CN110904143A - 一种多功能抗草甘膦水稻转化载体pCDMAR-epsps及其构建方法和应用 - Google Patents

Abstract

本发明提供了一种多功能抗草甘膦水稻转化载体pCDMAR‑epsps及其构建方法和应用，其特征在于，载体pCDMAR‑epsps的大小为12774bp，序列如SEQ ID NO：1所示；构建方法包括骨架选择、含pCDMAR‑hyg骨架的感受态细胞制备及培养、酶切、连接、pCDMAR‑UbiEP中间产物的获得、带有酶切位点的T‑nos片段的获得、酶切连接。本发明的有益效果在于：本发明的一种多功能抗草甘膦水稻转化载体pCDMAR‑epsps较现有技术具有更好的安全性，而且对外源基因的稳定性和表达水平具有一定的优势，同时单插入位点中的多酶切位点还为实现同时转入2个及2个以上的多基因聚合育种的可能性。

Description

一种多功能抗草甘膦水稻转化载体pCDMAR-epsps及其构建方 法和应用

技术领域

本发明基因工程技术领域，具体涉及一种多功能抗草甘膦水稻转化载体pCDMAR-epsps及其构建方法和应用。

背景技术

杂草每年给农业生产造成的经济损失约占农作物总产值的10％～20％，因而杂草防治在农业生产中尤为重要。相比用机械除草，喷施除草剂具有省时高效的特点。草甘膦(glyphosate)是世界上应用最广泛的非选择性内吸传导性有机膦类除草剂。

但是，这种灭生性除草剂在除杂草的同时也会给农作物带来明显的伤害。目前，还未发现含有抗除草剂(草甘膦)的水稻种质资源，因此无法通过传统育种方式获得抗除草剂的水稻品种。通过基因工程技术将抗除草剂基因导入农作物，已经获得了多个抗除草剂农作物新品种，并且已经在农业生产中广泛应用。

水稻作为主要的粮食作物，对人们生活具有重要的意义，因此，抗除草剂水稻品种的开发，具有一定的需求和巨大的市场潜力。

但是，目前所构建的外源基因插入载体多以潮霉素抗性基因(hpt)作为筛选标记基因，其安全性受到一定质疑，所以开发一种更加安全、高效的多功能抗草甘膦水稻转化载体及其构建方法就显得尤为重要。

发明内容

为了弥补现有技术的不足，本发明提供了一种多功能抗草甘膦水稻转化载体pCDMAR-epsps及其构建方法和应用。本发明的载体利用抗除草剂基因，不仅可以使作物获得除草剂抗性而方便除草，本身也是筛选标记，还可以用于在田间对其它目的基因的跟踪和选择，本身的安全性也得到了广泛认同；而且本发明的转化载体对外源基因的稳定性和表达水平也进行了优化，单插入位点中的多酶切位点还为实现同时转入2个及2个以上的多基因聚合育种提供了可能性。

本发明提供了一种多功能抗草甘膦水稻转化载体pCDMAR-epsps，大小为12774bp，序列如SEQ ID NO：1所示；且所述载体pCDMAR-epsps中包括11个功能元件，具体见下表：

本发明提供了一种多功能抗草甘膦水稻转化载体pCDMAR-epsps的构建方法，包括以下步骤：

(1)骨架选择：选择pCDMAR-hyg作为骨架，且所述pCDMAR-hyg骨架大小为11724bp，序列如SEQ ID NO：14所示；

(2)含pCDMAR-hyg骨架的感受态细胞制备及培养：将pCDMAR-hyg骨架转化到感受态细胞中，并采用常规方法进行培养，获得含pCDMAR-hyg骨架的感受态细胞；

(3)酶切：采用BclI酶酶切所述pCDMAR-hyg骨架，得到片段Ⅰ、片段Ⅱ、片段Ⅲ；且所述片段Ⅰ的大小为6067bp，序列如SEQ ID NO：15所示；片段Ⅱ的大小为2975bp，序列如SEQID NO：16所示；片段Ⅲ的大小为2682bp，序列如SEQ ID NO：17所示；

(4)连接：将所述片段Ⅰ、片段Ⅱ连接得到连接产物，并通过ClaI/EcoRV酶切鉴定所述连接产物；选择ClaI/EcoRV酶切后片段大小为7346bp和1403bp的连接产物，并将其命名为中间质粒pCDMAR；所述中间质粒pCDMAR的大小为8749bp，序列如SEQ ID NO：18所示；

(5)pCDMAR-UbiEP中间产物的获得：利用SbfI/KpnI酶酶切pCUEP102质粒获得酶切产物，并回收其中大小为3783bp的片段，命名为酶切片段Ⅰ，序列如SEQ ID NO：19所示；之后利用SbfI/KpnI酶切所述中间质粒pCDMAR，回收酶切片段，命名为酶切片段Ⅱ；之后将所述酶切片段Ⅰ和酶切片段Ⅱ进行连接，获得连接产物，命名为pCDMAR-UbiEP中间产物；所述pCDMAR-UbiEP中间产物的大小为12505bp，序列如SEQ ID NO：20所示；

(6)带有酶切位点的T-nos片段的获得：设计两端加KpnI和SacI酶切位点的引物，并采用所述引物扩增pCUEP102上T-nos片段，回收大小为269bp的目的条带，且所述目的条带的序列如SEQ ID NO：8所示；

(7)酶切连接：将所述目的条带与所述pCDMAR-UbiEP中间产物通过KpnI/SacI酶进行酶切连接，得到连接产物，即为一种多功能抗草甘膦水稻转化载体pCDMAR-epsps；且所述载体pCDMAR-epsps的大小为12774bp，序列如SEQ ID NO：1所示。

作为一种优选的方案，步骤(2)中所述感受态细胞为Trans110感受态细胞。

更为优选的是，步骤(4)中所述片段Ⅰ、片段Ⅱ两条片段连接过程使用的连接酶为T4连接酶。

更为优选的是，步骤(7)中所述目的条带与所述pCDMAR-UbiEP中间产物通过KpnI/SacI酶进行酶切连接的连接过程使用的连接酶为T4连接酶。

更为优选的是，步骤(6)中所述引物包括上游引物和下游引物，且所述上游引物序列如SEQ ID NO：21所示，所述下游引物序列如SEQ ID NO：22所示。

前述的一种多功能抗草甘膦水稻转化载体pCDMAR-epsps在获得抗除草剂的水稻品种方面的应用。

前述的一种多功能抗草甘膦水稻转化载体pCDMAR-epsps在农业领域的应用。

本发明的有益效果在于：本发明提供的一种多功能抗草甘膦水稻转化载体pCDMAR-epsps及其构建方法和应用具有以下优势：

(1)本发明的一种多功能抗草甘膦水稻转化载体pCDMAR-epsps的构建方法通过改造原有双T-DNA载体，去除其选择标记基因，包括相应左右Border、启动子以及终止子等元件，得到具有双MAR序列(两个MAR序列之间的部分可以形成DNA环，使外源基因在转基因植物染色质中形成独立的区域，提高外源基因的稳定性和表达水平)、双MAR序列中间具有多克隆位点且不含有选择标记的中间载体；并将启动子、EPSPS和T-nos终止子按要求重组到这个中间质粒上，完成构建。

(2)本发明的一种多功能抗草甘膦水稻转化载体pCDMAR-epsps具有单T-DNA、抗除草剂基因两端有双MAR序列、无选择标记基因、EPSPS基因连接Ubiquitin启动子、启动子靠近Border右臂等特点。

(3)本发明的一种多功能抗草甘膦水稻转化载体pCDMAR-epsps较现有技术具有更好的安全性，而且对外源基因的稳定性和表达水平具有一定的优势，同时单插入位点中的多酶切位点还为实现同时转入2个及2个以上的多基因聚合育种的可能性。

附图说明

图1是本发明实施例1的一种多功能抗草甘膦水稻转化载体pCDMAR-epsps的构建方法的流程图；

图2是本发明实施例1的步骤(1)中使用的pCDMAR-hyg骨架的质粒结构示意图；

图3是本发明实施例1的步骤(3)中采用BclI酶酶切pCDMAR-hyg骨架获得的酶切产物电泳图；图中泳道1为1kb的Maker，泳道2为酶切产物；

图4是本发明实施例1的步骤(4)中连接产物进行ClaI/EcoRV酶切获得的酶切产物电泳图；图中泳道1为1kb的Maker，泳道2-4为3组酶切产物；

图5是本发明实施例1中获得的中间质粒pCDMAR的结构示意图；

图6是本发明实施例1的步骤(5)中使用的pCUEP102质粒的结构示意图；

图7是本发明实施例1的步骤(5)中SbfI/KpnI酶切pCUEP102质粒获得的酶切产物电泳图；图中泳道1为2kb的Maker，泳道2-7为6组酶切产物；

图8是本发明实施例1中获得的pCDMAR-UbiEP中间产物的结构示意图；

图9是本发明实施例1的步骤(6)中PCR扩增扩增pCUEP102上T-nos片段获得的PCR扩增产物电泳图；图中泳道1为100bp Maker，泳道2-3为2组PCR扩增产物；

图10是本发明实施例1的步骤(7)的流程图；

图11是本发明实施例1中获得的一种多功能抗草甘膦水稻转化载体pCDMAR-epsps的结构示意图。

具体实施方式

为使本发明实施例的目的、技术方案和优点更加清楚，对本发明实施例中的技术方案进行清楚、完整地描述，显然，所描述的实施例是本发明一部分实施例，而不是全部的实施例。基于本发明中的实施例，本领域普通技术人员在没有作出创造性劳动前提下所获得的所有其他实施例，都属于本发明保护的范围。

实施例1

1.实验材料

1.1菌株：E.coli DH 5α(根据《分子克隆》实验室制备)、E.coli Trans110(购于北京全式金生物技术有限公司，目录号：CD311).

