实施例1 富集并检测BRCA1/2基因
一、构建样本库
1. DNA的提取
按照常规的组织样本的DNA提取方法提取样本DNA。
2. DNA片段化
按照DNA破碎仪使用说明书,将DNA样本片段化,使片段长度为150-200碱基。
使用Beckman Coulter Ampure Beads试剂盒(货号:A63880)将DNA过柱纯化。
3. DNA样本库质量检测
用生物分析仪进行DNA定性定量分析,确认DNA片段长度峰值合理。
4. DNA末端修补
利用T4聚合酶及Klenow大肠杆菌聚合酶片断,对于cDNA/DNA 5'突出粘末端补平以及3'突出粘末端打平,产生平末端,用于后续的平端连接。反应在PCR扩增仪中进行,20摄氏度,30分钟。
使用Beckman Coulter Ampure Beads试剂盒(货号:A63880)将cDNA/DNA过柱纯化。
5. 在DNA样本3'末端加上碱基A
反应在PCR扩增仪中进行,37℃,30分钟。
使用Beckman Coulter Ampure Beads试剂盒(货号:A63880)将cDNA/DNA过柱纯化。
6. 在DNA两端加上接头
使用Beckman Coulter Ampure Beads试剂盒(货号:A63880)将cDNA/DNA过柱纯化。
7. 扩增6获得的DNA片段样本库
聚合酶链反应(PCR),在PCR扩增仪中进行。
PCR条件:置于PCR扩增仪中,98℃预变性30秒,98℃变性30秒,65℃退火30秒,72℃延伸30秒,共循环15次(cDNA样本库)或者4~6次(DNA样本库)。最后在72℃延伸5分钟。
使用Beckman Coulter Ampure Beads试剂盒(货号:A63880)将PCR扩增产物过柱纯化。
8. 扩增后DNA样本库的质量检测
使用生物分析仪,进行DNA定性定量分析,并确认纯化后片段长度峰值合理,约200bp。
对于得到的DNA样本库,如果DNA浓度小于150纳克/微升,须将样品经过真空浓缩机低温干燥(低于45℃),再用无核酸酶水溶解至所需浓度。
二、针对BRCA1/2基因准备DNA探针库
根据上述的探针设计方法和思路,设计并合成探针进行试验,在5’端有生物素(Biotin)。
三、将DNA样本库与生物素化的DNA探针库杂交
将DNA样本库与杂交缓冲液(10mM Tris-HCl, 2%牛血清白蛋白, pH8.0)混合(混合后,DNA样本库浓度至多不超过50ng/ul),反应条件为95℃ 5分钟,之后保持在65℃。反应在PCR扩增仪中进行。
然后以DNA样本库:探针库为1:100的摩尔比,将探针库加入上述混合物,反应条件为65℃ 5分钟。将杂交反应置于PCR扩增仪中,65℃孵育24小时。
四、得到经杂交富集的BRCA1/2基因片段
1. 准备链霉亲和素磁珠
使用Dynabeads(Life technologies, 货号:11206D)链霉亲和素磁珠或者其它商业化公司链霉亲和素磁珠。将磁珠置于混匀仪上混匀。
磁珠洗涤:混合50微升磁珠和200微升结合缓冲液(10mM Tris-HCl, 2%牛血清白蛋白, pH8.0),在混匀仪上混匀,使用Dynal磁选机或者其它商业化公司磁选机,将磁珠与缓冲液分离纯化,缓冲液弃掉不用。重复三次,每次加入200微升结合缓冲液。
2. 分离杂交产物
混合步骤三中得到的杂交反应混合物与步骤四的1中得到的链霉亲和素磁珠,反复颠倒试管5次。在室温下振摇30分钟。使用Dynal磁选机或者其它商业化公司磁选机,将磁珠分离纯化。
然后向磁珠中加入500微升洗涤缓冲液(磷酸缓冲液,0.1% Tween-20, 0.1% SDS,pH7.4),在65℃孵育10分钟,每隔5分钟混匀一次。使用Dynal磁选机或者其它商业化公司磁选机,将磁珠分离纯化。以上步骤重复三次。
3. DNA富集样本释放
将磁珠与50微升洗脱缓冲液(10mM氢氧化钠溶液)混合,室温孵化10分钟,每隔5分钟混匀一次。使用Dynal磁选机或者其它商业化公司磁选机,将磁珠分离弃掉。此时上清液中即含有富集过的BRCA1/2基因片段DNA样本库。
使用Beckman Coulter Ampure Beads试剂盒(货号:A63880)将样本库过柱纯化。
五、PCR扩增与纯化
将富集cDNA/DNA样本库进一步扩增,为测序仪器上样做准备。
PCR条件:置于PCR扩增仪中,98℃预变性30秒,98℃变性30秒,65℃退火30秒,72℃延伸30秒,共循环15次(cDNA样本库)或者4-6次(DNA样本库)。最后在72℃延伸5分钟。
使用Beckman Coulter Ampure Beads试剂盒(货号:A63880)将PCR扩增产物过柱纯化。
六、采用下一代测序技术检测BRCA1/2基因的基因结构突变
使用TruSeq PE Cluster Kit v3-cBot-HS,使用桥式PCR对DNA样本库模板进行扩增:每个DNA样本片段将会在芯片上形成克隆簇,每条泳道上产生数百万这样的克隆簇。使用Illumina HiSeq2000下一代测序系统,其原理是边合成边测序。和传统Sanger方法相比,利用“可逆性末端终结反应”技术,四种dNTP碱基末端被保护基团封闭,并分别以不同颜色荧光标记。
根据以上方法,采用不同探针进行测试结果如下:
一、不同探针长度检测数据对比
探针长度与探针特异性密切相关,因此我们设计了5种长度的探针,长度分别为80bp、100bp、120bp、140bp、160bp,使用上述探针捕获后进行NGS测序,测序后对数据进行质控,评估捕获序列ontarget率以及目标区域的覆盖率,ontarget率是指位于目标区域的序列在总测序序列中的占比,覆盖率是指目标区域的覆盖情况,具体质控结果见下表:
长度为120bp的探针相比80 bp和100 bp长度探针ontarget率以及目标区域覆盖率均显著提高,进一步加长至140bp以及160bp并未显著提升ontarget率以及目标区域覆盖率,但长探针合成效率显著降低,合成成本较高,因此选用120 bp长度探针。
二、不同GC含量探针对比
探针中GC含量与探针的结合能力高度相关,GC含量过高可能导致大量非特异性结合,因此我们对比分析了未经GC含量优化的探针、优化后GC含量控制在30-65%的探针和优化后GC含量控制在40-55%的探针,捕获后进行NGS检测,质控结果见下表:
GC含量控制在40-55%的探针显著优于GC含量未优化或GC含量30-65%的探针。
三、不同探针overlap长度对比
相邻探针间的相连方式可能与探针覆盖均一性相关,因此我们设计了三种相连形式,分别为10bp gap、5bp overlap和20bp overlap,捕获后进行NGS测序,质控结果如下:
相邻探针间存在5bp overlap相比于10bp gap时50%覆盖率显著升高,与20bp overlap无显著差异。
四、重复区域常规覆盖与优化覆盖方式对比
重复区域在人类基因组中特异性较差,因此我们设计了特殊的覆盖形式,与传统覆盖形式(在重复区域上也均匀地设计了覆盖的探针)相比,重复区域(位置约在BRCA2基因全长329123000-32912400bp左右)捕获后测序结果见图4,上图为采用本专利优化技术设计的探针捕获后的测序结果,下图为采用传统探针捕获后的测序结果,优化后的探针设计对于重复区域覆盖显著优于传统探针设计。在重复区域内存在一个点突变,采用优化后的探针覆盖该位点测序深度为1532X,突变丰度为36%,采用传统探针覆盖该位点测序深度仅为131X,突变丰度为89%,存在较大偏差。
五、外显子临近内含子传统覆盖和优化覆盖方式对比
剪切相关内含子突变在BRCA1/2基因检测中至关重要,因此我们针对外显子临近内含子区域(约在BRCA1基因全长41243000-41243400bp区域范围)进行了特定探针设计,与传统探针设计(不在外显子临近内含子的区域内设计探针)相比,内含子区域捕获结果见图5,上图是传统探针覆盖内含子区域捕获结果,下图为优化后覆盖外显子临近内含子后内含子区域的捕获结果。优化后的探针内含子区域捕获效果显著优于传统探针。
六、检测灵敏度的验证
针对BRCA1的c.4065_4068del突变构建了突变型和野生型的质粒,按照突变型在野生型中拷贝数比例混合而不同丰度的样品,采用上述的探针库进行捕获、测序考察灵敏性,将采用将SEQ ID NO.163-164的探针作为SEQ ID NO.162探针的对照,考察检测灵敏度,每个样本重复测试3次,结果如下:
从表中可以看出,本发明提供的检测探针库和检测方法能够对低丰度的样本也具有较好的检测灵敏度,能够达到0.5%左右的检测灵敏度水平。
采用优化后的探针库对101例乳腺癌和卵巢癌患者白细胞样本和肿瘤组织样本进行BRCA1/2基因突变检测,其中乳腺癌患者76例,卵巢癌患者25例,肿瘤组织与白细胞样本均检出检测结果见下表:其中乳腺癌患者检出突变17例(22%),卵巢癌患者检出突变6例(24%); 外显子区域突变20例,内含子区域突变3例;点突变9例,缺失插入突变13例,融合突变1例。
从上表中可以看出,本发明提供的检测探针库和检测方法能够较好地对BRCA1/2的全外显子区域进行检测。另外,经过DNA提取、PCR扩增和Sanger测序法,以上突变已被验证。
