CN111394385A - 一种快速鉴定水稻双向启动子的方法 - Google Patents

一种快速鉴定水稻双向启动子的方法 Download PDF

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CN111394385A
CN111394385A CN202010234073.2A CN202010234073A CN111394385A CN 111394385 A CN111394385 A CN 111394385A CN 202010234073 A CN202010234073 A CN 202010234073A CN 111394385 A CN111394385 A CN 111394385A
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刘鹏
李鑫琪
张韬
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Yangzhou University
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Abstract

本发明公开了一种快速鉴定水稻双向启动子的方法，该方法将双向启动子OsBDP1和OsBDP2两端分别连接两个不同的荧光蛋白GFP和RFP，在原生质体转化后观察两侧的荧光信号来检测其活性。该鉴定方法一方面利用瞬时转化缩短了实验时间，另一方面通过同时检测两侧荧光蛋白活性而减少了工作量，为快速鉴定水稻中的双向启动子提供了一个高效途径。

Description

一种快速鉴定水稻双向启动子的方法
技术领域
本发明涉及生物技术，特别涉及一种快速鉴定水稻双向启动子的方法。
背景技术
双向启动子是指位于一对“头对头”基因的两个TSSs之间，富含高GC且长度一般不超过1.0Kb，可分别驱动正义链与反义链下游基因表达的DNA序列，如图1所示。在基因工程领域，利用双向启动子在两个方向上控制基因转录这一特性，在其两端连接两个不同的目的基因。这样不仅降低载体构建难度，而且减少转录调控元件的引入，避免了可能因调控元件之间同源序列的存在所导致的转基因“共抑制”现象。因此双向启动子可用于多基因转化，即启动两个及两个以上基因的同时表达，避免了因重复利用启动子而导致的基因沉默等结果。目前双向启动子的鉴定主要有生物信息学预测和实验验证两种方法。生物信息学的鉴定方法本质上都是通过不同的软件预测启动子中存在的 motif结合位点、转录调控区域、DHS区域或者核小体缺失区域，筛选符合条件的双向启动子进行表达。实验鉴定的方法主要基于启动子驱动报告基因表达的方式来检测，主要分为稳定转化和瞬时表达两个途径。当前研究最常使用的方法是将双向启动子两边分别连接GFP和GUS，通过稳定转化在转基因植株中进行定性和定量分析。但在原生质体瞬时转化体系中，对双向启动子是否能同时启动两侧基因的表达，这一功能的验证的研究尚未报道。
发明内容
发明目的：本发明目的是提供一种快速鉴定水稻双向启动子的方法。
技术方案：本发明提供一种快速鉴定水稻双向启动子的方法，包括如下步骤：
(1)选取已通过水稻稳定转化验证的两个双向启动子，OsBDP1和OsBDP2；
启动子序列参见：OsBDP1：
Functional characterization of open chromatin in bidirectionalpromoters of rice；
OsBDP2：
Bidirectional Promoter-Based CRISPR-Cas9 Systems for Plant GenomeEditing。
所在染色体位置：
BDP1:Chr2:19649523-19649914；BDP2:Chr2:25453280-25455516。
(2)构建双报告载体GFP-RFPreporter(SEQ ID NO.1)；(3)将OsBDP1和OsBDP2 扩增后分别插入双报告载体GFP-RFP reporter(SEQ ID NO.1)，构建OsBDP1 reporter和OsBDP2 reporter载体(SEQ ID NO.2，SEQ ID NO.3)；(4)水稻原生质体的制备与转化；(5)转化后对OsBDP1、OsBDP2的活性进行验证。
进一步地，所述步骤(2)中双报告载体GFP-RFP reporter构建方法为：以pCambia1301载体为模板，扩增Ubiquitin启动子；以载体pGWB454为模板，扩增RFP 编码序列；使用限制性内切酶NcoI和EcoRI将p1300LV载体线性化，用无缝连接试剂盒将上述3个片段快速连接，转化至大肠杆菌DH5α中，筛选阳性转化子。构建负对照载体。所述步骤(3)扩增的引物序列分别为：SEQ ID NO.6、SEQ ID NO.7、
SEQ ID NO.8、SEQ ID NO.9。
OsBDP1-F:5’-gacgtcctcggaggaggccatggtttttcactttgtcagtg-3’(SEQ IDNO.6)；
OsBDP1-R:5’-tctcttcttcttaggagccatgcagggctcacttactcgca-3’(SEQ IDNO.7)；
OsBDP2-F:5’-gacgtcctcggaggaggccatagctgcaaacataacaaatatac-3’(SEQ IDNO.8)；
OsBDP2-R:5’-tctcttcttcttaggagccatctcgtctcctc tctactgttg-3’(SEQ IDNO.9)。
所述负对照载体构建方法为：使用限制性内切酶KpnI和BamHI将GFP-RFPreporter载体线性化，将2个mini35s或一段无功能序列插入线性化载体，连接后转化至大肠杆菌DH5α中，筛选阳性转化子，得到2个负对照载体mini35s reporter(SEQ ID NO.4)、promoterless reporter(SEQ ID NO.5)。
有益效果：本发明将双向启动子两端分别连接两个不同的荧光蛋白，GFP和RFP，在原生质体转化后观察两侧的荧光信号来检测其活性。一方面瞬时转化缩短了实验时间，另一方面通过同时检测两侧荧光蛋白活性而减少了工作量。该方法操作简单，耗时短，能够在水稻全基因组水平上对双向启动子活性进行快速筛选鉴定，为研究水稻中的调控元件提供了实验支持。为快速鉴定水稻中的双向启动子提供了一个有效途径。
附图说明
图1是双向启动子的结构示意图；
图2是正负载体结构示意图及共聚焦显微镜观察荧光蛋白表达结果图，GFP-RFPreporter载体是正对照载体，载体结构为CaMV35S启动GFP表达，Ubiquitin启动RFP 表达；mini35s reporter和promoterless reporter是负对照，mini35s reporter载体结构为mini35s启动GFP表达，mini35s启动RFP表达；promoterless reporter载体是由一段无启动子活性的序列同时启动GFP和RFP表达；
图3OsBDP1和OsBDP2启动荧光蛋白表达结果图。
具体实施方式
本实施例鉴定方法如下：
1.正对照载体：以pCambia1301载体为模板，扩增Ubiquitin启动子；以载体pGWB454 为模板，扩增RFP基因；使用限制性内切酶NcoI和EcoRI将p1300LV载体线性化，用无缝连接试剂盒将上述3个片段快速连接，转化至大肠杆菌DH5α中，筛选阳性转化子。得到正对照载体GFP-RFP reporter(如图2所示)。
2.负对照载体：使用限制性内切酶KpnI和BamHI将GFP-RFP reporter载体线性化，将2个mini35s或一段无功能序列插入线性化载体，连接后转化至大肠杆菌DH5α中，筛选阳性转化子。得到2个负对照载体mini35s reporter和promoterless reporter(如图1 所示)。
3.候选双向启动子的选取：
选取已通过稳定转化验证的两个双向启动子OsBDP1、OsBDP2来验证该体系的可行性。OsBDP1调控的一对基因是LOC_Os02g33070和LOC_Os02g33080，OsBDP2调控的一对基因是LOC_Os02g42314和LOC_Os02g42320。
4.OsBDP1、OsBDP2的分离克隆及表达载体构建
利用PCR法，以水稻日本晴基因组DNA为模板，通过特异引物扩增(引物序列如SEQID NO.6、SEQ ID NO.7、SEQ ID NO.8、SEQ ID NO.9)。PCR扩增得到的目的片段OsBDP1/OsBDP2分别与BamHI和KpnI双酶切后的双报告载体GFP-RFP reporter 连接，构建成OsBDP1reporter和OsBDP2 reporter载体。
5.水稻原生质体制备与转化
5.1水稻原生质体制备：(1)叶片准备。取无菌培养14天左右生长良好的新鲜水稻植株40-60株，去掉根部，备用。(2)酶解。将水稻叶片和茎段用锋利刀片切成宽度为0.5mm的条状，速度要快，防止叶片脱水，将切好的叶片放入盛有10mL酶解液的培养皿中，置于密闭容器中用真空泵抽真空30min，用封口膜封口，置于转速为60rpm 的摇床上25℃黑暗条件下过夜酶解。(3)过滤。将过夜酶解的粗酶液通过预先用2mL W5 washing Buffer润洗过的孔径40μm的细胞筛进行过滤，去除酶解不充分的杂质。(4) 提纯。将过滤过的细胞酶解液转移到两个15mL离心管中，每管6mL细胞酶解液，用装有长针头的注射器吸取16mL 0.55M蔗糖，每管细胞酶解液底部加入8mL 0.55M蔗糖，注意：加蔗糖时，针头伸入底部，推动注射器，随着液面升高，缓慢上移注射器，动作要轻柔，防止对原生质体细胞造成破坏。1000g离心5min。(5)清洗。取50mL离心管一个，加入10mL Washing buffer，置于试管架上。用装有长针头的注射器吸取8mL Washing buffer，再将针头伸到上步离心后的中间细胞层，慢慢吸取该层的所有原生质体，然后小心转移至盛有10mL Washing buffer的50mL离心管中。轻弹混匀，100g室温离心5min。(6)重复清洗一次。移除上清，沿壁慢慢加入10mL Washing buffer，轻弹混匀，100g离心2min。(7)重悬。去掉上清，加入5mL Washing buffer，注意沿壁缓慢加入，轻弹混匀，使细胞保持悬浮。(8)计数。吸取100μL细胞用于计数，取6-7μL细胞置于细胞计数板进行计数，如果细胞数目太多可采用MGG稀释后再进行计数。(9)细胞稀释。根据计数结果取适量细胞悬浮液进行离心，然后用MGG Buffer重悬细胞，最终将细胞稀释至1×106个/mL，用于原生质体转化。
5.2水稻原生质体的转化：(1)质粒的准备。采用质粒小提试剂盒(TIANGEN， Cat#DP-106)提取以下质粒：GFP-RFP reporter，mini35s reporter,promoterless reporter,OsBDP1 reporter和OsBDP2 reporter,确保质粒浓度尽量在500ng/μL以上，整个过程最好为无菌操作。质粒置于-20℃冰箱冻存。使用前以离心机最高转速进行离心10min处理，使杂质沉淀于离心管底部，避免因杂质影响原生质体转化的效率。处理好的质粒用 MGG Buffer稀释到实验所需质粒浓度，稀释后充分混匀备用。(2)PEG介导的转化。每管质粒中加入200μL按照2.2.6制备过程制备的原生质体细胞，轻弹混匀。然后，每管加入210-230μL40％PEG(PEG加入量与质粒体积相关，确保等体积加入)，轻弹混匀，直至细胞均匀悬浮于PEG当中，从第一管加入PEG开始计时，计时20min。从第一管加入到最后一管加入的时间尽量控制在5min以内。(3)清洗。加入PEG反应20min 后，每管加入1mLW5 Washing Buffer，终止反应，轻轻颠倒混匀，250g离心5min。(4) 重复清洗一次。用枪轻轻吸掉上清，重新加入800μLWashing Buffer，轻轻颠倒混匀， 150g离心5min，轻轻吸掉上清。(5)细胞重悬与培养。取40mm培养皿加入2mLW5 Buffer，再取培养皿中的W5 Buffer重悬离心管中的细胞，并全部转移至培养皿中，25℃黑暗培养，两天后在显微镜下观察荧光。
6.激光共聚焦显微镜观察
使用Zeiss 700激光共聚集显微镜进行观察。GFP、RFP使用的激发波长分别为：488nm，543nm，吸收波长分别为490-540nm，415-735nm。所有的荧光实验至少重复三次。观察结果如图2、图3所示，转化后OsBDP1、OsBDP2两个转化子中均能观察到明显的GFP和RFP荧光信号。
序列表
<110> 扬州大学
<120> 一种快速鉴定水稻双向启动子的方法
<160> 5
<170> SIPOSequenceListing 1.0
<210> 1
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<212> DNA
<213> 人工序列(Artificial Sequence)
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agcttggcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa 60
cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc 120
accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgctagag cagcttgagc 180
ttggatcaga ttgtcgtttc ccgccttcag tttaaactat cagtgtttga caggatatat 240
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gaaaaggttt atccgttcgt ccatttgtat gtgcatgcca accacagggt tcccctcggg 360
atcaaagtac tttgatccaa cccctccgct gctatagtgc agtcggcttc tgacgttcag 420
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cgcgccctgc aactcgccgg ggccgatgtt ctgttagtcg attccgatcc ccagggcagt 1680
gcccgcgatt gggcggccgt gcgggaagat caaccgctaa ccgttgtcgg catcgaccgc 1740
ccgacgattg accgcgacgt gaaggccatc ggccggcgcg acttcgtagt gatcgacgga 1800
gcgccccagg cggcggactt ggctgtgtcc gcgatcaagg cagccgactt cgtgctgatt 1860
ccggtgcagc caagccctta cgacatatgg gccaccgccg acctggtgga gctggttaag 1920
cagcgcattg aggtcacgga tggaaggcta caagcggcct ttgtcgtgtc gcgggcgatc 1980
aaaggcacgc gcatcggcgg tgaggttgcc gaggcgctgg ccgggtacga gctgcccatt 2040
cttgagtccc gtatcacgca gcgcgtgagc tacccaggca ctgccgccgc cggcacaacc 2100
gttcttgaat cagaacccga gggcgacgct gcccgcgagg tccaggcgct ggccgctgaa 2160
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ggaggcggca tggaaaatca agaacaacca ggcaccgacg ccgtggaatg ccccatgtgt 2520
ggaggaacgg gcggttggcc aggcgtaagc ggctgggttg tctgccggcc ctgcaatggc 2580
actggaaccc ccaagcccga ggaatcggcg tgacggtcgc aaaccatccg gcccggtaca 2640
aatcggcgcg gcgctgggtg atgacctggt ggagaagttg aaggccgcgc aggccgccca 2700
gcggcaacgc atcgaggcag aagcacgccc cggtgaatcg tggcaagcgg ccgctgatcg 2760
aatccgcaaa gaatcccggc aaccgccggc agccggtgcg ccgtcgatta ggaagccgcc 2820
caagggcgac gagcaaccag attttttcgt tccgatgctc tatgacgtgg gcacccgcga 2880
tagtcgcagc atcatggacg tggccgtttt ccgtctgtcg aagcgtgacc gacgagctgg 2940
cgaggtgatc cgctacgagc ttccagacgg gcacgtagag gtttccgcag ggccggccgg 3000
catggccagt gtgtgggatt acgacctggt actgatggcg gtttcccatc taaccgaatc 3060
catgaaccga taccgggaag ggaagggaga caagcccggc cgcgtgttcc gtccacacgt 3120
tgcggacgta ctcaagttct gccggcgagc cgatggcgga aagcagaaag acgacctggt 3180
agaaacctgc attcggttaa acaccacgca cgttgccatg cagcgtacga agaaggccaa 3240
gaacggccgc ctggtgacgg tatccgaggg tgaagccttg attagccgct acaagatcgt 3300
aaagagcgaa accgggcggc cggagtacat cgagatcgag ctagctgatt ggatgtaccg 3360
cgagatcaca gaaggcaaga acccggacgt gctgacggtt caccccgatt actttttgat 3420
cgatcccggc atcggccgtt ttctctaccg cctggcacgc cgcgccgcag gcaaggcaga 3480
agccagatgg ttgttcaaga cgatctacga acgcagtggc agcgccggag agttcaagaa 3540
gttctgtttc accgtgcgca agctgatcgg gtcaaatgac ctgccggagt acgatttgaa 3600
ggaggaggcg gggcaggctg gcccgatcct agtcatgcgc taccgcaacc tgatcgaggg 3660
cgaagcatcc gccggttcct aatgtacgga gcagatgcta gggcaaattg ccctagcagg 3720
ggaaaaaggt cgaaaaggtc tctttcctgt ggatagcacg tacattggga acccaaagcc 3780
gtacattggg aaccggaacc cgtacattgg gaacccaaag ccgtacattg ggaaccggtc 3840
acacatgtaa gtgactgata taaaagagaa aaaaggcgat ttttccgcct aaaactcttt 3900
aaaacttatt aaaactctta aaacccgcct ggcctgtgca taactgtctg gccagcgcac 3960
agccgaagag ctgcaaaaag cgcctaccct tcggtcgctg cgctccctac gccccgccgc 4020
ttcgcgtcgg cctatcgcgg ccgctggccg ctcaaaaatg gctggcctac ggccaggcaa 4080
tctaccaggg cgcggacaag ccgcgccgtc gccactcgac cgccggcgcc cacatcaagg 4140
caccctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg 4200
agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt 4260
cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag 4320
tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg 4380
gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct cttccgcttc 4440
ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc 4500
aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc 4560
aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag 4620
gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc 4680
gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt 4740
tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct 4800
ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg 4860
ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct 4920
tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat 4980
tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg 5040
ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa 5100
aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt 5160
ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc 5220
tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgcattc 5280
taggtactaa aacaattcat ccagtaaaat ataatatttt attttctccc aatcaggctt 5340
gatccccagt aagtcaaaaa atagctcgac atactgttct tccccgatat cctccctgat 5400
cgaccggacg cagaaggcaa tgtcatacca cttgtccgcc ctgccgcttc tcccaagatc 5460
aataaagcca cttactttgc catctttcac aaagatgttg ctgtctccca ggtcgccgtg 5520
ggaaaagaca agttcctctt cgggcttttc cgtctttaaa aaatcataca gctcgcgcgg 5580
atctttaaat ggagtgtctt cttcccagtt ttcgcaatcc acatcggcca gatcgttatt 5640
cagtaagtaa tccaattcgg ctaagcggct gtctaagcta ttcgtatagg gacaatccga 5700
tatgtcgatg gagtgaaaga gcctgatgca ctccgcatac agctcgataa tcttttcagg 5760
gctttgttca tcttcatact cttccgagca aaggacgcca tcggcctcac tcatgagcag 5820
attgctccag ccatcatgcc gttcaaagtg caggaccttt ggaacaggca gctttccttc 5880
cagccatagc atcatgtcct tttcccgttc cacatcatag gtggtccctt tataccggct 5940
gtccgtcatt tttaaatata ggttttcatt ttctcccacc agcttatata ccttagcagg 6000
agacattcct tccgtatctt ttacgcagcg gtatttttcg atcagttttt tcaattccgg 6060
tgatattctc attttagcca tttattattt ccttcctctt ttctacagta tttaaagata 6120
ccccaagaag ctaattataa caagacgaac tccaattcac tgttccttgc attctaaaac 6180
cttaaatacc agaaaacagc tttttcaaag ttgttttcaa agttggcgta taacatagta 6240
tcgacggagc cgattttgaa accgcggtga tcacaggcag caacgctctg tcatcgttac 6300
aatcaacatg ctaccctccg cgagatcatc cgtgtttcaa acccggcagc ttagttgccg 6360
ttcttccgaa tagcatcggt aacatgagca aagtctgccg ccttacaacg gctctcccgc 6420
tgacgccgtc ccggactgat gggctgcctg tatcgagtgg tgattttgtg ccgagctgcc 6480
ggtcggggag ctgttggctg gctggtggca ggatatattg tggtgtaaac aaattgacgc 6540
ttagacaact taataacaca ttgcggacgt ttttaatgta ctgaattaac gccgaattaa 6600
ttcgggggat ctggatttta gtactggatt ttggttttag gaattagaaa ttttattgat 6660
agaagtattt tacaaataca aatacatact aagggtttct tatatgctca acacatgagc 6720
gaaaccctat aggaacccta attcccttat ctgggaacta ctcacacatt attatggaga 6780
aactcgagct tgtcgatcga cagatccggt cggcatctac tctatttctt tgccctcgga 6840
cgagtgctgg ggcgtcggtt tccactatcg gcgagtactt ctacacagcc atcggtccag 6900
acggccgcgc ttctgcgggc gatttgtgta cgcccgacag tcccggctcc ggatcggacg 6960
attgcgtcgc atcgaccctg cgcccaagct gcatcatcga aattgccgtc aaccaagctc 7020
tgatagagtt ggtcaagacc aatgcggagc atatacgccc ggagtcgtgg cgatcctgca 7080
agctccggat gcctccgctc gaagtagcgc gtctgctgct ccatacaagc caaccacggc 7140
ctccagaaga agatgttggc gacctcgtat tgggaatccc cgaacatcgc ctcgctccag 7200
tcaatgaccg ctgttatgcg gccattgtcc gtcaggacat tgttggagcc gaaatccgcg 7260
tgcacgaggt gccggacttc ggggcagtcc tcggcccaaa gcatcagctc atcgagagcc 7320
tgcgcgacgg acgcactgac ggtgtcgtcc atcacagttt gccagtgata cacatgggga 7380
tcagcaatcg cgcatatgaa atcacgccat gtagtgtatt gaccgattcc ttgcggtccg 7440
aatgggccga acccgctcgt ctggctaaga tcggccgcag cgatcgcatc catagcctcc 7500
gcgaccggtt gtagaacagc gggcagttcg gtttcaggca ggtcttgcaa cgtgacaccc 7560
tgtgcacggc gggagatgca ataggtcagg ctctcgctaa actccccaat gtcaagcact 7620
tccggaatcg ggagcgcggc cgatgcaaag tgccgataaa cataacgatc tttgtagaaa 7680
ccatcggcgc agctatttac ccgcaggaca tatccacgcc ctcctacatc gaagctgaaa 7740
gcacgagatt cttcgccctc cgagagctgc atcaggtcgg agacgctgtc gaacttttcg 7800
atcagaaact tctcgacaga cgtcgcggtg agttcaggct ttttcatatc tcattgcccc 7860
ccgggatctg cgaaagctcg agagagatag atttgtagag agagactggt gatttcagcg 7920
tgtcctctcc aaatgaaatg aacttcctta tatagaggaa ggtcttgcga aggatagtgg 7980
gattgtgcgt catcccttac gtcagtggag atatcacatc aatccacttg ctttgaagac 8040
gtggttggaa cgtcttcttt ttccacgatg ctcctcgtgg gtgggggtcc atctttggga 8100
ccactgtcgg cagaggcatc ttgaacgata gcctttcctt tatcgcaatg atggcatttg 8160
taggtgccac cttccttttc tactgtcctt ttgatgaagt gacagatagc tgggcaatgg 8220
aatccgagga ggtttcccga tattaccctt tgttgaaaag tctcaatagc cctttggtct 8280
tctgagactg tatctttgat attcttggag tagacgagag tgtcgtgctc caccatgtta 8340
tcacatcaat ccacttgctt tgaagacgtg gttggaacgt cttctttttc cacgatgctc 8400
ctcgtgggtg ggggtccatc tttgggacca ctgtcggcag aggcatcttg aacgatagcc 8460
tttcctttat cgcaatgatg gcatttgtag gtgccacctt ccttttctac tgtccttttg 8520
atgaagtgac agatagctgg gcaatggaat ccgaggaggt ttcccgatat taccctttgt 8580
tgaaaagtct caatagccct ttggtcttct gagactgtat ctttgatatt cttggagtag 8640
acgagagtgt cgtgctccac catgttggca agctgctcta gccaatacgc aaaccgcctc 8700
tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag 8760
cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt 8820
tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca 8880
caggaaacag ctatgaccat gattacgaat tcccgatcta gtaacataga tgacaccgcg 8940
cgcgataatt tatcctagtt tgcgcgctat attttgtttt ctatcgcgta ttaaatgtat 9000
aattgcggga ctctaatcat aaaaacccat ctcataaata acgtcatgca ttacatgtta 9060
attattacat gcttaacgta attcaacaga aattatatga taatcatcgc aagaccggca 9120
acaggattca atcttaagaa actttattgc caaatgtttg aacgatcggg gaaattcgag 9180
ctctaagctt aggcgccggt ggagtggcgg ccctcggcgc gctcgtactg ttccacgatg 9240
gtgtagtcct cgttgtggga ggtgatgtcc agcttgatgt cggtcttgta ggcgccgggc 9300
agctgcacgg gcttcttggc catgtaggtg gtcttgacct cggcgtcgta gtggccgccg 9360
tccttcagct tcagcctcat cttgatctcg cccttcaggg cgccgtcctc ggggtacatc 9420
cgctcggtgg aggcctccca gcccatggtc ttcttctgca ttacggggcc gtcggagggg 9480
aagttggtgc cgcgcagctt caccttgtag atgaactcgc cgtcctgcag ggaggagtcc 9540
tgggtcacgg tcaccacgcc gccgtcctcg aagttcatca cgcgctccca cttgaagccc 9600
tcggggaagg acagcttcaa gtagtcgggg atgtcggcgg ggtgcttcac gtaggccttg 9660
gagccgtact ggaactgagg ggacaggatg tcccaggcga agggcagggg gccgcccttg 9720
gtcaccttca gcttggcggt ctgggtgccc tcgtaggggc ggccctcgcc ctcgccctcg 9780
atctcgaact cgtggccgtt cacggagccc tccatgcgca ccttgaagcg catgaactcc 9840
ttgatgacgt cctcggagga ggccattgtt ataacctttc tcttcttctt aggagccatg 9900
gatccctgca gaagtaacac caaacaacag ggtgagcatc gacaaaagaa acagtaccaa 9960
gcaaataaat agcgtatgaa ggcagggcta aaaaaatcca catatagctg ctgcatatgc 10020
catcatccaa gtatatcaag atcaaaataa ttataaaaca tacttgttta ttataataga 10080
taggtactca aggttagagc atatgaatag atgctgcata tgccatcatg tatatgcatc 10140
agtaaaaccc acatcaacat gtatacctat cctagatcga tatttccatc catcttaaac 10200
tcgtaactat gaagatgtat gacacacaca tacagttcca aaattaataa atacaccagg 10260
tagtttgaaa cagtattcta ctccgatcta gaacgaatga acgaccgccc aaccacacca 10320
catcatcaca accaagcgaa caaaaagcat ctctgtatat gcatcagtaa aacccgcatc 10380
aacatgtata cctatcctag atcgatattt ccatccatca tcttcaattc gtaactatga 10440
atatgtatgg cacacacata cagatccaaa attaataaat ccaccaggta gtttgaaaca 10500
gaattctact ccgatctaga acgaccgccc aaccagacca catcatcaca accaagacaa 10560
aaaaaagcat gaaaagatga cccgacaaac aagtgcacgg catatattga aataaaggaa 10620
aagggcaaac caaaccctat gcaacgaaac aaaaaaaatc atgaaatcga tcccgtctgc 10680
ggaacggcta gagccatccc aggattcccc aaagagaaac actggcaagt tagcaatcag 10740
aacgtgtctg acgtacaggt cgcatccgtg tacgaacgct agcagcacgg atctaacaca 10800
aacacggatc taacacaaac atgaacagaa gtagaactac cgggccctaa ccatggaccg 10860
gaacgccgat ctagagaagg tagagagggg ggggggggga ggacgagcgg cgtaccttga 10920
agcggaggtg ccgacgggtg gatttggggg agatctggtt gtgtgtgtgt gcgctccgaa 10980
caacacgagg ttggggaaag agggtgtgga gggggtgtct atttattacg gcgggcgagg 11040
aagggaaagc gaaggagcgg tgggaaagga atcccccgta gctgccggtg ccgtgagagg 11100
aggaggaggc cgcctgccgt gccggctcac gtctgccgct ccgccacgca atttctggat 11160
gccgacagcg gagcaagtcc aacggtggag cggaactctc gagaggggtc cagaggcagc 11220
gacagagatg ccgtgccgtc tgcttcgctt ggcccgacgc gacgctgctg gttcgctggt 11280
tggtgtccgt tagactcgtc gacggcgttt aacaggctgg cattatctac tcgaaacaag 11340
aaaaatgttt ccttagtttt tttaatttct taaagggtat ttgtttaatt tttagtcact 11400
ttattttatt ctattttata tctaaattat taaataaaaa aactaaaata gagttttagt 11460
tttcttaatt tagaggctaa aatagaataa aatagatgta ctaaaaaaat tagtctataa 11520
aaaccattaa ccctaaaccc taaatggatg tactaataaa atggatgaag tattatatag 11580
gtgaagctat ttgcaaaaaa aaaggagaac acatgcacac taaaaagata aaactgtaga 11640
gtcctgttgt caaaatactc aattgtcctt tagaccatgt ctaactgttc atttatatga 11700
ttctctaaaa cactgatatt attgtagtac tatagattat attattcgta gagtaaagtt 11760
taaatatatg tataaagata gataaactgc acttcaaaca agtgtgacaa aaaaaatatg 11820
tggtaatttt ttataactta gacatgcaat gctcattatc tctagagagg ggcacgaccg 11880
ggtcacgctg cactgcagat ggagtcaaag attcaaatag aggacctaac agaactcgcc 11940
gtaaagactg gcgaacagtt catacagagt ctcttacgac tcaatgacaa gaagaaaatc 12000
ttcgtcaaca tggtggagca cgacacactt gtctactcca aaaatatcaa agatacagtc 12060
tcagaagacc aaagggcaat tgagactttt caacaaaggg taatatccgg aaacctcctc 12120
ggattccatt gcccagctat ctgtcacttt attgtgaaga tagtggaaaa ggaaggtggc 12180
tcctacaaat gccatcattg cgataaagga aaggccatcg ttgaagatgc ctctgccgac 12240
agtggtccca aagatggacc cccacccacg aggagcatcg tggaaaaaga agacgttcca 12300
accacgtctt caaagcaagt ggattgatgt gatatctcca ctgacgtaag ggatgacgca 12360
caatcccact atccttcgca agacccttcc tctatataag gaagttcatt tcatttggag 12420
agaacaggta ccatggctcc taagaagaag agaaaggtta taacaatggt gagcaagggc 12480
gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc 12540
cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg 12600
aagctgatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg 12660
ggctacggcc tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc 12720
aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc 12780
aactacaaga cacgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag 12840
ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac 12900
tacaacagcc acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac 12960
ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag 13020
aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag 13080
tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg 13140
accgccgccg ggatcactct cggcatggac gaggcagcta gatccacctg agtcgagtcg 13200
acggatatat gaagatgaag atgaaatatt tggtgtgtca aataaaaagc ttgtgtgctt 13260
aagtttgtgt ttttttcttg gcttgttgtg ttatgaattt gtggcttttt ctaatattaa 13320
atgaatgtaa gatcacatta taatgaataa acaaatgttt ctataatcca ttgtgaatgt 13380
tttgttggat ctcttctgca gcatataact actgtatgtg ctatggtatg gactatggaa 13440
tatgattaaa gataagg 13457
<210> 2
<211> 11283
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 2
agcttggcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa 60
cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc 120
accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgctagag cagcttgagc 180
ttggatcaga ttgtcgtttc ccgccttcag tttaaactat cagtgtttga caggatatat 240
tggcgggtaa acctaagaga aaagagcgtt tattagaata acggatattt aaaagggcgt 300
gaaaaggttt atccgttcgt ccatttgtat gtgcatgcca accacagggt tcccctcggg 360
atcaaagtac tttgatccaa cccctccgct gctatagtgc agtcggcttc tgacgttcag 420
tgcagccgtc ttctgaaaac gacatgtcgc acaagtccta agttacgcga caggctgccg 480
ccctgccctt ttcctggcgt tttcttgtcg cgtgttttag tcgcataaag tagaatactt 540
gcgactagaa ccggagacat tacgccatga acaagagcgc cgccgctggc ctgctgggct 600
atgcccgcgt cagcaccgac gaccaggact tgaccaacca acgggccgaa ctgcacgcgg 660
ccggctgcac caagctgttt tccgagaaga tcaccggcac caggcgcgac cgcccggagc 720
tggccaggat gcttgaccac ctacgccctg gcgacgttgt gacagtgacc aggctagacc 780
gcctggcccg cagcacccgc gacctactgg acattgccga gcgcatccag gaggccggcg 840
cgggcctgcg tagcctggca gagccgtggg ccgacaccac cacgccggcc ggccgcatgg 900
tgttgaccgt gttcgccggc attgccgagt tcgagcgttc cctaatcatc gaccgcaccc 960
ggagcgggcg cgaggccgcc aaggcccgag gcgtgaagtt tggcccccgc cctaccctca 1020
ccccggcaca gatcgcgcac gcccgcgagc tgatcgacca ggaaggccgc accgtgaaag 1080
aggcggctgc actgcttggc gtgcatcgct cgaccctgta ccgcgcactt gagcgcagcg 1140
aggaagtgac gcccaccgag gccaggcggc gcggtgcctt ccgtgaggac gcattgaccg 1200
aggccgacgc cctggcggcc gccgagaatg aacgccaaga ggaacaagca tgaaaccgca 1260
ccaggacggc caggacgaac cgtttttcat taccgaagag atcgaggcgg agatgatcgc 1320
ggccgggtac gtgttcgagc cgcccgcgca cgtctcaacc gtgcggctgc atgaaatcct 1380
ggccggtttg tctgatgcca agctggcggc ctggccggcc agcttggccg ctgaagaaac 1440
cgagcgccgc cgtctaaaaa ggtgatgtgt atttgagtaa aacagcttgc gtcatgcggt 1500
cgctgcgtat atgatgcgat gagtaaataa acaaatacgc aaggggaacg catgaaggtt 1560
atcgctgtac ttaaccagaa aggcgggtca ggcaagacga ccatcgcaac ccatctagcc 1620
cgcgccctgc aactcgccgg ggccgatgtt ctgttagtcg attccgatcc ccagggcagt 1680
gcccgcgatt gggcggccgt gcgggaagat caaccgctaa ccgttgtcgg catcgaccgc 1740
ccgacgattg accgcgacgt gaaggccatc ggccggcgcg acttcgtagt gatcgacgga 1800
gcgccccagg cggcggactt ggctgtgtcc gcgatcaagg cagccgactt cgtgctgatt 1860
ccggtgcagc caagccctta cgacatatgg gccaccgccg acctggtgga gctggttaag 1920
cagcgcattg aggtcacgga tggaaggcta caagcggcct ttgtcgtgtc gcgggcgatc 1980
aaaggcacgc gcatcggcgg tgaggttgcc gaggcgctgg ccgggtacga gctgcccatt 2040
cttgagtccc gtatcacgca gcgcgtgagc tacccaggca ctgccgccgc cggcacaacc 2100
gttcttgaat cagaacccga gggcgacgct gcccgcgagg tccaggcgct ggccgctgaa 2160
attaaatcaa aactcatttg agttaatgag gtaaagagaa aatgagcaaa agcacaaaca 2220
cgctaagtgc cggccgtccg agcgcacgca gcagcaaggc tgcaacgttg gccagcctgg 2280
cagacacgcc agccatgaag cgggtcaact ttcagttgcc ggcggaggat cacaccaagc 2340
tgaagatgta cgcggtacgc caaggcaaga ccattaccga gctgctatct gaatacatcg 2400
cgcagctacc agagtaaatg agcaaatgaa taaatgagta gatgaatttt agcggctaaa 2460
ggaggcggca tggaaaatca agaacaacca ggcaccgacg ccgtggaatg ccccatgtgt 2520
ggaggaacgg gcggttggcc aggcgtaagc ggctgggttg tctgccggcc ctgcaatggc 2580
actggaaccc ccaagcccga ggaatcggcg tgacggtcgc aaaccatccg gcccggtaca 2640
aatcggcgcg gcgctgggtg atgacctggt ggagaagttg aaggccgcgc aggccgccca 2700
gcggcaacgc atcgaggcag aagcacgccc cggtgaatcg tggcaagcgg ccgctgatcg 2760
aatccgcaaa gaatcccggc aaccgccggc agccggtgcg ccgtcgatta ggaagccgcc 2820
caagggcgac gagcaaccag attttttcgt tccgatgctc tatgacgtgg gcacccgcga 2880
tagtcgcagc atcatggacg tggccgtttt ccgtctgtcg aagcgtgacc gacgagctgg 2940
cgaggtgatc cgctacgagc ttccagacgg gcacgtagag gtttccgcag ggccggccgg 3000
catggccagt gtgtgggatt acgacctggt actgatggcg gtttcccatc taaccgaatc 3060
catgaaccga taccgggaag ggaagggaga caagcccggc cgcgtgttcc gtccacacgt 3120
tgcggacgta ctcaagttct gccggcgagc cgatggcgga aagcagaaag acgacctggt 3180
agaaacctgc attcggttaa acaccacgca cgttgccatg cagcgtacga agaaggccaa 3240
gaacggccgc ctggtgacgg tatccgaggg tgaagccttg attagccgct acaagatcgt 3300
aaagagcgaa accgggcggc cggagtacat cgagatcgag ctagctgatt ggatgtaccg 3360
cgagatcaca gaaggcaaga acccggacgt gctgacggtt caccccgatt actttttgat 3420
cgatcccggc atcggccgtt ttctctaccg cctggcacgc cgcgccgcag gcaaggcaga 3480
agccagatgg ttgttcaaga cgatctacga acgcagtggc agcgccggag agttcaagaa 3540
gttctgtttc accgtgcgca agctgatcgg gtcaaatgac ctgccggagt acgatttgaa 3600
ggaggaggcg gggcaggctg gcccgatcct agtcatgcgc taccgcaacc tgatcgaggg 3660
cgaagcatcc gccggttcct aatgtacgga gcagatgcta gggcaaattg ccctagcagg 3720
ggaaaaaggt cgaaaaggtc tctttcctgt ggatagcacg tacattggga acccaaagcc 3780
gtacattggg aaccggaacc cgtacattgg gaacccaaag ccgtacattg ggaaccggtc 3840
acacatgtaa gtgactgata taaaagagaa aaaaggcgat ttttccgcct aaaactcttt 3900
aaaacttatt aaaactctta aaacccgcct ggcctgtgca taactgtctg gccagcgcac 3960
agccgaagag ctgcaaaaag cgcctaccct tcggtcgctg cgctccctac gccccgccgc 4020
ttcgcgtcgg cctatcgcgg ccgctggccg ctcaaaaatg gctggcctac ggccaggcaa 4080
tctaccaggg cgcggacaag ccgcgccgtc gccactcgac cgccggcgcc cacatcaagg 4140
caccctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg 4200
agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt 4260
cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag 4320
tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg 4380
gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct cttccgcttc 4440
ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc 4500
aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc 4560
aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag 4620
gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc 4680
gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt 4740
tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct 4800
ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg 4860
ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct 4920
tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat 4980
tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg 5040
ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa 5100
aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt 5160
ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc 5220
tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgcattc 5280
taggtactaa aacaattcat ccagtaaaat ataatatttt attttctccc aatcaggctt 5340
gatccccagt aagtcaaaaa atagctcgac atactgttct tccccgatat cctccctgat 5400
cgaccggacg cagaaggcaa tgtcatacca cttgtccgcc ctgccgcttc tcccaagatc 5460
aataaagcca cttactttgc catctttcac aaagatgttg ctgtctccca ggtcgccgtg 5520
ggaaaagaca agttcctctt cgggcttttc cgtctttaaa aaatcataca gctcgcgcgg 5580
atctttaaat ggagtgtctt cttcccagtt ttcgcaatcc acatcggcca gatcgttatt 5640
cagtaagtaa tccaattcgg ctaagcggct gtctaagcta ttcgtatagg gacaatccga 5700
tatgtcgatg gagtgaaaga gcctgatgca ctccgcatac agctcgataa tcttttcagg 5760
gctttgttca tcttcatact cttccgagca aaggacgcca tcggcctcac tcatgagcag 5820
attgctccag ccatcatgcc gttcaaagtg caggaccttt ggaacaggca gctttccttc 5880
cagccatagc atcatgtcct tttcccgttc cacatcatag gtggtccctt tataccggct 5940
gtccgtcatt tttaaatata ggttttcatt ttctcccacc agcttatata ccttagcagg 6000
agacattcct tccgtatctt ttacgcagcg gtatttttcg atcagttttt tcaattccgg 6060
tgatattctc attttagcca tttattattt ccttcctctt ttctacagta tttaaagata 6120
ccccaagaag ctaattataa caagacgaac tccaattcac tgttccttgc attctaaaac 6180
cttaaatacc agaaaacagc tttttcaaag ttgttttcaa agttggcgta taacatagta 6240
tcgacggagc cgattttgaa accgcggtga tcacaggcag caacgctctg tcatcgttac 6300
aatcaacatg ctaccctccg cgagatcatc cgtgtttcaa acccggcagc ttagttgccg 6360
ttcttccgaa tagcatcggt aacatgagca aagtctgccg ccttacaacg gctctcccgc 6420
tgacgccgtc ccggactgat gggctgcctg tatcgagtgg tgattttgtg ccgagctgcc 6480
ggtcggggag ctgttggctg gctggtggca ggatatattg tggtgtaaac aaattgacgc 6540
ttagacaact taataacaca ttgcggacgt ttttaatgta ctgaattaac gccgaattaa 6600
ttcgggggat ctggatttta gtactggatt ttggttttag gaattagaaa ttttattgat 6660
agaagtattt tacaaataca aatacatact aagggtttct tatatgctca acacatgagc 6720
gaaaccctat aggaacccta attcccttat ctgggaacta ctcacacatt attatggaga 6780
aactcgagct tgtcgatcga cagatccggt cggcatctac tctatttctt tgccctcgga 6840
cgagtgctgg ggcgtcggtt tccactatcg gcgagtactt ctacacagcc atcggtccag 6900
acggccgcgc ttctgcgggc gatttgtgta cgcccgacag tcccggctcc ggatcggacg 6960
attgcgtcgc atcgaccctg cgcccaagct gcatcatcga aattgccgtc aaccaagctc 7020
tgatagagtt ggtcaagacc aatgcggagc atatacgccc ggagtcgtgg cgatcctgca 7080
agctccggat gcctccgctc gaagtagcgc gtctgctgct ccatacaagc caaccacggc 7140
ctccagaaga agatgttggc gacctcgtat tgggaatccc cgaacatcgc ctcgctccag 7200
tcaatgaccg ctgttatgcg gccattgtcc gtcaggacat tgttggagcc gaaatccgcg 7260
tgcacgaggt gccggacttc ggggcagtcc tcggcccaaa gcatcagctc atcgagagcc 7320