1.2所用质粒及中间产物：pCDMAR-hyg(Constructed by our lab)、pCUEP102(Constructed by our lab)、pCDMAR、pCDMAR-UbiEP、pCDMAR-epsps。

1.3培养基：LB medium:Tryptone 10g/L,Yeast extract 5g/L,NaCl 10g/L.Adjust pH to 7.0 with 1M NaOH.For plates,add agar 15g/L.(MolecularCloning)。

1.4重要试剂：质粒小提试剂盒购自promaga公司、凝胶纯化试剂盒购自promaga公司、限制性内切酶及连接酶等购自NEB公司。

1.5重要仪器：凝胶成像仪购自Alpha Innotech公司、紫外分光光度计购自岛津公司、低温冷冻离心机购自Eppendorf公司、PCR仪购自Eppendorf公司UV1000型探照紫外仪购自UVP公司。

2.试验方法

2.1质粒构建：参考《分子克隆》中质粒DNA的制备，利用限制性内切酶消化，连接，筛选，鉴定获得重组转化子。

2.2 PCR检测：参考《分子克隆》，利用Taq酶，标准的链式反应扩增程序进行PCR反应，产物进行1％的琼脂糖电泳观察结果。

2.3大肠杆菌的转化与培养(参考分子克隆III)：连接产物冰浴30min后42℃温浴90s，热击入大肠杆菌DH5α感受态细胞，37℃温浴1h后，涂布在含有相应抗性的LB培养基上，37℃倒置培养过夜。

2.4一种多功能抗草甘膦水稻转化载体pCDMAR-epsps的构建方法，包括以下步骤，流程参见图1：

(1)骨架选择：选择pCDMAR-hyg作为骨架，且pCDMAR-hyg骨架大小为11724bp，序列如SEQ ID NO：14所示，质粒结构见示意图2；

(2)含pCDMAR-hyg骨架的感受态细胞制备及培养：将pCDMAR-hyg骨架转化到感受态细胞中，并采用常规方法进行培养，获得含pCDMAR-hyg骨架的感受态细胞；所述感受态细胞为Trans110感受态细胞；

(3)酶切：采用BclI酶酶切pCDMAR-hyg骨架，得到片段Ⅰ、片段Ⅱ、片段Ⅲ(酶切产物电泳图参见图3)；且片段Ⅰ的大小为6067bp，序列如SEQ ID NO：15所示；片段Ⅱ的大小为2975bp，序列如SEQ ID NO：16所示；片段Ⅲ的大小为2682bp，序列如SEQ ID NO：17所示；

(4)连接：将片段Ⅰ、片段Ⅱ通过T4连接酶连接得到连接产物，并通过ClaI/EcoRV酶切鉴定连接产物(酶切产物电泳图参见图4)；选择ClaI/EcoRV酶切后的片段大小为7346bp和1403bp的连接产物，并将其命名为中间质粒pCDMAR；中间质粒pCDMAR的大小为8749bp，序列如SEQ ID NO：18所示，其结构示意图参见图5；

(5)pCDMAR-UbiEP中间产物的获得：利用SbfI/KpnI酶切pCUEP102质粒(pCUEP102质粒结构参见图6)，并回收酶切产物(酶切产物电泳图参见图7)中大小为3783bp的片段，命名为酶切片段Ⅰ，序列如SEQ ID NO：19所示；利用SbfI/KpnI酶切中间质粒pCDMAR，回收酶切片段，命名为酶切片段Ⅱ；之后将酶切片段Ⅰ和酶切片段Ⅱ进行连接，获得连接产物，命名为pCDMAR-UbiEP中间产物；pCDMAR-UbiEP中间产物大小为12505bp，序列如SEQ ID NO：20所示，结构示意图参见图8；

(6)带有酶切位点的T-nos片段的获得：设计两端加KpnI和SacI酶切位点的引物，并采用引物扩增pCUEP102上T-nos片段(PCR扩增产物电泳图参见图9)，回收大小为269bp的目的条带，且目的条带的序列如SEQ ID NO：8所示；引物包括上游引物和下游引物，且上游引物序列如SEQ ID NO：21所示，下游引物序列如SEQ ID NO：22所示；

(7)酶切连接(具体流程参见图10)：将目的条带与pCDMAR-UbiEP中间产物通过KpnI/SacI酶进行酶切，之后通过T4连接酶连接，得到连接产物即为一种多功能抗草甘膦水稻转化载体pCDMAR-epsps；且载体pCDMAR-epsps的大小为12774bp，序列如SEQ ID NO：1所示，其结构示意图参见图11。