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<210> 31
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<210> 32
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<212> DNA
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cttttgtaag ttcgtcttga tccatctttg tctcccggtt ttaacttacg atacgaatct 60
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<210> 33
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<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 33
cgtaggactt tattttttcg ttcttatact tcttcatcaa gtctgacaat tatgtctaaa 60
gagaggtata gactaaagtc tattgaatct tgtcggatac ccttcatcag tacgtagagt 120
<210> 34
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 34
gagtccaaac aagactctgt ggactactgg acaatctact accactttat ttccttctat 60
gatcaaaacg acttttactg taattccttt caagacgaca aaaatcgttt tcgcaggtct 120
<210> 35
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 35
gtctttcctc tcgaatcgtc ctcaggatcg ggaaagtggg tatgtgtaaa ccgagtccca 60
atggcttctc cccggttctt taatctcagg agtcttctct tgaatagatc actcctactt 120
<210> 36
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 36
acttctcgaa gggacgaagg ttgtgaacaa taaaccattt catttgttat atggaagagt 60
cagatgatcc gtatcgtggc aacgatggct cacagacaga ttcttgtgtc tcctcttaaa 120
<210> 37
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 37
taaataatag taacttctta tcgaatttac tgacgtcatt ggtccattat aaccgtttcc 60
gtagagtcct tgtagtggaa tcactccttt gttttacaag acgatcgaac aaaagaagtg 120
<210> 38
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 38
aaaagaagtg tcacgtcact taaccttctg aactgacgtt tatgtttgtg ggtcctagga 60
aagaactaac caagaaggtt tgtttactcc gtagtcagac tttcggtccc tcaaccagaa 120
<210> 39
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 39
cagactcact gttccttaac caaagtctac tacttctttc tccttgcccg aaccttcttt 60
tattagttct tctcgtttcg tacctaagtt tgaatccata accttggtcc aaaaacacaa 120
<210> 40
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 40
aaaaacaata aattccactt cgtcgtagac ccacactctc actttgttcg cagagacttc 60
tgacgagtcc cgataggaga gtctcactgt aaaattggtg agtccatttt tcgcacacac 120
<210> 41
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 41
taaaagaacc acggtaaata gcaaaaactt cgtctcccta tggtacgttg tattggacta 60
tttcgaggtc gtcctttacc gacttgatct tcgacacaat cttgtcgtac cctcggtcgg 120
<210> 42
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 42
gtcggaagat tgtcgatggg aaggtagtat tcactgagaa gacgggaact cctggacgct 60
ttaggtcttg tttcgtgtag tctttttcca cacataacaa ccggtttgtg actatagaat 120
<210> 43
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 43
caagggaact acagatgtta aagtggaaag aatgtctaag cgtatatgta ccggtttcct 60
tgttgaggta caaaagattt tccggatctc ttgtatatag tccacggaga ggagaaaggg 120
<210> 44
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 44
atttcatcta aacaaaagag taaggtaaat ttcgtcataa ttgaagtgtc ttttcatcac 60
ttatgggata ttcggtctta ggtcttccgg aaagacgact gttcaaactc cacagacgtc 120
<210> 45
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 45
ccggaaagac gactgttcaa actccacaga cgtctatcaa gatggtcatt tttatttctt 60
ggtcctcacc tttccattct ttgtagttac atttctacga caccatagac tgtagaaata 120
<210> 46
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 46
gtaaatagaa agatccagta ggggaagatt tacgggtagt aatctactat ccaccatgta 60
cgtgtcaacg agaccctcag aagtcttatc tttgatgggt agagttctcc tcgagtaatt 120
<210> 47
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 47
gagtaattcc aacaactaca cctcctcgtt gtcgaccttc tcagacccgg tgtgctaaac 60
tgcctttgta gaatgaacgg ttccgttcta gatccattat aaagtagacg acataacctt 120
<210> 48
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 48
atttaatttg aagagggtaa ggaaagtctc ccttggggaa tggaccttag accttagtcg 60
gagaagagac tactgggact tagactagga agacttctgt ctcggggtct cagtcgagca 120
<210> 49
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 49
gcacaaccgt tgtatggtag aagttggaga cgtaactttc aaggggttaa ctttcaacgt 60
cttagacggg tctcaggtcg acgacgagta tgatgactat gacgacccat attacgttac 120
<210> 50
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 50
