggcttaacat 36420

tcccctcaaa aagctcctct tctaatgctt actctttgtt tcaaggaaag cctgtttttc 36480

ttctaccaca ttttccagca tcaactttaa gaaaaatgca acatccattg aaaaaaagaa 36540

agtggggtgt atgcacatgg tttaatttca gattgctttt gagtttaagt ggtatcaaat 36600

ttcagtatat ttctgtctta tgttaaagaa atatattact aaaacgtaag tgagcaataa 36660

tgtcagctgt caagcactag atttattttt gcaggatatg gagtgcaatg aactgagtca 36720

atatggcgag gtgtatgtga tctgcaggag ttatgccatt taacgtagga aatgtgtggg 36780

actttcccct ctgcactcca gccctcactg taccattagg agacgcaggt ttctttgctg 36840

gagtgtaaaa agttatgtcc atacactcat gtagaatgga tctgaagaaa gcaacattat 36900

atattgtgtt agagaacatt tctatgtgtt taaacttttc actttataga tgaattataa 36960

ctcttaatgc aatgatatag caatttgaaa gcctctccat gttaactgaa ttgaaaatgc 37020

cttgctttaa atatggaata tgatgaaagg gattatctat ttttgcctta gatagctttg 37080

taataatttt tctccaagcg attcaaatct tttgaaaaat gacatgaaat tttcattaaa 37140

aacccttttc ctatgtttgt tgtatacaga aattatgcaa taaaatttct ttataaaaag 37200

gttttcgttt ctgacttctg aaaactgctc tcatttctca tgcagatgca ttggagtttg 37260

ccataggctg ctccgtttac ctccctaccc agctgacatg tctgtagttt ggatagaggg 37320

cattttatct gatggggcat ttctaagctg gggctctgac agtagggtct agtcctactc 37380

tccatacccc agaccccagg agaggcctca tggactttgg caagttgtgt cttgctaact 37440

ctgtaggtga aaaagcagga acgataccat tctgatactt aagcttctgt taaaggaatt 37500

tgacagtaga cttagactgt tgtcaaggat tggtctctag ggaaatatgt taggagttcc 37560

cctttgctca cacgacctca ccttgaaata tctctaacag aagagtttct gtatagtatt 37620

atctgttatg gaaattttag tttaattttt ggttagttcc gtggacttgt ttatttgggg 37680

aggaaaagtg aggtgatgag gaggagtttt gtttttcagc taattctgtg agtcatattc 37740

ctgggatcct aaaattcaca ggccagcaat gtcatggagt gtaccttaga acagtgcttc 37800

tctttaagtg tggtccccag acggcagcat cagcctcacc tgggaacttg ctacaaacac 37860

aagttctggg gccctgtccc agatttactg catcagaagt tccagggtac agccttagca 37920

atatatgttt tagcagtcct ccttgtgatt gccgtgtttg ctcaagttac agaaccactg 37980

ccttggaact gtcagtgaga aaattagcat taattctaaa agcctttcta gctgatgctc 38040

caaaatggct tgttttgtca tcactggttg tctttctatg gagatgggtc caacagtctg 38100

ctctttaatc atgtggatct gtgggtcagc aatgtgtttt ttcaagccaa ttgtctttga 38160

agtggatgcc ttggcaatgg aagcttaaat caggcatttg cattgcaaaa agaaaagcca 38220

actgtcaagg cttgtattac aatccaccct gtgatgctga aacatcatag tcacgagaag 38280

acgacgacat gaatggccta gggcctaggg agcaaagaca ctaaaagtca ggtgtagggc 38340

atgttgtaac ccagaaattg cacatataaa taggattacg gtagcaagac tataataatt 38400

atactggcca tagtccgaaa tcctgtctgt acccattgtc ccaaacttga aaagccaagc 38460

catgcaatag gttagtgatc tcaggggtcc gggaatctgt tcaaagatat ggtaagattg 38520

tggaatattt taacatcttg ggccatctgg tataagtagg tttgaacctc tcctgaattg 38580

gcagattgct ccatatctcc ctgaaccagt ctcttagtcc tgagaacaga tcaactcagt 38640

taaagtgttg tgtcttattt caggaggctg tgacacgggt gggttctcca agttaggtcc 38700

atgtggtaaa agtatttgca tcaggtattt atttatttat tcattatgtt tttctttttg 38760

gctgcgcatg tggaagttcc cgagccaggg ctcgaacgcg ccacggcagc cacacaggct 38820

gctgcagtga caatgctgga tccttaaacc ctgccacaag ggaactcctg catcaggtat 38880

ttaataagag gcaacttcca cagaaacaaa gaaacaaaac acacacatca aggttaatga 38940

ttggagagat tacaactcca cttactgagt tctgatggta gctgatcgag atttcctgac 39000

ggtgggttca aagcatcttt cgagggagta aggcagacag tggcaatctg atagattttc 39060

ctggtttgca gtttgaatgt ctctggtggc tttttgagtg ttctacatag aagtaggcat 39120

gaagattgtc cacataggag tgaggtgact tcttaaagtt gtccacatag gagtgaggtg 39180

acttcttaaa gtttatatca agttgtccca ctttagcttg catggctctg gaaaaaaagg 39240

gcagttttag ttctcaatga ttccaagtca gttaggtgga gaaaaattga aaatgttagt 39300

ttgaagagtt gaagccagat agtggaggaa acaagaattc attatctggt ccagtgtgtt 39360

tccggtagga aacaaaacct gaaaggcaat gaacagaact gcatccaata tccacaagtg 39420

tgcattgtgg tttctattgc aacatgattt ttctctccac aatcatccct atttctatca 39480

acgataactg tagtaagact aacttgtttg caaattaagt ctagttgcaa taaacttggc 39540

ctgattattt acatataagt ccagtaagaa cagtgataga ccataaaagc tcttttaagc 39600

Claims (2)

1.克隆自CNV片段的与猪耳形性状相关的分子标记，其特征在于，所述的分子标记片段为猪五号染色体上Chr 5： 29826401-29866000bp的拷贝数变异，该片段拷贝数为2，3，4，5或6，该分子标记的核苷酸序列如序列表SEQ ID NO： 1所示。

2.权利要求1所述的分子标记在猪耳形标记辅助选择中的应用，当个体拥有该片段的拷贝数>3时，有利于猪拥有非直立的耳形。

Images