应当理解，以上所描述的具体实施例仅用于解释本发明，并不用于限定本发明。由本发明的精神所引伸出的显而易见的变化或变动仍处于本发明的保护范围之中。

序列表

<110> 黑龙江省农业科学院耕作栽培研究所

<120> 一种多功能抗草甘膦水稻转化载体pCDMAR-epsps及其构建方法和应用

<130> 2019-08-30

<160> 22

<170> SIPOSequenceListing 1.0

<210> 1

<211> 12774

<212> DNA

<213> Escherichia coli

<400> 1

catgccaacc acagggttcc cctcgggatc aaagtacttt gatccaaccc ctccgctgct 60

atagtgcagt cggcttctga cgttcagtgc agccgtcttc tgaaaacgac atgtcgcaca 120

agtcctaagt tacgcgacag gctgccgccc tgcccttttc ctggcgtttt cttgtcgcgt 180

gttttagtcg cataaagtag aatacttgcg actagaaccg gagacattac gccatgaaca 240

agagcgccgc cgctggcctg ctgggctatg cccgcgtcag caccgacgac caggacttga 300

ccaaccaacg ggccgaactg cacgcggccg gctgcaccaa gctgttttcc gagaagatca 360

ccggcaccag gcgcgaccgc ccggagctgg ccaggatgct tgaccaccta cgccctggcg 420

acgttgtgac agtgaccagg ctagaccgcc tggcccgcag cacccgcgac ctactggaca 480

ttgccgagcg catccaggag gccggcgcgg gcctgcgtag cctggcagag ccgtgggccg 540

acaccaccac gccggccggc cgcatggtgt tgaccgtgtt cgccggcatt gccgagttcg 600

agcgttccct aatcatcgac cgcacccgga gcgggcgcga ggccgccaag gcccgaggcg 660

tgaagtttgg cccccgccct accctcaccc cggcacagat cgcgcacgcc cgcgagctga 720

tcgaccagga aggccgcacc gtgaaagagg cggctgcact gcttggcgtg catcgctcga 780

ccctgtaccg cgcacttgag cgcagcgagg aagtgacgcc caccgaggcc aggcggcgcg 840

gtgccttccg tgaggacgca ttgaccgagg ccgacgccct ggcggccgcc gagaatgaac 900

gccaagagga acaagcatga aaccgcacca ggacggccag gacgaaccgt ttttcattac 960

cgaagagatc gaggcggaga tgatcgcggc cgggtacgtg ttcgagccgc ccgcgcacgt 1020

ctcaaccgtg cggctgcatg aaatcctggc cggtttgtct gatgccaagc tggcggcctg 1080

gccggccagc ttggccgctg aagaaaccga gcgccgccgt ctaaaaaggt gatgtgtatt 1140

tgagtaaaac agcttgcgtc atgcggtcgc tgcgtatatg atgcgatgag taaataaaca 1200

aatacgcaag gggaacgcat gaaggttatc gctgtactta accagaaagg cgggtcaggc 1260

aagacgacca tcgcaaccca tctagcccgc gccctgcaac tcgccggggc cgatgttctg 1320

ttagtcgatt ccgatcccca gggcagtgcc cgcgattggg cggccgtgcg ggaagatcaa 1380

ccgctaaccg ttgtcggcat cgaccgcccg acgattgacc gcgacgtgaa ggccatcggc 1440

cggcgcgact tcgtagtgat cgacggagcg ccccaggcgg cggacttggc tgtgtccgcg 1500

atcaaggcag ccgacttcgt gctgattccg gtgcagccaa gcccttacga catatgggcc 1560

accgccgacc tggtggagct ggttaagcag cgcattgagg tcacggatgg aaggctacaa 1620

gcggcctttg tcgtgtcgcg ggcgatcaaa ggcacgcgca tcggcggtga ggttgccgag 1680

gcgctggccg ggtacgagct gcccattctt gagtcccgta tcacgcagcg cgtgagctac 1740

ccaggcactg ccgccgccgg cacaaccgtt cttgaatcag aacccgaggg cgacgctgcc 1800

cgcgaggtcc aggcgctggc cgctgaaatt aaatcaaaac tcatttgagt taatgaggta 1860

aagagaaaat gagcaaaagc acaaacacgc taagtgccgg ccgtccgagc gcacgcagca 1920

gcaaggctgc aacgttggcc agcctggcag acacgccagc catgaagcgg gtcaactttc 1980

agttgccggc ggaggatcac accaagctga agatgtacgc ggtacgccaa ggcaagacca 2040

ttaccgagct gctatctgaa tacatcgcgc agctaccaga gtaaatgagc aaatgaataa 2100

atgagtagat gaattttagc ggctaaagga ggcggcatgg aaaatcaaga acaaccaggc 2160

accgacgccg tggaatgccc catgtgtgga ggaacgggcg gttggccagg cgtaagcggc 2220

tgggttgtct gccggccctg caatggcact ggaaccccca agcccgagga atcggcgtga 2280

cggtcgcaaa ccatccggcc cggtacaaat cggcgcggcg ctgggtgatg acctggtgga 2340

gaagttgaag gccgcgcagg ccgcccagcg gcaacgcatc gaggcagaag cacgccccgg 2400

tgaatcgtgg caagcggccg ctgatcgaat ccgcaaagaa tcccggcaac cgccggcagc 2460

cggtgcgccg tcgattagga agccgcccaa gggcgacgag caaccagatt ttttcgttcc 2520

gatgctctat gacgtgggca cccgcgatag tcgcagcatc atggacgtgg ccgttttccg 2580

tctgtcgaag cgtgaccgac gagctggcga ggtgatccgc tacgagcttc cagacgggca 2640

cgtagaggtt tccgcagggc cggccggcat ggccagtgtg tgggattacg acctggtact 2700

gatggcggtt tcccatctaa ccgaatccat gaaccgatac cgggaaggga agggagacaa 2760

gcccggccgc gtgttccgtc cacacgttgc ggacgtactc aagttctgcc ggcgagccga 2820

tggcggaaag cagaaagacg acctggtaga aacctgcatt cggttaaaca ccacgcacgt 2880

tgccatgcag cgtacgaaga aggccaagaa cggccgcctg gtgacggtat ccgagggtga 2940

agccttgatt agccgctaca agatcgtaaa gagcgaaacc gggcggccgg agtacatcga 3000

gatcgagcta gctgattgga tgtaccgcga gatcacagaa ggcaagaacc cggacgtgct 3060

gacggttcac cccgattact ttttgatcga tcccggcatc ggccgttttc tctaccgcct 3120

ggcacgccgc gccgcaggca aggcagaagc cagatggttg ttcaagacga tctacgaacg 3180

cagtggcagc gccggagagt tcaagaagtt ctgtttcacc gtgcgcaagc tgatcgggtc 3240

aaatgacctg ccggagtacg atttgaagga ggaggcgggg caggctggcc cgatcctagt 3300

catgcgctac cgcaacctga tcgagggcga agcatccgcc ggttcctaat gtacggagca 3360

gatgctaggg caaattgccc tagcagggga aaaaggtcga aaaggtctct ttcctgtgga 3420

tagcacgtac attgggaacc caaagccgta cattgggaac cggaacccgt acattgggaa 3480

cccaaagccg tacattggga accggtcaca catgtaagtg actgatataa aagagaaaaa 3540

aggcgatttt tccgcctaaa actctttaaa acttattaaa actcttaaaa cccgcctggc 3600

ctgtgcataa ctgtctggcc agcgcacagc cgaagagctg caaaaagcgc ctacccttcg 3660

gtcgctgcgc tccctacgcc ccgccgcttc gcgtcggcct atcgcggccg ctggccgctc 3720

aaaaatggct ggcctacggc caggcaatct accagggcgc ggacaagccg cgccgtcgcc 3780

actcgaccgc cggcgcccac atcaaggcac cctgcctcgc gcgtttcggt gatgacggtg 3840

aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg 3900

ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca 3960

tgacccagtc acgtagcgat agcggagtgt atactggctt aactatgcgg catcagagca 4020

gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa 4080

ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg 4140

gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg 4200

ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa 4260

ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 4320

acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 4380

tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 4440

ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc 4500

ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg 4560

ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc 4620

actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga 4680

gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc 4740

tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac 4800

caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 4860

atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 4920

acgttaaggg attttggtca tgcattctag gtactaaaac aattcatcca gtaaaatata 4980

atattttatt ttctcccaat caggcttgat ccccagtaag tcaaaaaata gctcgacata 5040

ctgttcttcc ccgatatcct ccctgatcga ccggacgcag aaggcaatgt cataccactt 5100

gtccgccctg ccgcttctcc caagatcaat aaagccactt actttgccat ctttcacaaa 5160

gatgttgctg tctcccaggt cgccgtggga aaagacaagt tcctcttcgg gcttttccgt 5220

ctttaaaaaa tcatacagct cgcgcggatc tttaaatgga gtgtcttctt cccagttttc 5280

gcaatccaca tcggccagat cgttattcag taagtaatcc aattcggcta agcggctgtc 5340

taagctattc gtatagggac aatccgatat gtcgatggag tgaaagagcc tgatgcactc 5400

cgcatacagc tcgataatct tttcagggct ttgttcatct tcatactctt ccgagcaaag 5460

gacgccatcg gcctcactca tgagcagatt gctccagcca tcatgccgtt caaagtgcag 5520

gacctttgga acaggcagct ttccttccag ccatagcatc atgtcctttt cccgttccac 5580

atcataggtg gtccctttat accggctgtc cgtcattttt aaatataggt tttcattttc 5640

tcccaccagc ttatatacct tagcaggaga cattccttcc gtatctttta cgcagcggta 5700

tttttcgatc agttttttca attccggtga tattctcatt ttagccattt attatttcct 5760

tcctcttttc tacagtattt aaagataccc caagaagcta attataacaa gacgaactcc 5820

aattcactgt tccttgcatt ctaaaacctt aaataccaga aaacagcttt ttcaaagttg 5880

ttttcaaagt tggcgtataa catagtatcg acggagccga ttttgaaacc ggtgatcaca 5940

ggcagcaacg ctctgtcatc gttacaatca acatgctacc ctccgcgaga tcatccgtgt 6000

ttcaaacccg gcagcttagt tgccgttctt ccgaatagca tcggtaacat gagcaaagtc 6060

tgccgcctta caacggctct cccgctgacg ccgtcccgga ctgatgggct gcctgtatcg 6120

agtggtgatt ttgtgccgag ctgccggtcg gggagctgtt ggctggctgg tggcaggata 6180

tattgtggtg taaacaaatt gacgcttaga caacttaata acacattgcg gacgttttta 6240

atgtactgaa ttaacgccga attaattcgg gggatctgga ttttagtact ggattttggt 6300

tttaggaatt agaaatttta ttgatagaag tattttacaa atacaaatac atactaaggg 6360

tttcttatat gctcaacaca tgagcgaaac cctataggaa ccctaattcc cttatctggg 6420

aactactcac acattattat ggagaaactc gacggtatcg ataagcttga tccatgcctc 6480

acatgttaat gtactaccaa tggaggcttc catgcctcac atgttcatgt acacatttat 6540

gattaggaaa ctttttaata tattttatag atttcttatc catcatataa aaatacataa 6600

ttaatcatac gattttgaga tacatattct gacgtatcaa attctaatta aattttaaaa 6660

tattttagtg acgtatcaaa ttctaattaa attttaaaat attttagaga cgtattttcg 6720