taccttcttt cacactcgtc cctcttcggt cttaactgtc gaagttgtct ttcccagttg 60
ttttcttaca ggtaccacca cagaccggac tggggtcttc ttaaacactc acataggtat 120
<210> 51
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 51
ctacgagcac atgttcaaac ggtcttttgt ggtgtagtga aattgattag attaatgact 60
tctctgatga gtacaacaat acttttgtcc atatggttct tggaaatgtc ttatggaacg 120
<210> 52
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 52
gtaagacgtc tacgactcaa acacacactt gcctgtgact ttataaaaga tccttaacgc 60
cctcctttta cccatcaatc gataaagaca ttcatattat gataaagagg ggaggaggga 120
<210> 53
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 53
acatacattg gacagaaaag atactagaga aatccccact gggtcagata atttctttct 60
ttttacgact tactccattc atgaactaca atgtttgatt ggtctctata agtaagtcag 120
<210> 54
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 54
aaagaaagtc gtactaaaac ttcagtctcc tctacaccag ttaccttctt tggtggttcc 60
aggtttcgct cgttctctta gggtcctgtc tttccatttc gagggaggga gttcaactgt 120
<210> 55
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 55
gtaaggggac agggagagag aaggagagaa gaaggtctag aagtcccccg atctttagac 60
aacgataccc gggaagtggt tgtacgggtg tccattctcg gaccctcttg gggtctcaag 120
<210> 56
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 56
taaaatctag ttgaccttac ctaccatgtc gacacaccac gaagacacca cttcctcgaa 60
agtagtaagt gggaaccgtg tcattcataa cccacgggac agtctctccc tcctgtgtta 120
<210> 57
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 57
aaacttacga gaaaggaagg acccctaggt cccacaggtg ggttaacacc aacacgtcgg 60
tctacggacc tgtctcctgt taccgaaggt accattccac ggacgtacat ggacacgata 120
<210> 58
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 58
ttaacccgtc tacacactcc gtggacacca ctgggctctc acccacaacc tgtcacatcg 60
tgagatggtc acggtcctcg acctgtggat ggactatggg gtctaggggg tgtcggtgat 120
<210> 59
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 59
tatcgtaggt aaaaatgcct gcattggatc caaagagagg ccaacatttt ttgaaatttt 60
taagacacgc tgcaacaaag caggtattga caaattttat ataactttat aaattacacc 120
<210> 60
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 60
taggaccaat aagtcttaat tggtttgaag aactttcttc agaagctcca ccctataatt 60
ctgaacctgc agaagaatct gaacataaaa acaacaatta cgaaccaaac ctatttaaaa 120
<210> 61
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 61
ccacaaagga aaccatctta taatcagctg gcttcaactc caataatatt caaagagcaa 60
gggctgactc tgccgctgta ccaatctcct gtaaaagaat tagataaatt caaattagac 120
<210> 62
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 62
tttaggaagg aatgttccca atagtagaca taaaagtctt cgcacagtga aaactaaaat 60
ggatcaagca gatgatgttt cctgtccact tctaaattct tgtcttagtg aaaggtatga 120
<210> 63
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 63
acctaaggga tttgctttgt tttattttag tcctgttgtt ctacaatgta cacatgtaac 60
accacaaaga gataagtcag gtatgattaa aaacaatgct ttttattctt agaatactag 120
<210> 64
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 64
aacaattttc cccttttttt acccccagtg gtatgtggga gtttgtttca tacaccaaag 60
tttgtgaagg taaatattct acctggttta tttttatgac ttagtaattg agaatttgac 120
<210> 65
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 65
ggtcgtcaga caccaaaaca tatttctgaa agtctaggag ctgaggtgga tcctgatatg 60
tcttggtcaa gttctttagc tacaccaccc acccttagtt ctactgtgct cataggtaat 120
<210> 66
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 66
acaatacaca taaattttta tcttacagtc agaaatgaag aagcatctga aactgtattt 60
cctcatgata ctactgctgt aagtaaatat gacattgatt agactgttga aattgctaac 120
<210> 67
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 67
gcagaatgtg aaaagctatt tttccaatca tgatgaaagt ctgaagaaaa atgatagatt 60
tatcgcttct gtgacagaca gtgaaaacac aaatcaaaga gaagctgcaa gtcatggtaa 120
<210> 68
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 68
caggatttgg aaaaacatca gggaattcat ttaaagtaaa tagctgcaaa gaccacattg 60
gaaagtcaat gccaaatgtc ctagaagatg aagtatatga aacagttgta gatacctctg 120
<210> 69
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 69
cctctgaaga agatagtttt tcattatgtt tttctaaatg tagaacaaaa aatctacaaa 60