tgcgcgacgg acgcactgac ggtgtcgtcc atcacagttt gccagtgata cacatgggga 7380
tcagcaatcg cgcatatgaa atcacgccat gtagtgtatt gaccgattcc ttgcggtccg 7440
aatgggccga acccgctcgt ctggctaaga tcggccgcag cgatcgcatc catagcctcc 7500
gcgaccggtt gtagaacagc gggcagttcg gtttcaggca ggtcttgcaa cgtgacaccc 7560
tgtgcacggc gggagatgca ataggtcagg ctctcgctaa actccccaat gtcaagcact 7620
tccggaatcg ggagcgcggc cgatgcaaag tgccgataaa cataacgatc tttgtagaaa 7680
ccatcggcgc agctatttac ccgcaggaca tatccacgcc ctcctacatc gaagctgaaa 7740
gcacgagatt cttcgccctc cgagagctgc atcaggtcgg agacgctgtc gaacttttcg 7800
atcagaaact tctcgacaga cgtcgcggtg agttcaggct ttttcatatc tcattgcccc 7860
ccgggatctg cgaaagctcg agagagatag atttgtagag agagactggt gatttcagcg 7920
tgtcctctcc aaatgaaatg aacttcctta tatagaggaa ggtcttgcga aggatagtgg 7980
gattgtgcgt catcccttac gtcagtggag atatcacatc aatccacttg ctttgaagac 8040
gtggttggaa cgtcttcttt ttccacgatg ctcctcgtgg gtgggggtcc atctttggga 8100
ccactgtcgg cagaggcatc ttgaacgata gcctttcctt tatcgcaatg atggcatttg 8160
taggtgccac cttccttttc tactgtcctt ttgatgaagt gacagatagc tgggcaatgg 8220
aatccgagga ggtttcccga tattaccctt tgttgaaaag tctcaatagc cctttggtct 8280
tctgagactg tatctttgat attcttggag tagacgagag tgtcgtgctc caccatgtta 8340
tcacatcaat ccacttgctt tgaagacgtg gttggaacgt cttctttttc cacgatgctc 8400
ctcgtgggtg ggggtccatc tttgggacca ctgtcggcag aggcatcttg aacgatagcc 8460
tttcctttat cgcaatgatg gcatttgtag gtgccacctt ccttttctac tgtccttttg 8520
atgaagtgac agatagctgg gcaatggaat ccgaggaggt ttcccgatat taccctttgt 8580
tgaaaagtct caatagccct ttggtcttct gagactgtat ctttgatatt cttggagtag 8640
acgagagtgt cgtgctccac catgttggca agctgctcta gccaatacgc aaaccgcctc 8700
tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag 8760
cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt 8820
tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca 8880
caggaaacag ctatgaccat gattacgaat tcccgatcta gtaacataga tgacaccgcg 8940
cgcgataatt tatcctagtt tgcgcgctat attttgtttt ctatcgcgta ttaaatgtat 9000
aattgcggga ctctaatcat aaaaacccat ctcataaata acgtcatgca ttacatgtta 9060
attattacat gcttaacgta attcaacaga aattatatga taatcatcgc aagaccggca 9120
acaggattca atcttaagaa actttattgc caaatgtttg aacgatcggg gaaattcgag 9180
ctctaagctt aggcgccggt ggagtggcgg ccctcggcgc gctcgtactg ttccacgatg 9240
gtgtagtcct cgttgtggga ggtgatgtcc agcttgatgt cggtcttgta ggcgccgggc 9300
agctgcacgg gcttcttggc catgtaggtg gtcttgacct cggcgtcgta gtggccgccg 9360
tccttcagct tcagcctcat cttgatctcg cccttcaggg cgccgtcctc ggggtacatc 9420
cgctcggtgg aggcctccca gcccatggtc ttcttctgca ttacggggcc gtcggagggg 9480
aagttggtgc cgcgcagctt caccttgtag atgaactcgc cgtcctgcag ggaggagtcc 9540
tgggtcacgg tcaccacgcc gccgtcctcg aagttcatca cgcgctccca cttgaagccc 9600
tcggggaagg acagcttcaa gtagtcgggg atgtcggcgg ggtgcttcac gtaggccttg 9660
gagccgtact ggaactgagg ggacaggatg tcccaggcga agggcagggg gccgcccttg 9720
gtcaccttca gcttggcggt ctgggtgccc tcgtaggggc ggccctcgcc ctcgccctcg 9780
atctcgaact cgtggccgtt cacggagccc tccatgcgca ccttgaagcg catgaactcc 9840
ttgatgacgt cctcggagga ggccatggtt tttcactttg tcagtgtccg tcggatgctc 9900
tcatcctgac cattttgcgg cgcgaaaccg aagataagga tgggaatatg ggattgagat 9960
gagaacgccg gaactaaccg gataaaagaa agaaagccaa gtggtttgga aagcaactcg 10020
agtagtctgg gcctaaacca tcctagtttg ggcctcccgg ctcccggccc ggccaaaccc 10080
gatccatcgg aagcacgcga ggcgaagtga ttcgtgcagc gggttccacc aaccgcgcgg 10140
cggatcggac ggtggtgcgc gcctctcagc cgccgagtcc acggcgacca ccgcctcacc 10200
aacccacgca accaatgcca tgtgaataac agaataagtg cgagtaagtg agccctgcat 10260
ggctcctaag aagaagagaa aggttataac aatggtgagc aagggcgagg agctgttcac 10320
cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt 10380
gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagc tgatctgcac 10440
caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgggct acggcctgca 10500
gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc 10560
cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacacg 10620
cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga 10680
cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa 10740
cgtctatatc accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca 10800
caacatcgag gacggcggcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg 10860
cgacggcccc gtgctgctgc ccgacaacca ctacctgagc taccagtccg ccctgagcaa 10920
agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat 10980
cactctcggc atggacgagg cagctagatc cacctgagtc gagtcgacgg atatatgaag 11040
atgaagatga aatatttggt gtgtcaaata aaaagcttgt gtgcttaagt ttgtgttttt 11100
ttcttggctt gttgtgttat gaatttgtgg ctttttctaa tattaaatga atgtaagatc 11160
acattataat gaataaacaa atgtttctat aatccattgt gaatgttttg ttggatctct 11220
tctgcagcat ataactactg tatgtgctat ggtatggact atggaatatg attaaagata 11280
agg 11283
<210> 3
<211> 13128
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 3
agcttggcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa 60
cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc 120
accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgctagag cagcttgagc 180
ttggatcaga ttgtcgtttc ccgccttcag tttaaactat cagtgtttga caggatatat 240
tggcgggtaa acctaagaga aaagagcgtt tattagaata acggatattt aaaagggcgt 300
gaaaaggttt atccgttcgt ccatttgtat gtgcatgcca accacagggt tcccctcggg 360
atcaaagtac tttgatccaa cccctccgct gctatagtgc agtcggcttc tgacgttcag 420
tgcagccgtc ttctgaaaac gacatgtcgc acaagtccta agttacgcga caggctgccg 480
ccctgccctt ttcctggcgt tttcttgtcg cgtgttttag tcgcataaag tagaatactt 540
gcgactagaa ccggagacat tacgccatga acaagagcgc cgccgctggc ctgctgggct 600
atgcccgcgt cagcaccgac gaccaggact tgaccaacca acgggccgaa ctgcacgcgg 660
ccggctgcac caagctgttt tccgagaaga tcaccggcac caggcgcgac cgcccggagc 720
tggccaggat gcttgaccac ctacgccctg gcgacgttgt gacagtgacc aggctagacc 780
gcctggcccg cagcacccgc gacctactgg acattgccga gcgcatccag gaggccggcg 840
cgggcctgcg tagcctggca gagccgtggg ccgacaccac cacgccggcc ggccgcatgg 900
tgttgaccgt gttcgccggc attgccgagt tcgagcgttc cctaatcatc gaccgcaccc 960
ggagcgggcg cgaggccgcc aaggcccgag gcgtgaagtt tggcccccgc cctaccctca 1020
ccccggcaca gatcgcgcac gcccgcgagc tgatcgacca ggaaggccgc accgtgaaag 1080
aggcggctgc actgcttggc gtgcatcgct cgaccctgta ccgcgcactt gagcgcagcg 1140
aggaagtgac gcccaccgag gccaggcggc gcggtgcctt ccgtgaggac gcattgaccg 1200
aggccgacgc cctggcggcc gccgagaatg aacgccaaga ggaacaagca tgaaaccgca 1260
ccaggacggc caggacgaac cgtttttcat taccgaagag atcgaggcgg agatgatcgc 1320
ggccgggtac gtgttcgagc cgcccgcgca cgtctcaacc gtgcggctgc atgaaatcct 1380
ggccggtttg tctgatgcca agctggcggc ctggccggcc agcttggccg ctgaagaaac 1440
cgagcgccgc cgtctaaaaa ggtgatgtgt atttgagtaa aacagcttgc gtcatgcggt 1500
cgctgcgtat atgatgcgat gagtaaataa acaaatacgc aaggggaacg catgaaggtt 1560
atcgctgtac ttaaccagaa aggcgggtca ggcaagacga ccatcgcaac ccatctagcc 1620
cgcgccctgc aactcgccgg ggccgatgtt ctgttagtcg attccgatcc ccagggcagt 1680
gcccgcgatt gggcggccgt gcgggaagat caaccgctaa ccgttgtcgg catcgaccgc 1740
ccgacgattg accgcgacgt gaaggccatc ggccggcgcg acttcgtagt gatcgacgga 1800
gcgccccagg cggcggactt ggctgtgtcc gcgatcaagg cagccgactt cgtgctgatt 1860
ccggtgcagc caagccctta cgacatatgg gccaccgccg acctggtgga gctggttaag 1920
cagcgcattg aggtcacgga tggaaggcta caagcggcct ttgtcgtgtc gcgggcgatc 1980
aaaggcacgc gcatcggcgg tgaggttgcc gaggcgctgg ccgggtacga gctgcccatt 2040
cttgagtccc gtatcacgca gcgcgtgagc tacccaggca ctgccgccgc cggcacaacc 2100
gttcttgaat cagaacccga gggcgacgct gcccgcgagg tccaggcgct ggccgctgaa 2160
attaaatcaa aactcatttg agttaatgag gtaaagagaa aatgagcaaa agcacaaaca 2220
cgctaagtgc cggccgtccg agcgcacgca gcagcaaggc tgcaacgttg gccagcctgg 2280
cagacacgcc agccatgaag cgggtcaact ttcagttgcc ggcggaggat cacaccaagc 2340
tgaagatgta cgcggtacgc caaggcaaga ccattaccga gctgctatct gaatacatcg 2400
cgcagctacc agagtaaatg agcaaatgaa taaatgagta gatgaatttt agcggctaaa 2460
ggaggcggca tggaaaatca agaacaacca ggcaccgacg ccgtggaatg ccccatgtgt 2520
ggaggaacgg gcggttggcc aggcgtaagc ggctgggttg tctgccggcc ctgcaatggc 2580
actggaaccc ccaagcccga ggaatcggcg tgacggtcgc aaaccatccg gcccggtaca 2640
aatcggcgcg gcgctgggtg atgacctggt ggagaagttg aaggccgcgc aggccgccca 2700
gcggcaacgc atcgaggcag aagcacgccc cggtgaatcg tggcaagcgg ccgctgatcg 2760
aatccgcaaa gaatcccggc aaccgccggc agccggtgcg ccgtcgatta ggaagccgcc 2820
caagggcgac gagcaaccag attttttcgt tccgatgctc tatgacgtgg gcacccgcga 2880
tagtcgcagc atcatggacg tggccgtttt ccgtctgtcg aagcgtgacc gacgagctgg 2940
cgaggtgatc cgctacgagc ttccagacgg gcacgtagag gtttccgcag ggccggccgg 3000
catggccagt gtgtgggatt acgacctggt actgatggcg gtttcccatc taaccgaatc 3060
catgaaccga taccgggaag ggaagggaga caagcccggc cgcgtgttcc gtccacacgt 3120
tgcggacgta ctcaagttct gccggcgagc cgatggcgga aagcagaaag acgacctggt 3180
agaaacctgc attcggttaa acaccacgca cgttgccatg cagcgtacga agaaggccaa 3240
gaacggccgc ctggtgacgg tatccgaggg tgaagccttg attagccgct acaagatcgt 3300
aaagagcgaa accgggcggc cggagtacat cgagatcgag ctagctgatt ggatgtaccg 3360
cgagatcaca gaaggcaaga acccggacgt gctgacggtt caccccgatt actttttgat 3420
cgatcccggc atcggccgtt ttctctaccg cctggcacgc cgcgccgcag gcaaggcaga 3480
agccagatgg ttgttcaaga cgatctacga acgcagtggc agcgccggag agttcaagaa 3540
gttctgtttc accgtgcgca agctgatcgg gtcaaatgac ctgccggagt acgatttgaa 3600
ggaggaggcg gggcaggctg gcccgatcct agtcatgcgc taccgcaacc tgatcgaggg 3660
cgaagcatcc gccggttcct aatgtacgga gcagatgcta gggcaaattg ccctagcagg 3720
ggaaaaaggt cgaaaaggtc tctttcctgt ggatagcacg tacattggga acccaaagcc 3780
gtacattggg aaccggaacc cgtacattgg gaacccaaag ccgtacattg ggaaccggtc 3840
acacatgtaa gtgactgata taaaagagaa aaaaggcgat ttttccgcct aaaactcttt 3900
aaaacttatt aaaactctta aaacccgcct ggcctgtgca taactgtctg gccagcgcac 3960