taacaattta aaatgtatat tatagatcac attcataggt cattttataa tttaaaatat 6780

tatggagatg catcttcgtt tatttttacg gagatatatt ttcgtaattt atcataatag 6840

aattgttcat gctatatttt gtttatgttt gctcagatga agatttaaac cttacaagca 6900

atgtgcaaaa aatgacgtac ataaatttag atggtccaaa aatgttataa ataaaagatc 6960

aagaagtgtc aaaaaaagtc aaaaacaacg atagagtagt ataatgtcaa aataaaataa 7020

aatccatgac actactacta ttatatatta atgcactaat gtgtatgtct aactacatcg 7080

cctctgcctc ctctgtcagt tatgtctcgt aggccatcaa tcccccgtcc tccgacgttg 7140

tctccggtac atcaatgtcc catgtgccta cgtcatgatg gcatttagga catgtctcac 7200

atcagccaga tcagcaagat acatttgtca atgtctatct acgcaatctc cacaatgcga 7260

cgacatatag gcaagacatc ctcaacataa tttagttgtg catgcttctc ctctagtatc 7320

tcccgatgag ttgatcgaat taattcctgc agcccttggc aagctgctct agccaatacg 7380

caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc 7440

cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc 7500

accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata 7560

acaatttcac acaggaaaca gctatgacat gattacgaat tcgagctcga tctagtaaca 7620

tagatgacac cgcgcgcgat aatttatcct agtttgcgcg ctatattttg ttttctatcg 7680

cgtattaaat gtataattgc gggactctaa tcataaaaac ccatctcata aataacgtca 7740

tgcattacat gttaattatt acatgcttaa cgtaattcaa cagaaattat atgataatca 7800

tcgcaagacc ggcaacagga ttcaatctta agaaacttta ttgccaaatg tttgaacgat 7860

cggggaaatt cgagctcggt accgggcccc ccctcgaggt cgacggtatc gataagcttg 7920

atatcgaatt cctgcagctc actcttttaa aagctcagtt cagttcctga cgaaagtgct 7980

tagaacgtcg aagtagttgg ggaaggtctt gcgggtgcaa ccagggtccc tgatcgtcac 8040

gggcacgtcg gcgcaggcag cgagggagaa ggccatggcc atcctgtgat catcgtaggt 8100

gtcgattgcc gtgatgttca gcttctccgg tggggtgatg atgcagtagt caggaccttc 8160

ttcaaccgat gctcccagct ttgttagctc ggtccgaatt gcaaccatcc tttcggtttc 8220

ctttactctc caggaagcca catctctgat agcagttgga ccatcagcga agagtgcaac 8280

aacggcaagg gtcatggcaa catcaggcat tttgttcatg ttgacatcaa cagctttcag 8340

gtgtttcttc ccataaggct cacgtggtgg accagttacg gttacactgg tgtcagtcca 8400

tgtaaccttt gctcccatca tctcaagtac ctcagcaaat ttgacatcac cctgcaaact 8460

ggtcgtacca caaccttgaa ctgtcacagt gcctccagtg attgcagcac cagccaagaa 8520

atagctcgcg cttgaggcat caccttcaac ataggcattt ccaggagatt tgtacttctg 8580

ccctccctta atatagaatc tgtcccaact atcagaatgc tctgccttca caccaaaacg 8640

ctccatcaat ctcaatgtca tttcaacgta aggaatggag attagtttgt caatgatttc 8700

gatctccaca tccccaaggg ccaaaggagc agccatcagc aaggcactca agtactgact 8760

gctgatggaa ccagagagct taaccttgcc accaggaagt cctccaattc ccttgacacg 8820

aacaggtggg cattcagtgc caaggaaaca gtcgacatcc gcaccaagtt gtttcaaccc 8880

gacaaccaag tcaccaatcg gtctctccct cattcgtggc actccatcaa gcacataagt 8940

tgcatttcca ccagcagcag tcacggctgc tgtcaatagt cgcattgcag ttccagcgtt 9000

ccccaagaag agttgcactt cctctttcgc atccttctca acaggaaact tgccaccaca 9060

gccaacgact acagctcttt ttgcaacttt atctgcttcc acagagagcc cgagggcttt 9120

cagggcctca agcatgtagt gaacatcctc actgttcagc aagttgtcca ccactgttgt 9180

gccctcggag agggcggaga ggaggaggat cctgttggag agcgacttgg accctggcag 9240

ctgaaccgcc ccggagatct ccctgatggg ctggagcacg atctcctccg ccttcgccgc 9300

cggcgctgcc accgacgacg acgacgcgga cgccaccacc accgcctccc gccgcccccg 9360

cgcccgcacc cgcacccgca tccccccgcg cgccgcggcg ggcagccgca gctgcttccg 9420

cgacgagaac gccgccgacg ccgccacggc ctggtccagg gacaccgccg ccgcagccgc 9480

ggcgttggac gccatggtcg ccgccattct agagcggccg ctctagaact agtggatccg 9540

aagtaacacc aaacaacagg gtgagcatcg acaaaagaaa cagtaccaag caaataaata 9600

gcgtatgaag gcagggctaa aaaaatccac atatagctgc tgcatatgcc atcatccaag 9660

tatatcaaga tcaaaataat tataaaacat acttgtttat tataatagat aggtactcaa 9720

ggttagagca tatgaataga tgctgcatat gccatcatgt atatgcatca gtaaaaccca 9780

catcaacatg tatacctatc ctagatcgat atttccatcc atcttaaact cgtaactatg 9840

aagatgtatg acacacacat acagttccaa aattaataaa tacaccaggt agtttgaaac 9900

agtattctac tccgatctag aacgaatgaa cgaccgccca accacaccac atcatcacaa 9960

ccaagcgaac aaaaagcatc tctgtatatg catcagtaaa acccgcatca acatgtatac 10020

ctatcctaga tcgatatttc catccatcat cttcaattcg taactatgaa tatgtatggc 10080

acacacatac agatccaaaa ttaataaatc caccaggtag tttgaaacag aattctactc 10140

cgatctagaa cgaccgccca accagaccac atcatcacaa ccaagacaaa aaaaagcatg 10200

aaaagatgac ccgacaaaca agtgcacggc atatattgaa ataaaggaaa agggcaaacc 10260

aaaccctatg caacgaaaca aaaaaaatca tgaaatcgat cccgtctgcg gaacggctag 10320

agccatccca ggattcccca aagagaaaca ctggcaagtt agcaatcaga acgtgtctga 10380

cgtacaggtc gcatccgtgt acgaacgcta gcagcacgga tctaacacaa acacggatct 10440

aacacaaaca tgaacagaag tagaactacc gggccctaac catggaccgg aacgccgatc 10500

tagagaaggt agagaggggg ggggggggag gacgagcggc gtaccttgaa gcggaggtgc 10560

cgacgggtgg atttggggga gatctggttg tgtgtgtgtg cgctccgaac aacacgaggt 10620

tggggaaaga gggtgtggag ggggtgtcta tttattacgg cgggcgagga agggaaagcg 10680

aaggagcggt gggaaaggaa tcccccgtag ctgccgtgcc gtgagaggag gaggaggccg 10740

cctgccgtgc cggctcacgt ctgccgctcc gccacgcaat ttctggatgc cgacagcgga 10800

gcaagtccaa cggtggagcg gaactctcga gaggggtcca gaggcagcga cagagatgcc 10860

gtgccgtctg cttcgcttgg cccgacgcga cgctgctggt tcgctggttg gtgtccgtta 10920

gactcgtcga cggcgtttaa caggctggca ttatctactc gaaacaagaa aaatgtttcc 10980

ttagtttttt taatttctta aagggtattt gtttaatttt tagtcacttt attttattct 11040

attttatatc taaattatta aataaaaaaa ctaaaataga gttttagttt tcttaattta 11100

gaggctaaaa tagaataaaa tagatgtact aaaaaaatta gtctataaaa accattaacc 11160

ctaaacccta aatggatgta ctaataaaat ggatgaagta ttatataggt gaagctattt 11220

gcaaaaaaaa aggagaacac atgcacacta aaaagataaa actgtagagt cctgttgtca 11280

aaatactcaa ttgtccttta gaccatgtct aactgttcat ttatatgatt ctctaaaaca 11340

ctgatattat tgtagtacta tagattatat tattcgtaga gtaaagttta aatatatgta 11400

taaagataga taaactgcac ttcaacaagc ttggcactgg ccgtcgtttt acaacgtcgt 11460

gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc 11520

agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg 11580

aatggcgaat gctagagcag cttgagcttg gatcagattg tcgtttcccg ccttcagttt 11640

ggggatcctc tagagtcgac ctgcaggcat gcaagcttgg cactggccgt cgttttacaa 11700

cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct 11760

ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc 11820

agcctgaatg gcgaatgcta gagcagcttg agcttggatc agattgtcgt ttcccgcctt 11880

cagtttaatt cgatccatgc ctcacatgtt aatgtactac caatggaggg ctgtacacat 11940

ttatgattac gaaatttttt aatatatttt atagatttct tatgcatcat acaaaaatac 12000

ataattattc gtaacatttt ggagatacat attcagatgc atcaaattct aattaaacgt 12060

taaaatattt tggagacgta tcttcgtaac aatttaaaac ctatactata catcacattc 12120

gaaggtcatt ttataattta aaatattatg gagatgcatc ttcgtttatg tttgctcaga 12180

tgaagattta aaccttacaa acaatatgta aaaaatgacg tacataaatt cagatagtcc 12240

aaaagtgtca tatataaata aagatcaata agtgtcaaaa aaagtcaaga acaacgatag 12300

agtagcataa tgtcaaaata aaataaaatc catgacacta ctactattat atattaatgc 12360

actaatgtgt atgtctaact acatcgtctc tgcctcctct gtcagttatg tctcgtaagc 12420

catcaatccc ccgtcctccg gcgttgtctc cggtatatca atgtccccat gtgcctacgt 12480

catgatggca tctaggacat gtctcacatc agacacatta ggaaagatac atttgccaat 12540

gtatatctgc gcaatctcca caatgcaacg acatataggc aagacatcct caacataatt 12600

tagttgtgca tgcttctcct ctagtatctc ccgatgagtt gatcaagctt atcgaaacta 12660

tcagtgtttg acaggatata ttggcgggta aacctaagag aaaagagcgt ttattagaat 12720

aacggatatt taaaagggcg tgaaaaggtt tatccgttcg tccatttgta tgtg 12774

<210> 2

<211> 26

<212> DNA

<213> Escherichia coli

<400> 2

gtaaacctaa gagaaaagag cgttta 26

<210> 3

<211> 26

<212> DNA

<213> Escherichia coli

<400> 3

tggcaggata tattgtggtg taaaca 26

<210> 4

<211> 747

<212> DNA

<213> Escherichia coli

<400> 4

ccatgcctca catgttaatg tactaccaat ggagggctgt acacatttat gattacgaaa 60

ttttttaata tattttatag atttcttatg catcatacaa aaatacataa ttattcgtaa 120

cattttggag atacatattc agatgcatca aattctaatt aaacgttaaa atattttgga 180

gacgtatctt cgtaacaatt taaaacctat actatacatc acattcgaag gtcattttat 240

aatttaaaat attatggaga tgcatcttcg tttatgtttg ctcagatgaa gatttaaacc 300

ttacaaacaa tatgtaaaaa atgacgtaca taaattcaga tagtccaaaa gtgtcatata 360

taaataaaga tcaataagtg tcaaaaaaag tcaagaacaa cgatagagta gcataatgtc 420

aaaataaaat aaaatccatg acactactac tattatatat taatgcacta atgtgtatgt 480