aagtaagaac tagcaagact aggaaaaaaa ttttccatga agcaaacgct gatgaatgtg 120
<210> 70
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 70
gtgaaaaatc taaaaaccaa gtgaaagaaa aatactcatt tgtatctgaa gtggaaccaa 60
atgatactga tccattagat tcaaatgtag caaatcagaa gccctttgag agtggaagtg 120
<210> 71
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 71
agtgacaaaa tctccaagga agttgtaccg tctttggcct gtgaatggtc tcaactaacc 60
ctttcaggtc taaatggagc ccagatggag aaaatacccc tattgcatat ttcttcatgt 120
<210> 72
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 72
atgtgaccaa aatatttcag aaaaagacct attagacaca gagaacaaaa gaaagaaaga 60
ttttcttact tcagagaatt ctttgccacg tatttctagc ctaccaaaat cagagaagcc 120
<210> 73
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 73
agccattaaa tgaggaaaca gtggtaaata agagagatga agagcagcat cttgaatctc 60
atacagactg cattcttgca gtaaagcagg caatatctgg aacttctcca gtggcttctt 120
<210> 74
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 74
tcttcatttc agggtatcaa aaagtctata ttcagaataa gagaatcacc taaagagact 60
ttcaatgcaa gtttttcagg tcatatgact gatccaaact ttaaaaaaga aactgaagcc 120
<210> 75
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 75
agcctctgaa agtggactgg aaatacatac tgtttgctca cagaaggagg actccttatg 60
tccaaattta attgataatg gaagctggcc agccaccacc acacagaatt ctgtagcttt 120
<210> 76
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 76
cagaatctgt agctttgaag aatgcaggtt taatatccac tttgaaaaag aaaacaaata 60
agtttattta tgctatacat gatgaaacat cttataaagg aaaaaaaata ccgaaagacc 120
<210> 77
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 77
gaccaaaaat cagaactaat taactgttca gcccagtttg aagcaaatgc ttttgaagca 60
ccacttacat ttgcaaatgc tgattcaggt acctctgtct tttttttttt gtaaatagta 120
<210> 78
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 78
ggtttattgc attcttctgt gaaaagaagc tgttcacaga atgattctga agaaccaact 60
ttgtccttaa ctagctcttt tgggacaatt ctgaggaaat gttctagaaa tgaaacatgt 120
<210> 79
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 79
atgttctaat aatacagtaa tctctcagga tcttgattat aaagaagcaa aatgtaataa 60
ggaaaaacta cagttattta ttaccccaga agctgattct ctgtcatgcc tgcaggaagg 120
<210> 80
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 80
aaggacagtg tgaaaatgat ccaaaaagca aaaaagtttc agatataaaa gaagaggtct 60
tggctgcagc atgtcaccca gtacaacatt caaaagtgga atacagtgat actgactttc 120
<210> 81
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 81
tttcaatccc agaaaagtct tttatatgat catgaaaatg ccagcactct tattttaact 60
cctacttcca aggatgttct gtcaaaccta gtcatgattt ctagaggcaa agaatcatac 120
<210> 82
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 82
atacaaaatg tcagacaagc tcaaaggtaa caattatgaa tctgatgttg aattaaccaa 60
aaatattccc atggaaaaga atcaagatgt atgtgcttta aatgaaaatt ataaaaacgt 120
<210> 83
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 83
acgttgagct gttgccacct gaaaaataca tgagagtagc atcaccttca agaaaggtac 60
aattcaacca aaacacaaat ctaagagtaa tccaaaaaaa tcaagaagaa actacttcaa 120
<210> 84
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 84
tcaatttcaa aaataactgt caatccagac tctgaagaac ttttctcaga caatgagaat 60
aattttgtct tccaagtagc taatgaaagg aataatcttg ctttaggaaa tactaaggaa 120
<210> 85
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 85
ggaacttcat gaaacagact tgacttgtgt aaacgaaccc attttcaaga actctaccat 60
ggttttatat ggagacacag gtgataaaca agcaacccaa gtgtcaatta aaaaagattt 120
<210> 86
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 86
atttggttta tgttcttgca gaggagaaca aaaatagtgt aaagcagcat ataaaaatga 60
ctctaggtca agatttaaaa tcggacatct ccttgaatat agataaaata ccagaaaaaa 120
<210> 87
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 87
aaaaataatg attacatgaa caaatgggca ggactcttag gtccaatttc aaatcacagt 60
tttggaggta gcttcagaac agcttcaaat aaggaaatca agctctctga acataacatt 120
<210> 88
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 88
cattaagaag agcaaaatgt tcttcaaaga tattgaagaa caatatccta ctagtttagc 60