agccgaagag ctgcaaaaag cgcctaccct tcggtcgctg cgctccctac gccccgccgc 4020
ttcgcgtcgg cctatcgcgg ccgctggccg ctcaaaaatg gctggcctac ggccaggcaa 4080
tctaccaggg cgcggacaag ccgcgccgtc gccactcgac cgccggcgcc cacatcaagg 4140
caccctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg 4200
agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt 4260
cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag 4320
tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg 4380
gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct cttccgcttc 4440
ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc 4500
aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc 4560
aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag 4620
gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc 4680
gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt 4740
tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct 4800
ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg 4860
ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct 4920
tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat 4980
tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg 5040
ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa 5100
aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt 5160
ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc 5220
tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgcattc 5280
taggtactaa aacaattcat ccagtaaaat ataatatttt attttctccc aatcaggctt 5340
gatccccagt aagtcaaaaa atagctcgac atactgttct tccccgatat cctccctgat 5400
cgaccggacg cagaaggcaa tgtcatacca cttgtccgcc ctgccgcttc tcccaagatc 5460
aataaagcca cttactttgc catctttcac aaagatgttg ctgtctccca ggtcgccgtg 5520
ggaaaagaca agttcctctt cgggcttttc cgtctttaaa aaatcataca gctcgcgcgg 5580
atctttaaat ggagtgtctt cttcccagtt ttcgcaatcc acatcggcca gatcgttatt 5640
cagtaagtaa tccaattcgg ctaagcggct gtctaagcta ttcgtatagg gacaatccga 5700
tatgtcgatg gagtgaaaga gcctgatgca ctccgcatac agctcgataa tcttttcagg 5760
gctttgttca tcttcatact cttccgagca aaggacgcca tcggcctcac tcatgagcag 5820
attgctccag ccatcatgcc gttcaaagtg caggaccttt ggaacaggca gctttccttc 5880
cagccatagc atcatgtcct tttcccgttc cacatcatag gtggtccctt tataccggct 5940
gtccgtcatt tttaaatata ggttttcatt ttctcccacc agcttatata ccttagcagg 6000
agacattcct tccgtatctt ttacgcagcg gtatttttcg atcagttttt tcaattccgg 6060
tgatattctc attttagcca tttattattt ccttcctctt ttctacagta tttaaagata 6120
ccccaagaag ctaattataa caagacgaac tccaattcac tgttccttgc attctaaaac 6180
cttaaatacc agaaaacagc tttttcaaag ttgttttcaa agttggcgta taacatagta 6240
tcgacggagc cgattttgaa accgcggtga tcacaggcag caacgctctg tcatcgttac 6300
aatcaacatg ctaccctccg cgagatcatc cgtgtttcaa acccggcagc ttagttgccg 6360
ttcttccgaa tagcatcggt aacatgagca aagtctgccg ccttacaacg gctctcccgc 6420
tgacgccgtc ccggactgat gggctgcctg tatcgagtgg tgattttgtg ccgagctgcc 6480
ggtcggggag ctgttggctg gctggtggca ggatatattg tggtgtaaac aaattgacgc 6540
ttagacaact taataacaca ttgcggacgt ttttaatgta ctgaattaac gccgaattaa 6600
ttcgggggat ctggatttta gtactggatt ttggttttag gaattagaaa ttttattgat 6660
agaagtattt tacaaataca aatacatact aagggtttct tatatgctca acacatgagc 6720
gaaaccctat aggaacccta attcccttat ctgggaacta ctcacacatt attatggaga 6780
aactcgagct tgtcgatcga cagatccggt cggcatctac tctatttctt tgccctcgga 6840
cgagtgctgg ggcgtcggtt tccactatcg gcgagtactt ctacacagcc atcggtccag 6900
acggccgcgc ttctgcgggc gatttgtgta cgcccgacag tcccggctcc ggatcggacg 6960
attgcgtcgc atcgaccctg cgcccaagct gcatcatcga aattgccgtc aaccaagctc 7020
tgatagagtt ggtcaagacc aatgcggagc atatacgccc ggagtcgtgg cgatcctgca 7080
agctccggat gcctccgctc gaagtagcgc gtctgctgct ccatacaagc caaccacggc 7140
ctccagaaga agatgttggc gacctcgtat tgggaatccc cgaacatcgc ctcgctccag 7200
tcaatgaccg ctgttatgcg gccattgtcc gtcaggacat tgttggagcc gaaatccgcg 7260
tgcacgaggt gccggacttc ggggcagtcc tcggcccaaa gcatcagctc atcgagagcc 7320
tgcgcgacgg acgcactgac ggtgtcgtcc atcacagttt gccagtgata cacatgggga 7380
tcagcaatcg cgcatatgaa atcacgccat gtagtgtatt gaccgattcc ttgcggtccg 7440
aatgggccga acccgctcgt ctggctaaga tcggccgcag cgatcgcatc catagcctcc 7500
gcgaccggtt gtagaacagc gggcagttcg gtttcaggca ggtcttgcaa cgtgacaccc 7560
tgtgcacggc gggagatgca ataggtcagg ctctcgctaa actccccaat gtcaagcact 7620
tccggaatcg ggagcgcggc cgatgcaaag tgccgataaa cataacgatc tttgtagaaa 7680
ccatcggcgc agctatttac ccgcaggaca tatccacgcc ctcctacatc gaagctgaaa 7740
gcacgagatt cttcgccctc cgagagctgc atcaggtcgg agacgctgtc gaacttttcg 7800
atcagaaact tctcgacaga cgtcgcggtg agttcaggct ttttcatatc tcattgcccc 7860
ccgggatctg cgaaagctcg agagagatag atttgtagag agagactggt gatttcagcg 7920
tgtcctctcc aaatgaaatg aacttcctta tatagaggaa ggtcttgcga aggatagtgg 7980
gattgtgcgt catcccttac gtcagtggag atatcacatc aatccacttg ctttgaagac 8040
gtggttggaa cgtcttcttt ttccacgatg ctcctcgtgg gtgggggtcc atctttggga 8100
ccactgtcgg cagaggcatc ttgaacgata gcctttcctt tatcgcaatg atggcatttg 8160
taggtgccac cttccttttc tactgtcctt ttgatgaagt gacagatagc tgggcaatgg 8220
aatccgagga ggtttcccga tattaccctt tgttgaaaag tctcaatagc cctttggtct 8280
tctgagactg tatctttgat attcttggag tagacgagag tgtcgtgctc caccatgtta 8340
tcacatcaat ccacttgctt tgaagacgtg gttggaacgt cttctttttc cacgatgctc 8400
ctcgtgggtg ggggtccatc tttgggacca ctgtcggcag aggcatcttg aacgatagcc 8460
tttcctttat cgcaatgatg gcatttgtag gtgccacctt ccttttctac tgtccttttg 8520
atgaagtgac agatagctgg gcaatggaat ccgaggaggt ttcccgatat taccctttgt 8580
tgaaaagtct caatagccct ttggtcttct gagactgtat ctttgatatt cttggagtag 8640
acgagagtgt cgtgctccac catgttggca agctgctcta gccaatacgc aaaccgcctc 8700
tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag 8760
cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt 8820
tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca 8880
caggaaacag ctatgaccat gattacgaat tcccgatcta gtaacataga tgacaccgcg 8940
cgcgataatt tatcctagtt tgcgcgctat attttgtttt ctatcgcgta ttaaatgtat 9000
aattgcggga ctctaatcat aaaaacccat ctcataaata acgtcatgca ttacatgtta 9060
attattacat gcttaacgta attcaacaga aattatatga taatcatcgc aagaccggca 9120
acaggattca atcttaagaa actttattgc caaatgtttg aacgatcggg gaaattcgag 9180
ctctaagctt aggcgccggt ggagtggcgg ccctcggcgc gctcgtactg ttccacgatg 9240
gtgtagtcct cgttgtggga ggtgatgtcc agcttgatgt cggtcttgta ggcgccgggc 9300
agctgcacgg gcttcttggc catgtaggtg gtcttgacct cggcgtcgta gtggccgccg 9360
tccttcagct tcagcctcat cttgatctcg cccttcaggg cgccgtcctc ggggtacatc 9420
cgctcggtgg aggcctccca gcccatggtc ttcttctgca ttacggggcc gtcggagggg 9480
aagttggtgc cgcgcagctt caccttgtag atgaactcgc cgtcctgcag ggaggagtcc 9540
tgggtcacgg tcaccacgcc gccgtcctcg aagttcatca cgcgctccca cttgaagccc 9600
tcggggaagg acagcttcaa gtagtcgggg atgtcggcgg ggtgcttcac gtaggccttg 9660
gagccgtact ggaactgagg ggacaggatg tcccaggcga agggcagggg gccgcccttg 9720
gtcaccttca gcttggcggt ctgggtgccc tcgtaggggc ggccctcgcc ctcgccctcg 9780
atctcgaact cgtggccgtt cacggagccc tccatgcgca ccttgaagcg catgaactcc 9840
ttgatgacgt cctcggagga ggccatagct gcaaacataa caaatatacc tcagaatcaa 9900
taaactaggc taaaagggaa ctgaagagaa tcaacagact gtagtctctg agcagtaact 9960
acacaaaaac atatgataga acaatctggt aaaaacatat gcagtatgtt ctacaccaca 10020
aatttattac atacaacaat tacatctatc agcgtgaaga acgcatgtca aatagtttct 10080
ttcaattttt catacaccaa aacaaaattc atgggctaaa accccaagac tttaaattgc 10140
ataaatactg acaagtggca acaaaagctg attttaaggg tatgtaaacc taccagtaag 10200
gtggacgttg acagggcaaa caaattagaa ttttgcactg atactaaaat gggcttccaa 10260
accctaatag gcactagatc agactgcaca cacatacaac tactaatggg aaaactaggt 10320
cagcatatac attcaacgcc gtaaattggc atagaaaact tcatcatcac agatgaaacc 10380
aagcagcttg tatttcttat gtggttataa aggggagcac ttgccattgc tgttacaaga 10440
catacattta tgtagattaa taatgatttt ccatggaatc cacaagcatt tgtaagcgaa 10500
aacgaagcaa acattacaaa attcccaagt atcactaaag cagaatagca gatgcaagta 10560
acatggcacc tattagtaac ctaagccgta gcaaacttca cactccacgt ctccagaaca 10620
aataaaagct ctgcaacgac ctaaagttat gcgaaaagtg caaaatcccc aacaagcgcc 10680
acttcgtcat gtgaaccgaa ccagccaaaa tctcaaacct cccctcccct tctcgattct 10740
cataacgcaa tagaagtata gagaagggat tgcttcacgg gacgcacctt tgatcccgag 10800
ggacgtctgg ccggatccga ccgcggtgag cgcgtgctcg atctgcacca gcttccccga 10860
cgggctgcaa acgaaatcgc cacaaaaacc acagatgacg aaaccgaaac cgcaccagat 10920
cacgcccgat ctccgggcga gggaggcgca aggcagcaca tacctgaagg tggtgaggga 10980
gaaggagtac tggctgtctc ccatggccgc cgacgcgcag gcctacctgt gagctgctgc 11040
tcttcgtgtc tcgcggcgga agaggaggcg accgagcgag cgattgcggc gagacggcgt 11100
gggaagggag agggaggcag agcacaactc gccaaggaag aagaggagag agagaaagag 11160
actggatgga tggattgctc cgcaagaagg tggtgggtct tgggccggaa atctggccca 11220
tctgttattg ggtcagccca tgactcaccc acctccccgt cacgtcgtct tcctcctcat 11280
aacttcttcc ccacgggcca acggctgcgc gtcttcccgt tcgtcctcct gtccggtccg 11340
agtccctctg acgacgcgac tttccccacg ccgggatcga gctccgggga ggggagctgc 11400
gagctggtac gtacgcttga tttcttccct tgccaccgcg atattccgat gtatccgtga 11460
tgtgtcgttc atgggtagat cggctaggga tttggcggcg ttgaggaaag cgatggctgc 11520
cgggtggggg gaattgttgt attagactct cgaagctgat atctggtgtg gatttgcggt 11580
ggaagccatg gtcggaggcc cagtgccttc cgggagttgt tgggttatcg ggagcccgga 11640
gggcagtgcg gctcgccgac tgctgccatg atccccgtcg gcaaaatctc cagattgggg 11700
gagatggggg gtaggaacca tttggattag tagtgtatac acggggtttc gtttattata 11760
tttagagcgc tctacatgta gacagcacac agcagcttta acgtatggat ggtgtatcgc 11820
agggaatgta tctcgaagac attggattag gctattatta catttagaga accagattgg 11880
gtttgcgata actagaaatt gagtctgtta tgaatttatt gatggagcgg acagggaatt 11940
acgattacta tctgtttagc ttttcagttg tgaatgatga ttatttcaat ttagctaatc 12000
aaggttcctc aatcaaaccg ttctaacttt ttctatttat gctcttggtt acattttgac 12060