ctaactacat cgtctctgcc tcctctgtca gttatgtctc gtaagccatc aatcccccgt 540

cctccggcgt tgtctccggt atatcaatgt ccccatgtgc ctacgtcatg atggcatcta 600

ggacatgtct cacatcagac acattaggaa agatacattt gccaatgtat atctgcgcaa 660

tctccacaat gcaacgacat ataggcaaga catcctcaac ataatttagt tgtgcatgct 720

tctcctctag tatctcccga tgagttg 747

<210> 5

<211> 862

<212> DNA

<213> Escherichia coli

<400> 5

ccatgcctca catgttaatg tactaccaat ggaggcttcc atgcctcaca tgttcatgta 60

cacatttatg attaggaaac tttttaatat attttataga tttcttatcc atcatataaa 120

aatacataat taatcatacg attttgagat acatattctg acgtatcaaa ttctaattaa 180

attttaaaat attttagtga cgtatcaaat tctaattaaa ttttaaaata ttttagagac 240

gtattttcgt aacaatttaa aatgtatatt atagatcaca ttcataggtc attttataat 300

ttaaaatatt atggagatgc atcttcgttt atttttacgg agatatattt tcgtaattta 360

tcataataga attgttcatg ctatattttg tttatgtttg ctcagatgaa gatttaaacc 420

ttacaagcaa tgtgcaaaaa atgacgtaca taaatttaga tggtccaaaa atgttataaa 480

taaaagatca agaagtgtca aaaaaagtca aaaacaacga tagagtagta taatgtcaaa 540

ataaaataaa atccatgaca ctactactat tatatattaa tgcactaatg tgtatgtcta 600

actacatcgc ctctgcctcc tctgtcagtt atgtctcgta ggccatcaat cccccgtcct 660

ccgacgttgt ctccggtaca tcaatgtccc atgtgcctac gtcatgatgg catttaggac 720

atgtctcaca tcagccagat cagcaagata catttgtcaa tgtctatcta cgcaatctcc 780

acaatgcgac gacatatagg caagacatcc tcaacataat ttagttgtgc atgcttctcc 840

tctagtatct cccgatgagt tg 862

<210> 6

<211> 1911

<212> DNA

<213> Escherichia coli

<400> 6

tctagaacta gtggatccga agtaacacca aacaacaggg tgagcatcga caaaagaaac 60

agtaccaagc aaataaatag cgtatgaagg cagggctaaa aaaatccaca tatagctgct 120

gcatatgcca tcatccaagt atatcaagat caaaataatt ataaaacata cttgtttatt 180

ataatagata ggtactcaag gttagagcat atgaatagat gctgcatatg ccatcatgta 240

tatgcatcag taaaacccac atcaacatgt atacctatcc tagatcgata tttccatcca 300

tcttaaactc gtaactatga agatgtatga cacacacata cagttccaaa attaataaat 360

acaccaggta gtttgaaaca gtattctact ccgatctaga acgaatgaac gaccgcccaa 420

ccacaccaca tcatcacaac caagcgaaca aaaagcatct ctgtatatgc atcagtaaaa 480

cccgcatcaa catgtatacc tatcctagat cgatatttcc atccatcatc ttcaattcgt 540

aactatgaat atgtatggca cacacataca gatccaaaat taataaatcc accaggtagt 600

ttgaaacaga attctactcc gatctagaac gaccgcccaa ccagaccaca tcatcacaac 660

caagacaaaa aaaagcatga aaagatgacc cgacaaacaa gtgcacggca tatattgaaa 720

taaaggaaaa gggcaaacca aaccctatgc aacgaaacaa aaaaaatcat gaaatcgatc 780

ccgtctgcgg aacggctaga gccatcccag gattccccaa agagaaacac tggcaagtta 840

gcaatcagaa cgtgtctgac gtacaggtcg catccgtgta cgaacgctag cagcacggat 900

ctaacacaaa cacggatcta acacaaacat gaacagaagt agaactaccg ggccctaacc 960

atggaccgga acgccgatct agagaaggta gagagggggg gggggggagg acgagcggcg 1020

taccttgaag cggaggtgcc gacgggtgga tttgggggag atctggttgt gtgtgtgtgc 1080

gctccgaaca acacgaggtt ggggaaagag ggtgtggagg gggtgtctat ttattacggc 1140

gggcgaggaa gggaaagcga aggagcggtg ggaaaggaat cccccgtagc tgccgtgccg 1200

tgagaggagg aggaggccgc ctgccgtgcc ggctcacgtc tgccgctccg ccacgcaatt 1260

tctggatgcc gacagcggag caagtccaac ggtggagcgg aactctcgag aggggtccag 1320

aggcagcgac agagatgccg tgccgtctgc ttcgcttggc ccgacgcgac gctgctggtt 1380

cgctggttgg tgtccgttag actcgtcgac ggcgtttaac aggctggcat tatctactcg 1440

aaacaagaaa aatgtttcct tagttttttt aatttcttaa agggtatttg tttaattttt 1500

agtcacttta ttttattcta ttttatatct aaattattaa ataaaaaaac taaaatagag 1560

ttttagtttt cttaatttag aggctaaaat agaataaaat agatgtacta aaaaaattag 1620

tctataaaaa ccattaaccc taaaccctaa atggatgtac taataaaatg gatgaagtat 1680

tatataggtg aagctatttg caaaaaaaaa ggagaacaca tgcacactaa aaagataaaa 1740

ctgtagagtc ctgttgtcaa aatactcaat tgtcctttag accatgtcta actgttcatt 1800

tatatgattc tctaaaacac tgatattatt gtagtactat agattatatt attcgtagag 1860

taaagtttaa atatatgtat aaagatagat aaactgcact tcaacaagct t 1911

<210> 7

<211> 1644

<212> DNA

<213> Escherichia coli

<400> 7

ggtaccgggc cccccctcga ggtcgacggt atcgataagc ttgatatcga attcctgcag 60

ctcactcttt taaaagctca gttcagttcc tgacgaaagt gcttagaacg tcgaagtagt 120

tggggaaggt cttgcgggtg caaccagggt ccctgatcgt cacgggcacg tcggcgcagg 180

cagcgaggga gaaggccatg gccatcctgt gatcatcgta ggtgtcgatt gccgtgatgt 240

tcagcttctc cggtggggtg atgatgcagt agtcaggacc ttcttcaacc gatgctccca 300

gctttgttag ctcggtccga attgcaacca tcctttcggt ttcctttact ctccaggaag 360

ccacatctct gatagcagtt ggaccatcag cgaagagtgc aacaacggca agggtcatgg 420

caacatcagg cattttgttc atgttgacat caacagcttt caggtgtttc ttcccataag 480

gctcacgtgg tggaccagtt acggttacac tggtgtcagt ccatgtaacc tttgctccca 540

tcatctcaag tacctcagca aatttgacat caccctgcaa actggtcgta ccacaacctt 600

gaactgtcac agtgcctcca gtgattgcag caccagccaa gaaatagctc gcgcttgagg 660

catcaccttc aacataggca tttccaggag atttgtactt ctgccctccc ttaatataga 720

atctgtccca actatcagaa tgctctgcct tcacaccaaa acgctccatc aatctcaatg 780

tcatttcaac gtaaggaatg gagattagtt tgtcaatgat ttcgatctcc acatccccaa 840

gggccaaagg agcagccatc agcaaggcac tcaagtactg actgctgatg gaaccagaga 900

gcttaacctt gccaccagga agtcctccaa ttcccttgac acgaacaggt gggcattcag 960

tgccaaggaa acagtcgaca tccgcaccaa gttgtttcaa cccgacaacc aagtcaccaa 1020

tcggtctctc cctcattcgt ggcactccat caagcacata agttgcattt ccaccagcag 1080

cagtcacggc tgctgtcaat agtcgcattg cagttccagc gttccccaag aagagttgca 1140

cttcctcttt cgcatccttc tcaacaggaa acttgccacc acagccaacg actacagctc 1200

tttttgcaac tttatctgct tccacagaga gcccgagggc tttcagggcc tcaagcatgt 1260

agtgaacatc ctcactgttc agcaagttgt ccaccactgt tgtgccctcg gagagggcgg 1320

agaggaggag gatcctgttg gagagcgact tggaccctgg cagctgaacc gccccggaga 1380

tctccctgat gggctggagc acgatctcct ccgccttcgc cgccggcgct gccaccgacg 1440

acgacgacgc ggacgccacc accaccgcct cccgccgccc ccgcgcccgc acccgcaccc 1500

gcatcccccc gcgcgccgcg gcgggcagcc gcagctgctt ccgcgacgag aacgccgccg 1560

acgccgccac ggcctggtcc agggacaccg ccgccgcagc cgcggcgttg gacgccatgg 1620

tcgccgccat tctagagcgg ccgc 1644

<210> 8

<211> 269

<212> DNA

<213> Escherichia coli

<400> 8

gatctagtaa catagatgac accgcgcgcg ataatttatc ctagtttgcg cgctatattt 60

tgttttctat cgcgtattaa atgtataatt gcgggactct aatcataaaa acccatctca 120

taaataacgt catgcattac atgttaatta ttacatgctt aacgtaattc aacagaaatt 180

atatgataat catcgcaaga ccggcaacag gattcaatct taagaaactt tattgccaaa 240

tgtttgaacg atcggggaaa ttcgagctc 269

<210> 9

<211> 792

<212> DNA

<213> Escherichia coli

<400> 9

aaacaattca tccagtaaaa tataatattt tattttctcc caatcaggct tgatccccag 60

taagtcaaaa aatagctcga catactgttc ttccccgata tcctccctga tcgaccggac 120

gcagaaggca atgtcatacc acttgtccgc cctgccgctt ctcccaagat caataaagcc 180

acttactttg ccatctttca caaagatgtt gctgtctccc aggtcgccgt gggaaaagac 240

aagttcctct tcgggctttt ccgtctttaa aaaatcatac agctcgcgcg gatctttaaa 300

tggagtgtct tcttcccagt tttcgcaatc cacatcggcc agatcgttat tcagtaagta 360

atccaattcg gctaagcggc tgtctaagct attcgtatag ggacaatccg atatgtcgat 420

ggagtgaaag agcctgatgc actccgcata cagctcgata atcttttcag ggctttgttc 480

atcttcatac tcttccgagc aaaggacgcc atcggcctca ctcatgagca gattgctcca 540

gccatcatgc cgttcaaagt gcaggacctt tggaacaggc agctttcctt ccagccatag 600

catcatgtcc ttttcccgtt ccacatcata ggtggtccct ttataccggc tgtccgtcat 660

ttttaaatat aggttttcat tttctcccac cagcttatat accttagcag gagacattcc 720

ttccgtatct tttacgcagc ggtatttttc gatcagtttt ttcaattccg gtgatattct 780

cattttagcc at 792

<210> 10

<211> 281

<212> DNA

<213> Escherichia coli

<400> 10

gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct 60

ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg 120

tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct 180

gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac 240

tggcagcagc cactggtaac aggattagca gagcgaggta t 281

<210> 11

<211> 261

<212> DNA

<213> Escherichia coli

<400> 11

cggagtgtat actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat 60

atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgctcttcc 120

gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct 180

cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg 240

tgagcaaaag gccagcaaaa g 261

<210> 12

<211> 1001

<212> DNA

<213> Escherichia coli

<400> 12

gttttccgtc tgtcgaagcg tgaccgacga gctggcgagg tgatccgcta cgagcttcca 60

gacgggcacg tagaggtttc cgcagggccg gccggcatgg ccagtgtgtg ggattacgac 120

ctggtactga tggcggtttc ccatctaacc gaatccatga accgataccg ggaagggaag 180