ttgtgttgaa attgtaaata ccttggcatt agataatcaa aagaaactga gcaagcctca 120
<210> 89
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 89
ctcagtcaat taatactgta tctgcacatt tacagagtag tgtagttgtt tctgattgta 60
aaaatagtca tataacccct cagatgttat tttccaagca ggattttaat tcaaaccata 120
<210> 90
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 90
cataatttaa cacctagcca aaaggcagaa attacagaac tttctactat attagaagaa 60
tcaggaagtc agtttgaatt tactcagttt agaaaaccaa gctacatatt gcagaagagt 120
<210> 91
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 91
gagtacattt gaagtgcctg aaaaccagat gactatctta aagaccactt ctgaggaatg 60
cagagatgct gatcttcatg tcataatgaa tgccccatcg attggtcagg tagacagcag 120
<210> 92
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 92
gcagcaagca atttgaaggt acagttgaaa ttaaacggaa gtttgctggc ctgttgaaaa 60
atgactgtaa caaaagtgct tctggttatt taacagatga aaatgaagtg gggtttaggg 120
<210> 93
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 93
aggggctttt attctgctca tggcacaaaa ctgaatgttt ctactgaagc tctgcaaaaa 60
gctgtgaaac tgtttagtga tattgagaat attagtgagg aaacttctgc agaggtacat 120
<210> 94
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 94
gcagaggtac atccaataag tttatcttca agtaaatgtc atgattctgt tgtttcaatg 60
tttaagatag aaaatcataa tgataaaact gtaagtgaaa aaaataataa atgccaactg 120
<210> 95
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 95
atattacaaa ataatattga aatgactact ggcacttttg ttgaagaaat tactgaaaat 60
tacaagagaa atactgaaaa tgaagataac aaatatactg ctgccagtag aaattctcat 120
<210> 96
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 96
aacttagaat ttgatggcag tgattcaagt aaaaatgata ctgtttgtat tcataaagat 60
gaaacggact tgctatttac tgatcagcac aacatatgtc ttaaattatc tggccagttt 120
<210> 97
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 97
gtttatgaag gagggaaaca ctcagattaa agaagatttg tcagatttaa cttttttgga 60
agttgcgaaa gctcaagaag catgtcatgg taatacttca aataaagaac agttaactgc 120
<210> 98
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 98
ctgctactaa aacggagcaa aatataaaag attttgagac ttctgataca ttttttcaga 60
ctgcaagtgg gaaaaatatt agtgtcgcca aagagtcatt taataaaatt gtaaatttct 120
<210> 99
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 99
ttctttgatc agaaaccaga agaattgcat aacttttcct taaattctga attacattct 60
gacataagaa agaacaaaat ggacattcta agttatgagg aaacagacat agttaaacac 120
<210> 100
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 100
acacaaaata ctgaaagaaa gtgtcccagt tggtactgga aatcaactag tgaccttcca 60
gggacaaccc gaacgtgatg aaaagatcaa agaacctact ctattgggtt ttcatacagc 120
<210> 101
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 101
cagctagcgg gaaaaaagtt aaaattgcaa aggaatcttt ggacaaagtg aaaaaccttt 60
ttgatgaaaa agagcaaggt actagtgaaa tcaccagttt tagccatcaa tgggcaaaga 120
<210> 102
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 102
aagaccctaa agtacagaga ggcctgtaaa gaccttgaat tagcatgtga gaccattgag 60
atcacagctg ccccaaagtg taaagaaatg cagaattctc tcaataatga taaaaacctt 120
<210> 103
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 103
ccttgtttct attgagactg tggtgccacc taagctctta agtgataatt tatgtagaca 60
aactgaaaat ctcaaaacat caaaaagtat ctttttgaaa gttaaagtac atgaaaatgt 120
<210> 104
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 104
atgtagaaaa agaaacagca aaaagtcctg caacttgtta cacaaatcag tccccttatt 60
cagtcattga aaattcagcc ttagcttttt acacaagttg tagtagaaaa acttctgtga 120
<210> 105
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 105
gtgagtcaga cttcattact tgaagcaaaa aaatggctta gagaaggaat atttgatggt 60
caaccagaaa gaataaatac tgcagattat gtaggaaatt atttgtatga aaataattca 120
<210> 106
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 106
ttcaaacagt actatagctg aaaatgacaa aaatcatctc tccgaaaaac aagatactta 60
tttaagtaac agtagcatgt ctaacagcta ttcctaccat tctgatgagg tatataatga 120
<210> 107
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 107
atgattcagg atatctctca aaaaataaac ttgattctgg tattgagcca gtattgaaga 60
atgttgaaga tcaaaaaaac actagttttt ccaaagtaat atccaatgta aaagatgcaa 120