agaaacactt tgtcatatca gtcaacagta gagaggagac gagatggctc ctaagaagaa 12120
gagaaaggtt ataacaatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat 12180
cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga 12240
gggcgatgcc acctacggca agctgaccct gaagctgatc tgcaccaccg gcaagctgcc 12300
cgtgccctgg cccaccctcg tgaccaccct gggctacggc ctgcagtgct tcgcccgcta 12360
ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca 12420
ggagcgcacc atcttcttca aggacgacgg caactacaag acacgcgccg aggtgaagtt 12480
cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg 12540
caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcaccgc 12600
cgacaagcag aagaacggca tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg 12660
cggcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct 12720
gctgcccgac aaccactacc tgagctacca gtccgccctg agcaaagacc ccaacgagaa 12780
gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga 12840
cgaggcagct agatccacct gagtcgagtc gacggatata tgaagatgaa gatgaaatat 12900
ttggtgtgtc aaataaaaag cttgtgtgct taagtttgtg tttttttctt ggcttgttgt 12960
gttatgaatt tgtggctttt tctaatatta aatgaatgta agatcacatt ataatgaata 13020
aacaaatgtt tctataatcc attgtgaatg ttttgttgga tctcttctgc agcatataac 13080
tactgtatgt gctatggtat ggactatgga atatgattaa agataagg 13128
<210> 4
<211> 11218
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 4
agcttggcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa 60
cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc 120
accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgctagag cagcttgagc 180
ttggatcaga ttgtcgtttc ccgccttcag tttaaactat cagtgtttga caggatatat 240
tggcgggtaa acctaagaga aaagagcgtt tattagaata acggatattt aaaagggcgt 300
gaaaaggttt atccgttcgt ccatttgtat gtgcatgcca accacagggt tcccctcggg 360
atcaaagtac tttgatccaa cccctccgct gctatagtgc agtcggcttc tgacgttcag 420
tgcagccgtc ttctgaaaac gacatgtcgc acaagtccta agttacgcga caggctgccg 480
ccctgccctt ttcctggcgt tttcttgtcg cgtgttttag tcgcataaag tagaatactt 540
gcgactagaa ccggagacat tacgccatga acaagagcgc cgccgctggc ctgctgggct 600
atgcccgcgt cagcaccgac gaccaggact tgaccaacca acgggccgaa ctgcacgcgg 660
ccggctgcac caagctgttt tccgagaaga tcaccggcac caggcgcgac cgcccggagc 720
tggccaggat gcttgaccac ctacgccctg gcgacgttgt gacagtgacc aggctagacc 780
gcctggcccg cagcacccgc gacctactgg acattgccga gcgcatccag gaggccggcg 840
cgggcctgcg tagcctggca gagccgtggg ccgacaccac cacgccggcc ggccgcatgg 900
tgttgaccgt gttcgccggc attgccgagt tcgagcgttc cctaatcatc gaccgcaccc 960
ggagcgggcg cgaggccgcc aaggcccgag gcgtgaagtt tggcccccgc cctaccctca 1020
ccccggcaca gatcgcgcac gcccgcgagc tgatcgacca ggaaggccgc accgtgaaag 1080
aggcggctgc actgcttggc gtgcatcgct cgaccctgta ccgcgcactt gagcgcagcg 1140
aggaagtgac gcccaccgag gccaggcggc gcggtgcctt ccgtgaggac gcattgaccg 1200
aggccgacgc cctggcggcc gccgagaatg aacgccaaga ggaacaagca tgaaaccgca 1260
ccaggacggc caggacgaac cgtttttcat taccgaagag atcgaggcgg agatgatcgc 1320
ggccgggtac gtgttcgagc cgcccgcgca cgtctcaacc gtgcggctgc atgaaatcct 1380
ggccggtttg tctgatgcca agctggcggc ctggccggcc agcttggccg ctgaagaaac 1440
cgagcgccgc cgtctaaaaa ggtgatgtgt atttgagtaa aacagcttgc gtcatgcggt 1500
cgctgcgtat atgatgcgat gagtaaataa acaaatacgc aaggggaacg catgaaggtt 1560
atcgctgtac ttaaccagaa aggcgggtca ggcaagacga ccatcgcaac ccatctagcc 1620
cgcgccctgc aactcgccgg ggccgatgtt ctgttagtcg attccgatcc ccagggcagt 1680
gcccgcgatt gggcggccgt gcgggaagat caaccgctaa ccgttgtcgg catcgaccgc 1740
ccgacgattg accgcgacgt gaaggccatc ggccggcgcg acttcgtagt gatcgacgga 1800
gcgccccagg cggcggactt ggctgtgtcc gcgatcaagg cagccgactt cgtgctgatt 1860
ccggtgcagc caagccctta cgacatatgg gccaccgccg acctggtgga gctggttaag 1920
cagcgcattg aggtcacgga tggaaggcta caagcggcct ttgtcgtgtc gcgggcgatc 1980
aaaggcacgc gcatcggcgg tgaggttgcc gaggcgctgg ccgggtacga gctgcccatt 2040
cttgagtccc gtatcacgca gcgcgtgagc tacccaggca ctgccgccgc cggcacaacc 2100
gttcttgaat cagaacccga gggcgacgct gcccgcgagg tccaggcgct ggccgctgaa 2160
attaaatcaa aactcatttg agttaatgag gtaaagagaa aatgagcaaa agcacaaaca 2220
cgctaagtgc cggccgtccg agcgcacgca gcagcaaggc tgcaacgttg gccagcctgg 2280
cagacacgcc agccatgaag cgggtcaact ttcagttgcc ggcggaggat cacaccaagc 2340
tgaagatgta cgcggtacgc caaggcaaga ccattaccga gctgctatct gaatacatcg 2400
cgcagctacc agagtaaatg agcaaatgaa taaatgagta gatgaatttt agcggctaaa 2460
ggaggcggca tggaaaatca agaacaacca ggcaccgacg ccgtggaatg ccccatgtgt 2520
ggaggaacgg gcggttggcc aggcgtaagc ggctgggttg tctgccggcc ctgcaatggc 2580
actggaaccc ccaagcccga ggaatcggcg tgacggtcgc aaaccatccg gcccggtaca 2640
aatcggcgcg gcgctgggtg atgacctggt ggagaagttg aaggccgcgc aggccgccca 2700
gcggcaacgc atcgaggcag aagcacgccc cggtgaatcg tggcaagcgg ccgctgatcg 2760
aatccgcaaa gaatcccggc aaccgccggc agccggtgcg ccgtcgatta ggaagccgcc 2820
caagggcgac gagcaaccag attttttcgt tccgatgctc tatgacgtgg gcacccgcga 2880
tagtcgcagc atcatggacg tggccgtttt ccgtctgtcg aagcgtgacc gacgagctgg 2940
cgaggtgatc cgctacgagc ttccagacgg gcacgtagag gtttccgcag ggccggccgg 3000
catggccagt gtgtgggatt acgacctggt actgatggcg gtttcccatc taaccgaatc 3060
catgaaccga taccgggaag ggaagggaga caagcccggc cgcgtgttcc gtccacacgt 3120
tgcggacgta ctcaagttct gccggcgagc cgatggcgga aagcagaaag acgacctggt 3180
agaaacctgc attcggttaa acaccacgca cgttgccatg cagcgtacga agaaggccaa 3240
gaacggccgc ctggtgacgg tatccgaggg tgaagccttg attagccgct acaagatcgt 3300
aaagagcgaa accgggcggc cggagtacat cgagatcgag ctagctgatt ggatgtaccg 3360
cgagatcaca gaaggcaaga acccggacgt gctgacggtt caccccgatt actttttgat 3420
cgatcccggc atcggccgtt ttctctaccg cctggcacgc cgcgccgcag gcaaggcaga 3480
agccagatgg ttgttcaaga cgatctacga acgcagtggc agcgccggag agttcaagaa 3540
gttctgtttc accgtgcgca agctgatcgg gtcaaatgac ctgccggagt acgatttgaa 3600
ggaggaggcg gggcaggctg gcccgatcct agtcatgcgc taccgcaacc tgatcgaggg 3660
cgaagcatcc gccggttcct aatgtacgga gcagatgcta gggcaaattg ccctagcagg 3720
ggaaaaaggt cgaaaaggtc tctttcctgt ggatagcacg tacattggga acccaaagcc 3780
gtacattggg aaccggaacc cgtacattgg gaacccaaag ccgtacattg ggaaccggtc 3840
acacatgtaa gtgactgata taaaagagaa aaaaggcgat ttttccgcct aaaactcttt 3900
aaaacttatt aaaactctta aaacccgcct ggcctgtgca taactgtctg gccagcgcac 3960
agccgaagag ctgcaaaaag cgcctaccct tcggtcgctg cgctccctac gccccgccgc 4020
ttcgcgtcgg cctatcgcgg ccgctggccg ctcaaaaatg gctggcctac ggccaggcaa 4080
tctaccaggg cgcggacaag ccgcgccgtc gccactcgac cgccggcgcc cacatcaagg 4140
caccctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg 4200
agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt 4260
cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag 4320
tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg 4380
gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct cttccgcttc 4440
ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc 4500
aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc 4560
aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag 4620
gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc 4680
gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt 4740
tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct 4800
ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg 4860
ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct 4920
tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat 4980
tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg 5040
ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa 5100
aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt 5160
ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc 5220
tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgcattc 5280
taggtactaa aacaattcat ccagtaaaat ataatatttt attttctccc aatcaggctt 5340
gatccccagt aagtcaaaaa atagctcgac atactgttct tccccgatat cctccctgat 5400
cgaccggacg cagaaggcaa tgtcatacca cttgtccgcc ctgccgcttc tcccaagatc 5460
aataaagcca cttactttgc catctttcac aaagatgttg ctgtctccca ggtcgccgtg 5520
ggaaaagaca agttcctctt cgggcttttc cgtctttaaa aaatcataca gctcgcgcgg 5580
atctttaaat ggagtgtctt cttcccagtt ttcgcaatcc acatcggcca gatcgttatt 5640
cagtaagtaa tccaattcgg ctaagcggct gtctaagcta ttcgtatagg gacaatccga 5700
tatgtcgatg gagtgaaaga gcctgatgca ctccgcatac agctcgataa tcttttcagg 5760
gctttgttca tcttcatact cttccgagca aaggacgcca tcggcctcac tcatgagcag 5820
attgctccag ccatcatgcc gttcaaagtg caggaccttt ggaacaggca gctttccttc 5880
cagccatagc atcatgtcct tttcccgttc cacatcatag gtggtccctt tataccggct 5940
gtccgtcatt tttaaatata ggttttcatt ttctcccacc agcttatata ccttagcagg 6000
agacattcct tccgtatctt ttacgcagcg gtatttttcg atcagttttt tcaattccgg 6060
tgatattctc attttagcca tttattattt ccttcctctt ttctacagta tttaaagata 6120
ccccaagaag ctaattataa caagacgaac tccaattcac tgttccttgc attctaaaac 6180
cttaaatacc agaaaacagc tttttcaaag ttgttttcaa agttggcgta taacatagta 6240
tcgacggagc cgattttgaa accgcggtga tcacaggcag caacgctctg tcatcgttac 6300
aatcaacatg ctaccctccg cgagatcatc cgtgtttcaa acccggcagc ttagttgccg 6360
ttcttccgaa tagcatcggt aacatgagca aagtctgccg ccttacaacg gctctcccgc 6420
tgacgccgtc ccggactgat gggctgcctg tatcgagtgg tgattttgtg ccgagctgcc 6480
ggtcggggag ctgttggctg gctggtggca ggatatattg tggtgtaaac aaattgacgc 6540
ttagacaact taataacaca ttgcggacgt ttttaatgta ctgaattaac gccgaattaa 6600
ttcgggggat ctggatttta gtactggatt ttggttttag gaattagaaa ttttattgat 6660
agaagtattt tacaaataca aatacatact aagggtttct tatatgctca acacatgagc 6720
gaaaccctat aggaacccta attcccttat ctgggaacta ctcacacatt attatggaga 6780
aactcgagct tgtcgatcga cagatccggt cggcatctac tctatttctt tgccctcgga 6840
cgagtgctgg ggcgtcggtt tccactatcg gcgagtactt ctacacagcc atcggtccag 6900
acggccgcgc ttctgcgggc gatttgtgta cgcccgacag tcccggctcc ggatcggacg 6960
attgcgtcgc atcgaccctg cgcccaagct gcatcatcga aattgccgtc aaccaagctc 7020