ggagacaagc ccggccgcgt gttccgtcca cacgttgcgg acgtactcaa gttctgccgg 240

cgagccgatg gcggaaagca gaaagacgac ctggtagaaa cctgcattcg gttaaacacc 300

acgcacgttg ccatgcagcg tacgaagaag gccaagaacg gccgcctggt gacggtatcc 360

gagggtgaag ccttgattag ccgctacaag atcgtaaaga gcgaaaccgg gcggccggag 420

tacatcgaga tcgagctagc tgattggatg taccgcgaga tcacagaagg caagaacccg 480

gacgtgctga cggttcaccc cgattacttt ttgatcgatc ccggcatcgg ccgttttctc 540

taccgcctgg cacgccgcgc cgcaggcaag gcagaagcca gatggttgtt caagacgatc 600

tacgaacgca gtggcagcgc cggagagttc aagaagttct gtttcaccgt gcgcaagctg 660

atcgggtcaa atgacctgcc ggagtacgat ttgaaggagg aggcggggca ggctggcccg 720

atcctagtca tgcgctaccg caacctgatc gagggcgaag catccgccgg ttcctaatgt 780

acggagcaga tgctagggca aattgcccta gcaggggaaa aaggtcgaaa aggtctcttt 840

cctgtggata gcacgtacat tgggaaccca aagccgtaca ttgggaaccg gaacccgtac 900

attgggaacc caaagccgta cattgggaac cggtcacaca tgtaagtgac tgatataaaa 960

gagaaaaaag gcgatttttc cgcctaaaac tctttaaaac t 1001

<210> 13

<211> 1001

<212> DNA

<213> Escherichia coli

<400> 13

atgatcgcgg ccgggtacgt gttcgagccg cccgcgcacg tctcaaccgt gcggctgcat 60

gaaatcctgg ccggtttgtc tgatgccaag ctggcggcct ggccggccag cttggccgct 120

gaagaaaccg agcgccgccg tctaaaaagg tgatgtgtat ttgagtaaaa cagcttgcgt 180

catgcggtcg ctgcgtatat gatgcgatga gtaaataaac aaatacgcaa ggggaacgca 240

tgaaggttat cgctgtactt aaccagaaag gcgggtcagg caagacgacc atcgcaaccc 300

atctagcccg cgccctgcaa ctcgccgggg ccgatgttct gttagtcgat tccgatcccc 360

agggcagtgc ccgcgattgg gcggccgtgc gggaagatca accgctaacc gttgtcggca 420

tcgaccgccc gacgattgac cgcgacgtga aggccatcgg ccggcgcgac ttcgtagtga 480

tcgacggagc gccccaggcg gcggacttgg ctgtgtccgc gatcaaggca gccgacttcg 540

tgctgattcc ggtgcagcca agcccttacg acatatgggc caccgccgac ctggtggagc 600

tggttaagca gcgcattgag gtcacggatg gaaggctaca agcggccttt gtcgtgtcgc 660

gggcgatcaa aggcacgcgc atcggcggtg aggttgccga ggcgctggcc gggtacgagc 720

tgcccattct tgagtcccgt atcacgcagc gcgtgagcta cccaggcact gccgccgccg 780

gcacaaccgt tcttgaatca gaacccgagg gcgacgctgc ccgcgaggtc caggcgctgg 840

ccgctgaaat taaatcaaaa ctcatttgag ttaatgaggt aaagagaaaa tgagcaaaag 900

cacaaacacg ctaagtgccg gccgtccgag cgcacgcagc agcaaggctg caacgttggc 960

cagcctggca gacacgccag ccatgaagcg ggtcaacttt c 1001

<210> 14

<211> 11724

<212> DNA

<213> Escherichia coli

<400> 14

catgccaacc acagggttcc cctcgggatc aaagtacttt gatccaaccc ctccgctgct 60

atagtgcagt cggcttctga cgttcagtgc agccgtcttc tgaaaacgac atgtcgcaca 120

agtcctaagt tacgcgacag gctgccgccc tgcccttttc ctggcgtttt cttgtcgcgt 180

gttttagtcg cataaagtag aatacttgcg actagaaccg gagacattac gccatgaaca 240

agagcgccgc cgctggcctg ctgggctatg cccgcgtcag caccgacgac caggacttga 300

ccaaccaacg ggccgaactg cacgcggccg gctgcaccaa gctgttttcc gagaagatca 360

ccggcaccag gcgcgaccgc ccggagctgg ccaggatgct tgaccaccta cgccctggcg 420

acgttgtgac agtgaccagg ctagaccgcc tggcccgcag cacccgcgac ctactggaca 480

ttgccgagcg catccaggag gccggcgcgg gcctgcgtag cctggcagag ccgtgggccg 540

acaccaccac gccggccggc cgcatggtgt tgaccgtgtt cgccggcatt gccgagttcg 600

agcgttccct aatcatcgac cgcacccgga gcgggcgcga ggccgccaag gcccgaggcg 660

tgaagtttgg cccccgccct accctcaccc cggcacagat cgcgcacgcc cgcgagctga 720

tcgaccagga aggccgcacc gtgaaagagg cggctgcact gcttggcgtg catcgctcga 780

ccctgtaccg cgcacttgag cgcagcgagg aagtgacgcc caccgaggcc aggcggcgcg 840

gtgccttccg tgaggacgca ttgaccgagg ccgacgccct ggcggccgcc gagaatgaac 900

gccaagagga acaagcatga aaccgcacca ggacggccag gacgaaccgt ttttcattac 960

cgaagagatc gaggcggaga tgatcgcggc cgggtacgtg ttcgagccgc ccgcgcacgt 1020

ctcaaccgtg cggctgcatg aaatcctggc cggtttgtct gatgccaagc tggcggcctg 1080

gccggccagc ttggccgctg aagaaaccga gcgccgccgt ctaaaaaggt gatgtgtatt 1140

tgagtaaaac agcttgcgtc atgcggtcgc tgcgtatatg atgcgatgag taaataaaca 1200

aatacgcaag gggaacgcat gaaggttatc gctgtactta accagaaagg cgggtcaggc 1260

aagacgacca tcgcaaccca tctagcccgc gccctgcaac tcgccggggc cgatgttctg 1320

ttagtcgatt ccgatcccca gggcagtgcc cgcgattggg cggccgtgcg ggaagatcaa 1380

ccgctaaccg ttgtcggcat cgaccgcccg acgattgacc gcgacgtgaa ggccatcggc 1440

cggcgcgact tcgtagtgat cgacggagcg ccccaggcgg cggacttggc tgtgtccgcg 1500

atcaaggcag ccgacttcgt gctgattccg gtgcagccaa gcccttacga catatgggcc 1560

accgccgacc tggtggagct ggttaagcag cgcattgagg tcacggatgg aaggctacaa 1620

gcggcctttg tcgtgtcgcg ggcgatcaaa ggcacgcgca tcggcggtga ggttgccgag 1680

gcgctggccg ggtacgagct gcccattctt gagtcccgta tcacgcagcg cgtgagctac 1740

ccaggcactg ccgccgccgg cacaaccgtt cttgaatcag aacccgaggg cgacgctgcc 1800

cgcgaggtcc aggcgctggc cgctgaaatt aaatcaaaac tcatttgagt taatgaggta 1860

aagagaaaat gagcaaaagc acaaacacgc taagtgccgg ccgtccgagc gcacgcagca 1920

gcaaggctgc aacgttggcc agcctggcag acacgccagc catgaagcgg gtcaactttc 1980

agttgccggc ggaggatcac accaagctga agatgtacgc ggtacgccaa ggcaagacca 2040

ttaccgagct gctatctgaa tacatcgcgc agctaccaga gtaaatgagc aaatgaataa 2100

atgagtagat gaattttagc ggctaaagga ggcggcatgg aaaatcaaga acaaccaggc 2160

accgacgccg tggaatgccc catgtgtgga ggaacgggcg gttggccagg cgtaagcggc 2220

tgggttgtct gccggccctg caatggcact ggaaccccca agcccgagga atcggcgtga 2280

cggtcgcaaa ccatccggcc cggtacaaat cggcgcggcg ctgggtgatg acctggtgga 2340

gaagttgaag gccgcgcagg ccgcccagcg gcaacgcatc gaggcagaag cacgccccgg 2400

tgaatcgtgg caagcggccg ctgatcgaat ccgcaaagaa tcccggcaac cgccggcagc 2460

cggtgcgccg tcgattagga agccgcccaa gggcgacgag caaccagatt ttttcgttcc 2520

gatgctctat gacgtgggca cccgcgatag tcgcagcatc atggacgtgg ccgttttccg 2580

tctgtcgaag cgtgaccgac gagctggcga ggtgatccgc tacgagcttc cagacgggca 2640

cgtagaggtt tccgcagggc cggccggcat ggccagtgtg tgggattacg acctggtact 2700

gatggcggtt tcccatctaa ccgaatccat gaaccgatac cgggaaggga agggagacaa 2760

gcccggccgc gtgttccgtc cacacgttgc ggacgtactc aagttctgcc ggcgagccga 2820

tggcggaaag cagaaagacg acctggtaga aacctgcatt cggttaaaca ccacgcacgt 2880

tgccatgcag cgtacgaaga aggccaagaa cggccgcctg gtgacggtat ccgagggtga 2940

agccttgatt agccgctaca agatcgtaaa gagcgaaacc gggcggccgg agtacatcga 3000

gatcgagcta gctgattgga tgtaccgcga gatcacagaa ggcaagaacc cggacgtgct 3060

gacggttcac cccgattact ttttgatcga tcccggcatc ggccgttttc tctaccgcct 3120

ggcacgccgc gccgcaggca aggcagaagc cagatggttg ttcaagacga tctacgaacg 3180

cagtggcagc gccggagagt tcaagaagtt ctgtttcacc gtgcgcaagc tgatcgggtc 3240

aaatgacctg ccggagtacg atttgaagga ggaggcgggg caggctggcc cgatcctagt 3300

catgcgctac cgcaacctga tcgagggcga agcatccgcc ggttcctaat gtacggagca 3360

gatgctaggg caaattgccc tagcagggga aaaaggtcga aaaggtctct ttcctgtgga 3420

tagcacgtac attgggaacc caaagccgta cattgggaac cggaacccgt acattgggaa 3480

cccaaagccg tacattggga accggtcaca catgtaagtg actgatataa aagagaaaaa 3540

aggcgatttt tccgcctaaa actctttaaa acttattaaa actcttaaaa cccgcctggc 3600

ctgtgcataa ctgtctggcc agcgcacagc cgaagagctg caaaaagcgc ctacccttcg 3660

gtcgctgcgc tccctacgcc ccgccgcttc gcgtcggcct atcgcggccg ctggccgctc 3720

aaaaatggct ggcctacggc caggcaatct accagggcgc ggacaagccg cgccgtcgcc 3780

actcgaccgc cggcgcccac atcaaggcac cctgcctcgc gcgtttcggt gatgacggtg 3840

aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg 3900

ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca 3960

tgacccagtc acgtagcgat agcggagtgt atactggctt aactatgcgg catcagagca 4020

gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa 4080

ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg 4140

gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg 4200

ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa 4260

ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 4320

acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 4380

tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 4440

ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc 4500

ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg 4560

ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc 4620

actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga 4680

gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc 4740

tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac 4800

caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 4860

atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 4920

acgttaaggg attttggtca tgcattctag gtactaaaac aattcatcca gtaaaatata 4980