<210> 108
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 108
gcaaatgcat acccacaaac tgtaaatgaa gatatttgcg ttgaggaact tgtgactagc 60
tcttcaccct gcaaaaataa aaatgcagcc attaaattgt ccatatctaa tagtaataat 120
<210> 109
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 109
taattttgag gtagggccac ctgcatttag gatagccagt ggtaaaatcg tttgtgtttc 60
acatgaaaca attaaaaaag tgaaagacat atttacagac agtttcagta aagtaattaa 120
<210> 110
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 110
ttaaggaaaa caacgagaat aaatcaaaaa tttgccaaac gaaaattatg gcaggttgtt 60
acgaggcatt ggatgattca gaggatattc ttcataactc tctagataat gatgaatgta 120
<210> 111
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 111
tgtagcacgc attcacataa ggtttttgct gacattcaga gtgaagaaat tttacaacat 60
aaccaaaata tgtctggatt ggagaaagtt tctaaaatat caccttgtga tgttagtttg 120
<210> 112
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 112
tttggaaact tcagatatat gtaaatgtag tatagggaag cttcataagt cagtctcatc 60
tgcaaatact tgtgggattt ttagcacagc aagtggaaaa tctgtccagg tatcagatgc 120
<210> 113
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 113
atgcttcatt acaaaacgca agacaagtgt tttctgaaat agaagatagt accaagcaag 60
tcttttccaa agtattgttt aaaagtaacg aacattcaga ccagctcaca agagaagaaa 120
<210> 114
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 114
gaaaatactg ctatacgtac tccagaacat ttaatatccc aaaaaggctt ttcatataat 60
gtggtaaatt catctgcttt ctctggattt agtacagcaa gtggaaagca agtttccatt 120
<210> 115
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 115
cattttagaa agttccttac acaaagttaa gggagtgtta gaggaatttg atttaatcag 60
aactgagcat agtcttcact attcacctac gtctagacaa aatgtatcaa aaatacttcc 120
<210> 116
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 116
ttcctcgtgt tgataagaga aacccagagc actgtgtaaa ctcagaaatg gaaaaaacct 60
gcagtaaaga atttaaatta tcaaataact taaatgttga aggtggttct tcagaaaata 120
<210> 117
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 117
aataatcact ctattaaagt ttctccatat ctctctcaat ttcaacaaga caaacaacag 60
ttggtattag gaaccaaagt gtcacttgtt gagaacattc atgttttggg aaaagaacag 120
<210> 118
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 118
acaggcttca cctaaaaacg taaaaatgga aattggtaaa actgaaactt tttctgatgt 60
tcctgtgaaa acaaatatag aagtttgttc tacttactcc aaagattcag aaaactactt 120
<210> 119
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 119
actttgaaac agaagcagta gaaattgcta aagcttttat ggaagatgat gaactgacag 60
attctaaact gccaagtcat gccacacatt ctctttttac atgtcccgaa aatgaggaaa 120
<210> 120
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 120
gaaatggttt tgtcaaattc aagaattgga aaaagaagag gagagcccct tatcttagtg 60
ggtaagtgtt catttttacc tttcgtgttg ccaatcacta tttttaaagt gtttattcag 120
<210> 121
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 121
ttctttttag gagaaccctc aatcaaaaga aacttattaa atgaatttga caggataata 60
gaaaatcaag aaaaatcctt aaaggcttca aaaagcactc cagatggtaa aattagcttt 120
<210> 122
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 122
tgtttcctag gcacaataaa agatcgaaga ttgtttatgc atcatgtttc tttagagccg 60
attacctgtg taccctttcg gtaagacatg tttaaatttt tctaaattct aatacagtat 120
<210> 123
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 123
ttctccccat tgcagcacaa ctaaggaacg tcaagagata cagaatccaa attttaccgc 60
acctggtcaa gaatttctgt ctaaatctca tttgtatgaa catctgactt tggaaaaatc 120
<210> 124
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 124
aatcttcaag caatttagca gtttcaggac atccatttta tcaagtttct gctacaagaa 60
atgaaaaaat gagacacttg attactacag gcagaccaac caaagtcttt gttccacctt 120
<210> 125
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 125
accttttaaa actaaatcac attttcacag agttgaacag tgtgttagga atattaactt 60
ggaggaaaac agacaaaagc aaaacattga tggacatggc tctgatgata gtaaaaataa 120
<210> 126
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 126
aaataagatt aatgacaatg agattcatca gtttaacaaa aacaactcca atcaagcagt 60
agctgtaact ttcacaaagt gtgaagaaga acctttaggt attgtatgac aatttgtgtg 120