tgatagagtt ggtcaagacc aatgcggagc atatacgccc ggagtcgtgg cgatcctgca 7080
agctccggat gcctccgctc gaagtagcgc gtctgctgct ccatacaagc caaccacggc 7140
ctccagaaga agatgttggc gacctcgtat tgggaatccc cgaacatcgc ctcgctccag 7200
tcaatgaccg ctgttatgcg gccattgtcc gtcaggacat tgttggagcc gaaatccgcg 7260
tgcacgaggt gccggacttc ggggcagtcc tcggcccaaa gcatcagctc atcgagagcc 7320
tgcgcgacgg acgcactgac ggtgtcgtcc atcacagttt gccagtgata cacatgggga 7380
tcagcaatcg cgcatatgaa atcacgccat gtagtgtatt gaccgattcc ttgcggtccg 7440
aatgggccga acccgctcgt ctggctaaga tcggccgcag cgatcgcatc catagcctcc 7500
gcgaccggtt gtagaacagc gggcagttcg gtttcaggca ggtcttgcaa cgtgacaccc 7560
tgtgcacggc gggagatgca ataggtcagg ctctcgctaa actccccaat gtcaagcact 7620
tccggaatcg ggagcgcggc cgatgcaaag tgccgataaa cataacgatc tttgtagaaa 7680
ccatcggcgc agctatttac ccgcaggaca tatccacgcc ctcctacatc gaagctgaaa 7740
gcacgagatt cttcgccctc cgagagctgc atcaggtcgg agacgctgtc gaacttttcg 7800
atcagaaact tctcgacaga cgtcgcggtg agttcaggct ttttcatatc tcattgcccc 7860
ccgggatctg cgaaagctcg agagagatag atttgtagag agagactggt gatttcagcg 7920
tgtcctctcc aaatgaaatg aacttcctta tatagaggaa ggtcttgcga aggatagtgg 7980
gattgtgcgt catcccttac gtcagtggag atatcacatc aatccacttg ctttgaagac 8040
gtggttggaa cgtcttcttt ttccacgatg ctcctcgtgg gtgggggtcc atctttggga 8100
ccactgtcgg cagaggcatc ttgaacgata gcctttcctt tatcgcaatg atggcatttg 8160
taggtgccac cttccttttc tactgtcctt ttgatgaagt gacagatagc tgggcaatgg 8220
aatccgagga ggtttcccga tattaccctt tgttgaaaag tctcaatagc cctttggtct 8280
tctgagactg tatctttgat attcttggag tagacgagag tgtcgtgctc caccatgtta 8340
tcacatcaat ccacttgctt tgaagacgtg gttggaacgt cttctttttc cacgatgctc 8400
ctcgtgggtg ggggtccatc tttgggacca ctgtcggcag aggcatcttg aacgatagcc 8460
tttcctttat cgcaatgatg gcatttgtag gtgccacctt ccttttctac tgtccttttg 8520
atgaagtgac agatagctgg gcaatggaat ccgaggaggt ttcccgatat taccctttgt 8580
tgaaaagtct caatagccct ttggtcttct gagactgtat ctttgatatt cttggagtag 8640
acgagagtgt cgtgctccac catgttggca agctgctcta gccaatacgc aaaccgcctc 8700
tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag 8760
cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt 8820
tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca 8880
caggaaacag ctatgaccat gattacgaat tcccgatcta gtaacataga tgacaccgcg 8940
cgcgataatt tatcctagtt tgcgcgctat attttgtttt ctatcgcgta ttaaatgtat 9000
aattgcggga ctctaatcat aaaaacccat ctcataaata acgtcatgca ttacatgtta 9060
attattacat gcttaacgta attcaacaga aattatatga taatcatcgc aagaccggca 9120
acaggattca atcttaagaa actttattgc caaatgtttg aacgatcggg gaaattcgag 9180
ctctaagctt aggcgccggt ggagtggcgg ccctcggcgc gctcgtactg ttccacgatg 9240
gtgtagtcct cgttgtggga ggtgatgtcc agcttgatgt cggtcttgta ggcgccgggc 9300
agctgcacgg gcttcttggc catgtaggtg gtcttgacct cggcgtcgta gtggccgccg 9360
tccttcagct tcagcctcat cttgatctcg cccttcaggg cgccgtcctc ggggtacatc 9420
cgctcggtgg aggcctccca gcccatggtc ttcttctgca ttacggggcc gtcggagggg 9480
aagttggtgc cgcgcagctt caccttgtag atgaactcgc cgtcctgcag ggaggagtcc 9540
tgggtcacgg tcaccacgcc gccgtcctcg aagttcatca cgcgctccca cttgaagccc 9600
tcggggaagg acagcttcaa gtagtcgggg atgtcggcgg ggtgcttcac gtaggccttg 9660
gagccgtact ggaactgagg ggacaggatg tcccaggcga agggcagggg gccgcccttg 9720
gtcaccttca gcttggcggt ctgggtgccc tcgtaggggc ggccctcgcc ctcgccctcg 9780
atctcgaact cgtggccgtt cacggagccc tccatgcgca ccttgaagcg catgaactcc 9840
ttgatgacgt cctcggagga ggccattgtt ataacctttc tcttcttctt aggagccatg 9900
gatccttctc tccaaatgaa atgaacttcc ttatatagag gaagggtctt gcgaaatgga 9960
gtcaaagatt caaatagagg acctaacaga actcgccgta aagactggcg aacagttcat 10020
acagagtctc ttacgactca atgacaagaa gaaaatcttc gtcaacatgg tggagcacga 10080
cacacttgtc tactccaaaa atatcaaaga tacagtctca gaagaccaaa gggcaatttc 10140
gcaagaccct tcctctatat aaggaagttc atttcatttg gagagaagga tccatggctc 10200
ctaagaagaa gagaaaggtt ataacaatgg tgagcaaggg cgaggagctg ttcaccgggg 10260
tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg 10320
gcgagggcga gggcgatgcc acctacggca agctgaccct gaagctgatc tgcaccaccg 10380
gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gggctacggc ctgcagtgct 10440
tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag 10500
gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acacgcgccg 10560
aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca 10620
aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct 10680
atatcaccgc cgacaagcag aagaacggca tcaaggccaa cttcaagatc cgccacaaca 10740
tcgaggacgg cggcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg 10800
gccccgtgct gctgcccgac aaccactacc tgagctacca gtccgccctg agcaaagacc 10860
ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc 10920
tcggcatgga cgaggcagct agatccacct gagtcgagtc gacggatata tgaagatgaa 10980
gatgaaatat ttggtgtgtc aaataaaaag cttgtgtgct taagtttgtg tttttttctt 11040
ggcttgttgt gttatgaatt tgtggctttt tctaatatta aatgaatgta agatcacatt 11100
ataatgaata aacaaatgtt tctataatcc attgtgaatg ttttgttgga tctcttctgc 11160
agcatataac tactgtatgt gctatggtat ggactatgga atatgattaa agataagg 11218
<210> 5
<211> 11118
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 5
agcttggcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa 60
cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc 120
accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgctagag cagcttgagc 180
ttggatcaga ttgtcgtttc ccgccttcag tttaaactat cagtgtttga caggatatat 240
tggcgggtaa acctaagaga aaagagcgtt tattagaata acggatattt aaaagggcgt 300
gaaaaggttt atccgttcgt ccatttgtat gtgcatgcca accacagggt tcccctcggg 360
atcaaagtac tttgatccaa cccctccgct gctatagtgc agtcggcttc tgacgttcag 420
tgcagccgtc ttctgaaaac gacatgtcgc acaagtccta agttacgcga caggctgccg 480
ccctgccctt ttcctggcgt tttcttgtcg cgtgttttag tcgcataaag tagaatactt 540
gcgactagaa ccggagacat tacgccatga acaagagcgc cgccgctggc ctgctgggct 600
atgcccgcgt cagcaccgac gaccaggact tgaccaacca acgggccgaa ctgcacgcgg 660
ccggctgcac caagctgttt tccgagaaga tcaccggcac caggcgcgac cgcccggagc 720
tggccaggat gcttgaccac ctacgccctg gcgacgttgt gacagtgacc aggctagacc 780
gcctggcccg cagcacccgc gacctactgg acattgccga gcgcatccag gaggccggcg 840
cgggcctgcg tagcctggca gagccgtggg ccgacaccac cacgccggcc ggccgcatgg 900
tgttgaccgt gttcgccggc attgccgagt tcgagcgttc cctaatcatc gaccgcaccc 960
ggagcgggcg cgaggccgcc aaggcccgag gcgtgaagtt tggcccccgc cctaccctca 1020
ccccggcaca gatcgcgcac gcccgcgagc tgatcgacca ggaaggccgc accgtgaaag 1080
aggcggctgc actgcttggc gtgcatcgct cgaccctgta ccgcgcactt gagcgcagcg 1140
aggaagtgac gcccaccgag gccaggcggc gcggtgcctt ccgtgaggac gcattgaccg 1200
aggccgacgc cctggcggcc gccgagaatg aacgccaaga ggaacaagca tgaaaccgca 1260
ccaggacggc caggacgaac cgtttttcat taccgaagag atcgaggcgg agatgatcgc 1320
ggccgggtac gtgttcgagc cgcccgcgca cgtctcaacc gtgcggctgc atgaaatcct 1380
ggccggtttg tctgatgcca agctggcggc ctggccggcc agcttggccg ctgaagaaac 1440
cgagcgccgc cgtctaaaaa ggtgatgtgt atttgagtaa aacagcttgc gtcatgcggt 1500
cgctgcgtat atgatgcgat gagtaaataa acaaatacgc aaggggaacg catgaaggtt 1560
atcgctgtac ttaaccagaa aggcgggtca ggcaagacga ccatcgcaac ccatctagcc 1620
cgcgccctgc aactcgccgg ggccgatgtt ctgttagtcg attccgatcc ccagggcagt 1680
gcccgcgatt gggcggccgt gcgggaagat caaccgctaa ccgttgtcgg catcgaccgc 1740
ccgacgattg accgcgacgt gaaggccatc ggccggcgcg acttcgtagt gatcgacgga 1800
gcgccccagg cggcggactt ggctgtgtcc gcgatcaagg cagccgactt cgtgctgatt 1860
ccggtgcagc caagccctta cgacatatgg gccaccgccg acctggtgga gctggttaag 1920
cagcgcattg aggtcacgga tggaaggcta caagcggcct ttgtcgtgtc gcgggcgatc 1980
aaaggcacgc gcatcggcgg tgaggttgcc gaggcgctgg ccgggtacga gctgcccatt 2040
cttgagtccc gtatcacgca gcgcgtgagc tacccaggca ctgccgccgc cggcacaacc 2100
gttcttgaat cagaacccga gggcgacgct gcccgcgagg tccaggcgct ggccgctgaa 2160
attaaatcaa aactcatttg agttaatgag gtaaagagaa aatgagcaaa agcacaaaca 2220
cgctaagtgc cggccgtccg agcgcacgca gcagcaaggc tgcaacgttg gccagcctgg 2280
cagacacgcc agccatgaag cgggtcaact ttcagttgcc ggcggaggat cacaccaagc 2340
tgaagatgta cgcggtacgc caaggcaaga ccattaccga gctgctatct gaatacatcg 2400
cgcagctacc agagtaaatg agcaaatgaa taaatgagta gatgaatttt agcggctaaa 2460
ggaggcggca tggaaaatca agaacaacca ggcaccgacg ccgtggaatg ccccatgtgt 2520
ggaggaacgg gcggttggcc aggcgtaagc ggctgggttg tctgccggcc ctgcaatggc 2580
actggaaccc ccaagcccga ggaatcggcg tgacggtcgc aaaccatccg gcccggtaca 2640
aatcggcgcg gcgctgggtg atgacctggt ggagaagttg aaggccgcgc aggccgccca 2700
gcggcaacgc atcgaggcag aagcacgccc cggtgaatcg tggcaagcgg ccgctgatcg 2760
aatccgcaaa gaatcccggc aaccgccggc agccggtgcg ccgtcgatta ggaagccgcc 2820
caagggcgac gagcaaccag attttttcgt tccgatgctc tatgacgtgg gcacccgcga 2880
tagtcgcagc atcatggacg tggccgtttt ccgtctgtcg aagcgtgacc gacgagctgg 2940
cgaggtgatc cgctacgagc ttccagacgg gcacgtagag gtttccgcag ggccggccgg 3000
catggccagt gtgtgggatt acgacctggt actgatggcg gtttcccatc taaccgaatc 3060
catgaaccga taccgggaag ggaagggaga caagcccggc cgcgtgttcc gtccacacgt 3120
tgcggacgta ctcaagttct gccggcgagc cgatggcgga aagcagaaag acgacctggt 3180
agaaacctgc attcggttaa acaccacgca cgttgccatg cagcgtacga agaaggccaa 3240
gaacggccgc ctggtgacgg tatccgaggg tgaagccttg attagccgct acaagatcgt 3300
aaagagcgaa accgggcggc cggagtacat cgagatcgag ctagctgatt ggatgtaccg 3360
cgagatcaca gaaggcaaga acccggacgt gctgacggtt caccccgatt actttttgat 3420
cgatcccggc atcggccgtt ttctctaccg cctggcacgc cgcgccgcag gcaaggcaga 3480
agccagatgg ttgttcaaga cgatctacga acgcagtggc agcgccggag agttcaagaa 3540
gttctgtttc accgtgcgca agctgatcgg gtcaaatgac ctgccggagt acgatttgaa 3600
ggaggaggcg gggcaggctg gcccgatcct agtcatgcgc taccgcaacc tgatcgaggg 3660
cgaagcatcc gccggttcct aatgtacgga gcagatgcta gggcaaattg ccctagcagg 3720