atattttatt ttctcccaat caggcttgat ccccagtaag tcaaaaaata gctcgacata 5040

ctgttcttcc ccgatatcct ccctgatcga ccggacgcag aaggcaatgt cataccactt 5100

gtccgccctg ccgcttctcc caagatcaat aaagccactt actttgccat ctttcacaaa 5160

gatgttgctg tctcccaggt cgccgtggga aaagacaagt tcctcttcgg gcttttccgt 5220

ctttaaaaaa tcatacagct cgcgcggatc tttaaatgga gtgtcttctt cccagttttc 5280

gcaatccaca tcggccagat cgttattcag taagtaatcc aattcggcta agcggctgtc 5340

taagctattc gtatagggac aatccgatat gtcgatggag tgaaagagcc tgatgcactc 5400

cgcatacagc tcgataatct tttcagggct ttgttcatct tcatactctt ccgagcaaag 5460

gacgccatcg gcctcactca tgagcagatt gctccagcca tcatgccgtt caaagtgcag 5520

gacctttgga acaggcagct ttccttccag ccatagcatc atgtcctttt cccgttccac 5580

atcataggtg gtccctttat accggctgtc cgtcattttt aaatataggt tttcattttc 5640

tcccaccagc ttatatacct tagcaggaga cattccttcc gtatctttta cgcagcggta 5700

tttttcgatc agttttttca attccggtga tattctcatt ttagccattt attatttcct 5760

tcctcttttc tacagtattt aaagataccc caagaagcta attataacaa gacgaactcc 5820

aattcactgt tccttgcatt ctaaaacctt aaataccaga aaacagcttt ttcaaagttg 5880

ttttcaaagt tggcgtataa catagtatcg acggagccga ttttgaaacc ggtgatcaca 5940

ggcagcaacg ctctgtcatc gttacaatca acatgctacc ctccgcgaga tcatccgtgt 6000

ttcaaacccg gcagcttagt tgccgttctt ccgaatagca tcggtaacat gagcaaagtc 6060

tgccgcctta caacggctct cccgctgacg ccgtcccgga ctgatgggct gcctgtatcg 6120

agtggtgatt ttgtgccgag ctgccggtcg gggagctgtt ggctggctgg tggcaggata 6180

tattgtggtg taaacaaatt gacgcttaga caacttaata acacattgcg gacgttttta 6240

atgtactgaa ttaacgccga attaattcgg gggatctgga ttttagtact ggattttggt 6300

tttaggaatt agaaatttta ttgatagaag tattttacaa atacaaatac atactaaggg 6360

tttcttatat gctcaacaca tgagcgaaac cctataggaa ccctaattcc cttatctggg 6420

aactactcac acattattat ggagaaactc gagcttgtcg atcgacagat ccggtcggca 6480

tctactctat ttctttgccc tcggacgagt gctggggcgt cggtttccac tatcggcgag 6540

tacttctaca cagccatcgg tccagacggc cgcgcttctg cgggcgattt gtgtacgccc 6600

gacagtcccg gctccggatc ggacgattgc gtcgcatcga ccctgcgccc aagctgcatc 6660

atcgaaattg ccgtcaacca agctctgata gagttggtca agaccaatgc ggagcatata 6720

cgcccggagt cgtggcgatc ctgcaagctc cggatgcctc cgctcgaagt agcgcgtctg 6780

ctgctccata caagccaacc acggcctcca gaagaagatg ttggcgacct cgtattggga 6840

atccccgaac atcgcctcgc tccagtcaat gaccgctgtt atgcggccat tgtccgtcag 6900

gacattgttg gagccgaaat ccgcgtgcac gaggtgccgg acttcggggc agtcctcggc 6960

ccaaagcatc agctcatcga gagcctgcgc gacggacgca ctgacggtgt cgtccatcac 7020

agtttgccag tgatacacat ggggatcagc aatcgcgcat atgaaatcac gccatgtagt 7080

gtattgaccg attccttgcg gtccgaatgg gccgaacccg ctcgtctggc taagatcggc 7140

cgcagcgatc gcatccatag cctccgcgac cggttgtaga acagcgggca gttcggtttc 7200

aggcaggtct tgcaacgtga caccctgtgc acggcgggag atgcaatagg tcaggctctc 7260

gctaaactcc ccaatgtcaa gcacttccgg aatcgggagc gcggccgatg caaagtgccg 7320

ataaacataa cgatctttgt agaaaccatc ggcgcagcta tttacccgca ggacatatcc 7380

acgccctcct acatcgaagc tgaaagcacg agattcttcg ccctccgaga gctgcatcag 7440

gtcggagacg ctgtcgaact tttcgatcag aaacttctcg acagacgtcg cggtgagttc 7500

aggctttttc atatctcatt gcccccccgg atctgcgaaa gctcgagaga gatagatttg 7560

tagagagaga ctggtgattt cagcgtgtcc tctccaaatg aaatgaactt ccttatatag 7620

aggaaggtct tgcgaaggat agtgggattg tgcgtcatcc cttacgtcag tggagatatc 7680

acatcaatcc acttgctttg aagacgtggt tggaacgtct tctttttcca cgatgctcct 7740

cgtgggtggg ggtccatctt tgggaccact gtcggcagag gcatcttgaa cgatagcctt 7800

tcctttatcg caatgatggc atttgtaggt gccaccttcc ttttctactg tccttttgat 7860

gaagtgacag atagctgggc aatggaatcc gaggaggttt cccgatatta ccctttgttg 7920

aaaagtctca atagcccttt ggtcttctga gactgtatct ttgatattct tggagtagac 7980

gagagtgtcg tgctccacca tgttatcaca tcaatccact tgctttgaag acgtggttgg 8040

aacgtcttct ttttccacga tgctcctcgt gggtgggggt ccatctttgg gaccactgtc 8100

ggcagaggca tcttgaacga tagcctttcc tttatcgcaa tgatggcatt tgtaggtgcc 8160

accttccttt tctactgtcc ttttgatgaa gtgacagata gctgggcaat ggaatccgag 8220

gaggtttccc gatattaccc tttgttgaaa agtctcaata gccctttggt cttctgagac 8280

tgtatctttg atattcttgg agtagacgag agtgtcgtgc tccaccatgt tggcaagctg 8340

ctctagccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg 8400

cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag 8460

ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga 8520

attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgaatttggc 8580

actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg 8640

ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg 8700

cccttcccaa cagttgcgca gcctgaatgg cgaatgctag agcagcttga gcttggatca 8760

gattgtcgtt tcccgccttc agtttaaact atcagtgttt gacaggatat attggcgggt 8820

aaacctaaga gaaaagagcg tttattagaa taacggatat ttaaaagggc gtgaaaaggt 8880

ttatccgttc gtccatttgt atgtgggtga tcacaggcag caacgctctg tcatcgttac 8940

aatcaacatg ctaccctccg cgagatcatc cgtgtttcaa acccggcagc ttagttgccg 9000

ttcttccgaa tagcatcggt aacatgagca aagtctgccg ccttacaacg gctctcccgc 9060

tgacgccgtc ccggactgat gggctgcctg tatcgagtgg tgattttgtg ccgagctgcc 9120

ggtcggggag ctgttggctg gctggtggca ggatatattg tggtgtaaac aaattgacgc 9180

ttagacaact taataacaca ttgcggacgt ttttaatgta ctgaattaac gccgaattaa 9240

ttcgggggat ctggatttta gtactggatt ttggttttag gaattagaaa ttttattgat 9300

agaagtattt tacaaataca aatacatact aagggtttct tatatgctca acacatgagc 9360

gaaaccctat aggaacccta attcccttat ctgggaacta ctcacacatt attatggaga 9420

aactcgacgg tatcgataag cttgatccat gcctcacatg ttaatgtact accaatggag 9480

gcttccatgc ctcacatgtt catgtacaca tttatgatta ggaaactttt taatatattt 9540

tatagatttc ttatccatca tataaaaata cataattaat catacgattt tgagatacat 9600

attctgacgt atcaaattct aattaaattt taaaatattt tagtgacgta tcaaattcta 9660

attaaatttt aaaatatttt agagacgtat tttcgtaaca atttaaaatg tatattatag 9720

atcacattca taggtcattt tataatttaa aatattatgg agatgcatct tcgtttattt 9780

ttacggagat atattttcgt aatttatcat aatagaattg ttcatgctat attttgttta 9840

tgtttgctca gatgaagatt taaaccttac aagcaatgtg caaaaaatga cgtacataaa 9900

tttagatggt ccaaaaatgt tataaataaa agatcaagaa gtgtcaaaaa aagtcaaaaa 9960

caacgataga gtagtataat gtcaaaataa aataaaatcc atgacactac tactattata 10020

tattaatgca ctaatgtgta tgtctaacta catcgcctct gcctcctctg tcagttatgt 10080

ctcgtaggcc atcaatcccc cgtcctccga cgttgtctcc ggtacatcaa tgtcccatgt 10140

gcctacgtca tgatggcatt taggacatgt ctcacatcag ccagatcagc aagatacatt 10200

tgtcaatgtc tatctacgca atctccacaa tgcgacgaca tataggcaag acatcctcaa 10260

cataatttag ttgtgcatgc ttctcctcta gtatctcccg atgagttgat cgaattaatt 10320

cctgcagccc ttggcaagct gctctagcca atacgcaaac cgcctctccc cgcgcgttgg 10380

ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc 10440

aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt 10500

ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat 10560

gacatgatta cgaattcgag ctcggtaccc ggggatcctc tagagtcgac ctgcaggcat 10620

gcaagcttgg cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc 10680

caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc 10740

cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatgcta gagcagcttg 10800

agcttggatc agattgtcgt ttcccgcctt cagtttaatt cgatccatgc ctcacatgtt 10860

aatgtactac caatggaggg ctgtacacat ttatgattac gaaatttttt aatatatttt 10920

atagatttct tatgcatcat acaaaaatac ataattattc gtaacatttt ggagatacat 10980

attcagatgc atcaaattct aattaaacgt taaaatattt tggagacgta tcttcgtaac 11040

aatttaaaac ctatactata catcacattc gaaggtcatt ttataattta aaatattatg 11100

gagatgcatc ttcgtttatg tttgctcaga tgaagattta aaccttacaa acaatatgta 11160

aaaaatgacg tacataaatt cagatagtcc aaaagtgtca tatataaata aagatcaata 11220

agtgtcaaaa aaagtcaaga acaacgatag agtagcataa tgtcaaaata aaataaaatc 11280

catgacacta ctactattat atattaatgc actaatgtgt atgtctaact acatcgtctc 11340

tgcctcctct gtcagttatg tctcgtaagc catcaatccc ccgtcctccg gcgttgtctc 11400

cggtatatca atgtccccat gtgcctacgt catgatggca tctaggacat gtctcacatc 11460

agacacatta ggaaagatac atttgccaat gtatatctgc gcaatctcca caatgcaacg 11520

acatataggc aagacatcct caacataatt tagttgtgca tgcttctcct ctagtatctc 11580

ccgatgagtt gatcaagctt atcgaaacta tcagtgtttg acaggatata ttggcgggta 11640

aacctaagag aaaagagcgt ttattagaat aacggatatt taaaagggcg tgaaaaggtt 11700

tatccgttcg tccatttgta tgtg 11724

<210> 15

<211> 6067

<212> DNA

<213> 人工序列(Artificial sequence)