<210> 127
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 127
agtatttatt ctttgataga tttaattaca agtcttcaga atgccagaga tatacaggat 60
atgcgaatta agaagaaaca aaggcaacgc gtctttccac agccaggcag tctgtatctt 120
<210> 128
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 128
ctgtatcttg caaaaacatc cactctgcct cgaatctctc tgaaagcagc agtaggaggc 60
caagttccct ctgcgtgttc tcataaacag gtatgtgttt gtctacaata ctgatggctt 120
<210> 129
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 129
ttttttgtgt gtgtttattt tgtgtagctg tatacgtatg gcgtttctaa acattgcata 60
aaaattaaca gcaaaaatgc agagtctttt cagtttcaca ctgaagatta ttttggtaag 120
<210> 130
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 130
tttggtaagg aaagtttatg gactggaaaa ggaatacagt tggctgatgg tggatggctc 60
ataccctcca atgatggaaa ggctggaaaa gaagaatttt ataggtactc tatgcaaaaa 120
<210> 131
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 131
acttttattt gttcagggct ctgtgtgaca ctccaggtgt ggatccaaag cttatttcta 60
gaatttgggt ttataatcac tatagatgga tcatatggaa actggcagct atggaatgtg 120
<210> 132
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 132
tggaatgtgc ctttcctaag gaatttgcta atagatgcct aagcccagaa agggtgcttc 60
ttcaactaaa atacaggcaa gtttaaagca ttacattacg taatcatata cggcagtatg 120
<210> 133
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 133
tcacttttag atatgatacg gaaattgata gaagcagaag atcggctata aaaaagataa 60
tggaaaggga tgacacagct gcaaaaacac ttgttctctg tgtttctgac ataatttcat 120
<210> 134
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 134
ttcattgagc gcaaatatat ctgaaacttc tagcaataaa actagtagtg cagataccca 60
aaaagtggcc attattgaac ttacagatgg gtggtatgct gttaaggccc agttagatcc 120
<210> 135
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 135
gatcctcccc tcttagctgt cttaaagaat ggcagactga cagttggtca gaagattatt 60
cttcatggag cagaactggt gggctctcct gatgcctgta cacctcttga agccccagaa 120
<210> 136
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 136
cagaatctct tatgttaaag gtaaattaat ttgcactctt ggtaaaaatc agtcattgat 60
tcagttaaat tctagaagtt ttacatttaa attttaaatg cttactaagg atgctcaatt 120
<210> 137
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 137
tcaatatatt tattaatttg tccagatttc tgctaacagt actcggcctg ctcgctggta 60
taccaaactt ggattctttc ctgaccctag accttttcct ctgcccttat catcgctttt 120
<210> 138
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 138
atcgcttttc agtgatggag gaaatgttgg ttgtgttgat gtaattattc aaagagcata 60
ccctatacag gtatgatgta ttcttgaaac ttaccatata tttctttctt ttgatacaat 120
<210> 139
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 139
gtaacacatt attacagtgg atggagaaga catcatctgg attatacata tttcgcaatg 60
aaagagagga agaaaaggaa gcagcaaaat atgtggaggc ccaacaaaag agactagaag 120
<210> 140
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 140
gactagaagc cttattcact aaaattcagg aggaatttga agaacatgaa ggtaaaatta 60
gttatatggt acacattgtt atttctaata tgagaacaaa gtcttagaga ctttgaattt 120
<210> 141
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 141
tagtgaatta ataatccttt tgttttctta gaaaacacaa caaaaccata tttaccatca 60
cgtgcactaa caagacagca agttcgtgct ttgcaagatg gtgcagagct ttatgaagca 120
<210> 142
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 142
tatgaagcag tgaagaatgc agcagaccca gcttaccttg aggtgagaga gtaagaggac 60
atataatgag gcttgatgat tattcaaggt gagaagctgt tttagactct ctggccatca 120
<210> 143
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 143
atcttaaatg gtcacagggt tatttcagtg aagagcagtt aagagccttg aataatcaca 60
ggcaaatgtt gaatgataag aaacaagctc agatccagtt ggaaattagg aaggccatgg 120
<210> 144
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 144
aggccatgga atctgctgaa caaaaggaac aaggtttatc aagggatgtc acaaccgtgt 60
ggaagttgcg tattgtaagc tattcaaaaa aagaaaaaga ttcaggtaag tatgtaaatg 120
<210> 145
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 145
tctccaaaca gttatactga gtatttggcg tccatcatca gatttatatt ctctgttaac 60
agaaggaaag agatacagaa tttatcatct tgcaacttca aaatctaaaa gtaaatctga 120