ggaaaaaggt cgaaaaggtc tctttcctgt ggatagcacg tacattggga acccaaagcc 3780
gtacattggg aaccggaacc cgtacattgg gaacccaaag ccgtacattg ggaaccggtc 3840
acacatgtaa gtgactgata taaaagagaa aaaaggcgat ttttccgcct aaaactcttt 3900
aaaacttatt aaaactctta aaacccgcct ggcctgtgca taactgtctg gccagcgcac 3960
agccgaagag ctgcaaaaag cgcctaccct tcggtcgctg cgctccctac gccccgccgc 4020
ttcgcgtcgg cctatcgcgg ccgctggccg ctcaaaaatg gctggcctac ggccaggcaa 4080
tctaccaggg cgcggacaag ccgcgccgtc gccactcgac cgccggcgcc cacatcaagg 4140
caccctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg 4200
agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt 4260
cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag 4320
tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg 4380
gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct cttccgcttc 4440
ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc 4500
aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc 4560
aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag 4620
gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc 4680
gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt 4740
tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct 4800
ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg 4860
ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct 4920
tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat 4980
tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg 5040
ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa 5100
aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt 5160
ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc 5220
tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgcattc 5280
taggtactaa aacaattcat ccagtaaaat ataatatttt attttctccc aatcaggctt 5340
gatccccagt aagtcaaaaa atagctcgac atactgttct tccccgatat cctccctgat 5400
cgaccggacg cagaaggcaa tgtcatacca cttgtccgcc ctgccgcttc tcccaagatc 5460
aataaagcca cttactttgc catctttcac aaagatgttg ctgtctccca ggtcgccgtg 5520
ggaaaagaca agttcctctt cgggcttttc cgtctttaaa aaatcataca gctcgcgcgg 5580
atctttaaat ggagtgtctt cttcccagtt ttcgcaatcc acatcggcca gatcgttatt 5640
cagtaagtaa tccaattcgg ctaagcggct gtctaagcta ttcgtatagg gacaatccga 5700
tatgtcgatg gagtgaaaga gcctgatgca ctccgcatac agctcgataa tcttttcagg 5760
gctttgttca tcttcatact cttccgagca aaggacgcca tcggcctcac tcatgagcag 5820
attgctccag ccatcatgcc gttcaaagtg caggaccttt ggaacaggca gctttccttc 5880
cagccatagc atcatgtcct tttcccgttc cacatcatag gtggtccctt tataccggct 5940
gtccgtcatt tttaaatata ggttttcatt ttctcccacc agcttatata ccttagcagg 6000
agacattcct tccgtatctt ttacgcagcg gtatttttcg atcagttttt tcaattccgg 6060
tgatattctc attttagcca tttattattt ccttcctctt ttctacagta tttaaagata 6120
ccccaagaag ctaattataa caagacgaac tccaattcac tgttccttgc attctaaaac 6180
cttaaatacc agaaaacagc tttttcaaag ttgttttcaa agttggcgta taacatagta 6240
tcgacggagc cgattttgaa accgcggtga tcacaggcag caacgctctg tcatcgttac 6300
aatcaacatg ctaccctccg cgagatcatc cgtgtttcaa acccggcagc ttagttgccg 6360
ttcttccgaa tagcatcggt aacatgagca aagtctgccg ccttacaacg gctctcccgc 6420
tgacgccgtc ccggactgat gggctgcctg tatcgagtgg tgattttgtg ccgagctgcc 6480
ggtcggggag ctgttggctg gctggtggca ggatatattg tggtgtaaac aaattgacgc 6540
ttagacaact taataacaca ttgcggacgt ttttaatgta ctgaattaac gccgaattaa 6600
ttcgggggat ctggatttta gtactggatt ttggttttag gaattagaaa ttttattgat 6660
agaagtattt tacaaataca aatacatact aagggtttct tatatgctca acacatgagc 6720
gaaaccctat aggaacccta attcccttat ctgggaacta ctcacacatt attatggaga 6780
aactcgagct tgtcgatcga cagatccggt cggcatctac tctatttctt tgccctcgga 6840
cgagtgctgg ggcgtcggtt tccactatcg gcgagtactt ctacacagcc atcggtccag 6900
acggccgcgc ttctgcgggc gatttgtgta cgcccgacag tcccggctcc ggatcggacg 6960
attgcgtcgc atcgaccctg cgcccaagct gcatcatcga aattgccgtc aaccaagctc 7020
tgatagagtt ggtcaagacc aatgcggagc atatacgccc ggagtcgtgg cgatcctgca 7080
agctccggat gcctccgctc gaagtagcgc gtctgctgct ccatacaagc caaccacggc 7140
ctccagaaga agatgttggc gacctcgtat tgggaatccc cgaacatcgc ctcgctccag 7200
tcaatgaccg ctgttatgcg gccattgtcc gtcaggacat tgttggagcc gaaatccgcg 7260
tgcacgaggt gccggacttc ggggcagtcc tcggcccaaa gcatcagctc atcgagagcc 7320
tgcgcgacgg acgcactgac ggtgtcgtcc atcacagttt gccagtgata cacatgggga 7380
tcagcaatcg cgcatatgaa atcacgccat gtagtgtatt gaccgattcc ttgcggtccg 7440
aatgggccga acccgctcgt ctggctaaga tcggccgcag cgatcgcatc catagcctcc 7500
gcgaccggtt gtagaacagc gggcagttcg gtttcaggca ggtcttgcaa cgtgacaccc 7560
tgtgcacggc gggagatgca ataggtcagg ctctcgctaa actccccaat gtcaagcact 7620
tccggaatcg ggagcgcggc cgatgcaaag tgccgataaa cataacgatc tttgtagaaa 7680
ccatcggcgc agctatttac ccgcaggaca tatccacgcc ctcctacatc gaagctgaaa 7740
gcacgagatt cttcgccctc cgagagctgc atcaggtcgg agacgctgtc gaacttttcg 7800
atcagaaact tctcgacaga cgtcgcggtg agttcaggct ttttcatatc tcattgcccc 7860
ccgggatctg cgaaagctcg agagagatag atttgtagag agagactggt gatttcagcg 7920
tgtcctctcc aaatgaaatg aacttcctta tatagaggaa ggtcttgcga aggatagtgg 7980
gattgtgcgt catcccttac gtcagtggag atatcacatc aatccacttg ctttgaagac 8040
gtggttggaa cgtcttcttt ttccacgatg ctcctcgtgg gtgggggtcc atctttggga 8100
ccactgtcgg cagaggcatc ttgaacgata gcctttcctt tatcgcaatg atggcatttg 8160
taggtgccac cttccttttc tactgtcctt ttgatgaagt gacagatagc tgggcaatgg 8220
aatccgagga ggtttcccga tattaccctt tgttgaaaag tctcaatagc cctttggtct 8280
tctgagactg tatctttgat attcttggag tagacgagag tgtcgtgctc caccatgtta 8340
tcacatcaat ccacttgctt tgaagacgtg gttggaacgt cttctttttc cacgatgctc 8400
ctcgtgggtg ggggtccatc tttgggacca ctgtcggcag aggcatcttg aacgatagcc 8460
tttcctttat cgcaatgatg gcatttgtag gtgccacctt ccttttctac tgtccttttg 8520
atgaagtgac agatagctgg gcaatggaat ccgaggaggt ttcccgatat taccctttgt 8580
tgaaaagtct caatagccct ttggtcttct gagactgtat ctttgatatt cttggagtag 8640
acgagagtgt cgtgctccac catgttggca agctgctcta gccaatacgc aaaccgcctc 8700
tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag 8760
cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt 8820
tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca 8880
caggaaacag ctatgaccat gattacgaat tcccgatcta gtaacataga tgacaccgcg 8940
cgcgataatt tatcctagtt tgcgcgctat attttgtttt ctatcgcgta ttaaatgtat 9000
aattgcggga ctctaatcat aaaaacccat ctcataaata acgtcatgca ttacatgtta 9060
attattacat gcttaacgta attcaacaga aattatatga taatcatcgc aagaccggca 9120
acaggattca atcttaagaa actttattgc caaatgtttg aacgatcggg gaaattcgag 9180
ctctaagctt aggcgccggt ggagtggcgg ccctcggcgc gctcgtactg ttccacgatg 9240
gtgtagtcct cgttgtggga ggtgatgtcc agcttgatgt cggtcttgta ggcgccgggc 9300
agctgcacgg gcttcttggc catgtaggtg gtcttgacct cggcgtcgta gtggccgccg 9360
tccttcagct tcagcctcat cttgatctcg cccttcaggg cgccgtcctc ggggtacatc 9420
cgctcggtgg aggcctccca gcccatggtc ttcttctgca ttacggggcc gtcggagggg 9480
aagttggtgc cgcgcagctt caccttgtag atgaactcgc cgtcctgcag ggaggagtcc 9540
tgggtcacgg tcaccacgcc gccgtcctcg aagttcatca cgcgctccca cttgaagccc 9600
tcggggaagg acagcttcaa gtagtcgggg atgtcggcgg ggtgcttcac gtaggccttg 9660
gagccgtact ggaactgagg ggacaggatg tcccaggcga agggcagggg gccgcccttg 9720
gtcaccttca gcttggcggt ctgggtgccc tcgtaggggc ggccctcgcc ctcgccctcg 9780
atctcgaact cgtggccgtt cacggagccc tccatgcgca ccttgaagcg catgaactcc 9840
ttgatgacgt cctcggagga ggccattgtt ataacctttc tcttcttctt aggagccatg 9900
gatccatgga gtcaaagatt caaatagagg acctaacaga actcgccgta aagactggcg 9960
aacagttcat acagagtctc ttacgactca atgacaagaa gaaaatcttc gtcaacatgg 10020
tggagcacga cacacttgtc tactccaaaa atatcaaaga tacagtctca gaagaccaaa 10080
gggcaatggt accatggctc ctaagaagaa gagaaaggtt ataacaatgg tgagcaaggg 10140
cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg 10200
ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct 10260
gaagctgatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct 10320
gggctacggc ctgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt 10380
caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg 10440
caactacaag acacgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga 10500
gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa 10560
ctacaacagc cacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccaa 10620
cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc actaccagca 10680
gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca 10740
gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt 10800
gaccgccgcc gggatcactc tcggcatgga cgaggcagct agatccacct gagtcgagtc 10860
gacggatata tgaagatgaa gatgaaatat ttggtgtgtc aaataaaaag cttgtgtgct 10920
taagtttgtg tttttttctt ggcttgttgt gttatgaatt tgtggctttt tctaatatta 10980
aatgaatgta agatcacatt ataatgaata aacaaatgtt tctataatcc attgtgaatg 11040
ttttgttgga tctcttctgc agcatataac tactgtatgt gctatggtat ggactatgga 11100
atatgattaa agataagg 11118

Claims (5)

1.一种快速鉴定水稻双向启动子的方法，其特征在于：包括如下步骤：
(1)选取已通过水稻稳定转化验证的两个双向启动子，OsBDP1和OsBDP2；
(2)构建双报告载体GFP-RFP reporter(SEQ ID NO.1)；
(3)将OsBDP1和OsBDP2扩增后分别插入双报告载体GFP-RFP reporter(SEQ ID NO.1)，构建OsBDP1 reporter和OsBDP2 reporter载体(SEQ ID NO.2，SEQ ID NO.3)；
(4)水稻原生质体的制备与转化；
(5)转化后对OsBDP1、OsBDP2的活性进行验证。
2.根据权利要求1所述的快速鉴定水稻双向启动子的方法，其特征在于：所述步骤(2)中双报告载体GFP-RFP reporter构建方法为：以pCambia1301载体为模板，扩增Ubiquitin启动子；以载体pGWB454为模板，扩增RFP编码序列；使用限制性内切酶NcoI和EcoRI将p1300LV载体线性化，用无缝连接试剂盒将上述3个片段快速连接，转化至大肠杆菌DH5α中，筛选阳性转化子。
3.根据权利要求1所述的快速鉴定水稻双向启动子的方法，其特征在于：构建负对照载体。
4.根据权利要求3所述的快速鉴定水稻双向启动子的方法，其特征在于：所述负对照载体构建方法为：使用限制性内切酶KpnI和BamHI将GFP-RFPreporter载体线性化，将2个mini35s或一段无功能序列插入线性化载体，连接后转化至大肠杆菌DH5α中，筛选阳性转化子，得到2个负对照载体mini35s reporter(SEQ ID NO.4)、promoterless reporter(SEQID NO.5)。
5.根据权利要求1所述的快速鉴定水稻双向启动子的方法，其特征在于：所述步骤(3)扩增的引物序列分别为：SEQ ID NO.6、SEQ ID NO.7、SEQ ID NO.8、SEQ ID NO.9)。
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