<210> 146
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 146
taaatctgaa agagctaaca tacagttagc agcgacaaaa aaaactcagt atcaacaact 60
accggtacaa acctttcatt gtaatttttc agttttgata agtgcttgtt agtttatgga 120
<210> 147
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 147
atctccatat gttgaatttt tgttttgttt tctgtaggtt tcagatgaaa ttttatttca 60
gatttaccag ccacgggagc cccttcactt cagcaaattt ttagatccag actttcagcc 120
<210> 148
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 148
ctttcagcca tcttgttctg aggtggacct aataggattt gtcgtttctg ttgtgaaaaa 60
aacaggtaat gcacaatata gttaattttt tttattgatt cttttaaaaa acattgtctt 120
<210> 149
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 149
cttttctttt ttttccattc taggacttgc ccctttcgtc tatttgtcag acgaatgtta 60
caatttactg gcaataaagt tttggataga ccttaatgag gacattatta agcctcatat 120
<210> 150
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 150
gttaattgct gcaagcaacc tccagtggcg accagaatcc aaatcaggcc ttcttacttt 60
atttgctgga gatttttctg tgttttctgc tagtccaaaa gagggccact ttcaagagac 120
<210> 151
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 151
ttcaagagac attcaacaaa atgaaaaata ctgttgaggt aaggttactt ttcagcatca 60
ccacacattt tggtattttt ctattttgac agtccagtat caaggaaata gcttttatac 120
<210> 152
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 152
tatacaaatt ggatagttga ggtagtatgt gaggtaaagt ttaatcatat attaattgcc 60
atgggggaat ggggaaatga catgaacctc aggagcagca actagaaaga gaagaatgac 120
<210> 153
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 153
cacttatttt cttagaatat tgacatactt tgcaatgaag cagaaaacaa gcttatgcat 60
atactgcatg caaatgatcc caagtggtcc accccaacta aagactgtac ttcagggccg 120
<210> 154
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 154
tcagggccgt acactgctca aatcattcct ggtacaggaa acaagcttct ggtaagttaa 60
tgtaaactca aggaatatta taagaagtat atatggaggc catcgtatat tctgttgtat 120
<210> 155
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 155
ctacgttttc atttttttat cagatgtctt ctcctaattg tgagatatat tatcaaagtc 60
ctttatcact ttgtatggcc aaaaggaagt ctgtttccac acctgtctca gcccagatga 120
<210> 156
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 156
cttcaaagtc ttgtaaaggg gagaaagaga ttgatgacca aaagaactgc aaaaagagaa 60
gagccttgga tttcttgagt agactgcctt tacctccacc tgttagtccc atttgtacat 120
<210> 157
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 157
ttgtttctcc ggctgcacag aaggcatttc agccaccaag gagttgtggc accaaatacg 60
aaacacccat aaagaaaaaa gaactgaatt ctcctcagat gactccattt aaaaaattca 120
<210> 158
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 158
atgaaatttc tcttttggaa agtaattcaa tagctgacga agaacttgca ttgataaata 60
cccaagctct tttgtctggt tcaacaggag aaaaacaatt tatatctgtc agtgaatcca 120
<210> 159
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 159
ctaggactgc tcccaccagt tcagaagatt atctcagact gaaacgacgt tgtactacat 60
ctctgatcaa agaacaggag agttcccagg ccagtacgga agaatgtgag aaaaataagc 120
<210> 160
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 160
aggacacaat tacaactaaa aaatatatct aagcatttgc aaaggcgaca ataaattatt 60
gacgcttaac ctttccagtt tataagactg gaatataatt tcaaaccaca cattagtact 120
<210> 161
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 161
tatgttgcac aatgagaaaa gaaattagtt tcaaatttac ctcagcgttt gtgtatcggg 60
caaaaatcgt tttgcccgat tccgtattgg tatacttttg cttcagttgc atatcttaaa 120
<210> 162
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 162
cacccaggat cctttcttga ttggttcttc caaacaaatg aggcatcagt ctgaaagcca 60
gggagttggt ctgagtgaca aggaattggt ttcagatgat gaagaaagag gaacgggctt 120
<210> 163
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 163
ttgattggtt cttccaaaca aatgaggcat cagtctgaaa gccagggagt tggtctgagt 60
gacaaggaat tggtttcaga tgatgaagaa agaggaacgg gcttggaaga aaataatcaa 120
<210> 164
<211> 120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 164
aggagaattt attatcattg aagaatagct taaatgactg cagtaaccag gtaatattgg 60
caaaggcatc tcaggaacat caccttagtg aggaaacaaa atgttctgct agcttgtttt 120