<400> 15

gatcaagctt atcgaaacta tcagtgtttg acaggatata ttggcgggta aacctaagag 60

aaaagagcgt ttattagaat aacggatatt taaaagggcg tgaaaaggtt tatccgttcg 120

tccatttgta tgtgcatgcc aaccacaggg ttcccctcgg gatcaaagta ctttgatcca 180

acccctccgc tgctatagtg cagtcggctt ctgacgttca gtgcagccgt cttctgaaaa 240

cgacatgtcg cacaagtcct aagttacgcg acaggctgcc gccctgccct tttcctggcg 300

ttttcttgtc gcgtgtttta gtcgcataaa gtagaatact tgcgactaga accggagaca 360

ttacgccatg aacaagagcg ccgccgctgg cctgctgggc tatgcccgcg tcagcaccga 420

cgaccaggac ttgaccaacc aacgggccga actgcacgcg gccggctgca ccaagctgtt 480

ttccgagaag atcaccggca ccaggcgcga ccgcccggag ctggccagga tgcttgacca 540

cctacgccct ggcgacgttg tgacagtgac caggctagac cgcctggccc gcagcacccg 600

cgacctactg gacattgccg agcgcatcca ggaggccggc gcgggcctgc gtagcctggc 660

agagccgtgg gccgacacca ccacgccggc cggccgcatg gtgttgaccg tgttcgccgg 720

cattgccgag ttcgagcgtt ccctaatcat cgaccgcacc cggagcgggc gcgaggccgc 780

caaggcccga ggcgtgaagt ttggcccccg ccctaccctc accccggcac agatcgcgca 840

cgcccgcgag ctgatcgacc aggaaggccg caccgtgaaa gaggcggctg cactgcttgg 900

cgtgcatcgc tcgaccctgt accgcgcact tgagcgcagc gaggaagtga cgcccaccga 960

ggccaggcgg cgcggtgcct tccgtgagga cgcattgacc gaggccgacg ccctggcggc 1020

cgccgagaat gaacgccaag aggaacaagc atgaaaccgc accaggacgg ccaggacgaa 1080

ccgtttttca ttaccgaaga gatcgaggcg gagatgatcg cggccgggta cgtgttcgag 1140

ccgcccgcgc acgtctcaac cgtgcggctg catgaaatcc tggccggttt gtctgatgcc 1200

aagctggcgg cctggccggc cagcttggcc gctgaagaaa ccgagcgccg ccgtctaaaa 1260

aggtgatgtg tatttgagta aaacagcttg cgtcatgcgg tcgctgcgta tatgatgcga 1320

tgagtaaata aacaaatacg caaggggaac gcatgaaggt tatcgctgta cttaaccaga 1380

aaggcgggtc aggcaagacg accatcgcaa cccatctagc ccgcgccctg caactcgccg 1440

gggccgatgt tctgttagtc gattccgatc cccagggcag tgcccgcgat tgggcggccg 1500

tgcgggaaga tcaaccgcta accgttgtcg gcatcgaccg cccgacgatt gaccgcgacg 1560

tgaaggccat cggccggcgc gacttcgtag tgatcgacgg agcgccccag gcggcggact 1620

tggctgtgtc cgcgatcaag gcagccgact tcgtgctgat tccggtgcag ccaagccctt 1680

acgacatatg ggccaccgcc gacctggtgg agctggttaa gcagcgcatt gaggtcacgg 1740

atggaaggct acaagcggcc tttgtcgtgt cgcgggcgat caaaggcacg cgcatcggcg 1800

gtgaggttgc cgaggcgctg gccgggtacg agctgcccat tcttgagtcc cgtatcacgc 1860

agcgcgtgag ctacccaggc actgccgccg ccggcacaac cgttcttgaa tcagaacccg 1920

agggcgacgc tgcccgcgag gtccaggcgc tggccgctga aattaaatca aaactcattt 1980

gagttaatga ggtaaagaga aaatgagcaa aagcacaaac acgctaagtg ccggccgtcc 2040

gagcgcacgc agcagcaagg ctgcaacgtt ggccagcctg gcagacacgc cagccatgaa 2100

gcgggtcaac tttcagttgc cggcggagga tcacaccaag ctgaagatgt acgcggtacg 2160

ccaaggcaag accattaccg agctgctatc tgaatacatc gcgcagctac cagagtaaat 2220

gagcaaatga ataaatgagt agatgaattt tagcggctaa aggaggcggc atggaaaatc 2280

aagaacaacc aggcaccgac gccgtggaat gccccatgtg tggaggaacg ggcggttggc 2340

caggcgtaag cggctgggtt gtctgccggc cctgcaatgg cactggaacc cccaagcccg 2400

aggaatcggc gtgacggtcg caaaccatcc ggcccggtac aaatcggcgc ggcgctgggt 2460

gatgacctgg tggagaagtt gaaggccgcg caggccgccc agcggcaacg catcgaggca 2520

gaagcacgcc ccggtgaatc gtggcaagcg gccgctgatc gaatccgcaa agaatcccgg 2580

caaccgccgg cagccggtgc gccgtcgatt aggaagccgc ccaagggcga cgagcaacca 2640

gattttttcg ttccgatgct ctatgacgtg ggcacccgcg atagtcgcag catcatggac 2700

gtggccgttt tccgtctgtc gaagcgtgac cgacgagctg gcgaggtgat ccgctacgag 2760

cttccagacg ggcacgtaga ggtttccgca gggccggccg gcatggccag tgtgtgggat 2820

tacgacctgg tactgatggc ggtttcccat ctaaccgaat ccatgaaccg ataccgggaa 2880

gggaagggag acaagcccgg ccgcgtgttc cgtccacacg ttgcggacgt actcaagttc 2940

tgccggcgag ccgatggcgg aaagcagaaa gacgacctgg tagaaacctg cattcggtta 3000

aacaccacgc acgttgccat gcagcgtacg aagaaggcca agaacggccg cctggtgacg 3060

gtatccgagg gtgaagcctt gattagccgc tacaagatcg taaagagcga aaccgggcgg 3120

ccggagtaca tcgagatcga gctagctgat tggatgtacc gcgagatcac agaaggcaag 3180

aacccggacg tgctgacggt tcaccccgat tactttttga tcgatcccgg catcggccgt 3240

tttctctacc gcctggcacg ccgcgccgca ggcaaggcag aagccagatg gttgttcaag 3300

acgatctacg aacgcagtgg cagcgccgga gagttcaaga agttctgttt caccgtgcgc 3360

aagctgatcg ggtcaaatga cctgccggag tacgatttga aggaggaggc ggggcaggct 3420

ggcccgatcc tagtcatgcg ctaccgcaac ctgatcgagg gcgaagcatc cgccggttcc 3480

taatgtacgg agcagatgct agggcaaatt gccctagcag gggaaaaagg tcgaaaaggt 3540

ctctttcctg tggatagcac gtacattggg aacccaaagc cgtacattgg gaaccggaac 3600

ccgtacattg ggaacccaaa gccgtacatt gggaaccggt cacacatgta agtgactgat 3660

ataaaagaga aaaaaggcga tttttccgcc taaaactctt taaaacttat taaaactctt 3720

aaaacccgcc tggcctgtgc ataactgtct ggccagcgca cagccgaaga gctgcaaaaa 3780

gcgcctaccc ttcggtcgct gcgctcccta cgccccgccg cttcgcgtcg gcctatcgcg 3840

gccgctggcc gctcaaaaat ggctggccta cggccaggca atctaccagg gcgcggacaa 3900

gccgcgccgt cgccactcga ccgccggcgc ccacatcaag gcaccctgcc tcgcgcgttt 3960

cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct 4020

gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg 4080

tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg gcttaactat 4140

gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga 4200

tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac tgactcgctg 4260

cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta 4320

tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc 4380

aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag 4440

catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac 4500

caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc 4560

ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt 4620

aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc 4680

gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga 4740

cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta 4800

ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta 4860

tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga 4920

tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg 4980

cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag 5040

tggaacgaaa actcacgtta agggattttg gtcatgcatt ctaggtacta aaacaattca 5100

tccagtaaaa tataatattt tattttctcc caatcaggct tgatccccag taagtcaaaa 5160

aatagctcga catactgttc ttccccgata tcctccctga tcgaccggac gcagaaggca 5220

atgtcatacc acttgtccgc cctgccgctt ctcccaagat caataaagcc acttactttg 5280

ccatctttca caaagatgtt gctgtctccc aggtcgccgt gggaaaagac aagttcctct 5340

tcgggctttt ccgtctttaa aaaatcatac agctcgcgcg gatctttaaa tggagtgtct 5400

tcttcccagt tttcgcaatc cacatcggcc agatcgttat tcagtaagta atccaattcg 5460

gctaagcggc tgtctaagct attcgtatag ggacaatccg atatgtcgat ggagtgaaag 5520

agcctgatgc actccgcata cagctcgata atcttttcag ggctttgttc atcttcatac 5580

tcttccgagc aaaggacgcc atcggcctca ctcatgagca gattgctcca gccatcatgc 5640

cgttcaaagt gcaggacctt tggaacaggc agctttcctt ccagccatag catcatgtcc 5700

ttttcccgtt ccacatcata ggtggtccct ttataccggc tgtccgtcat ttttaaatat 5760

aggttttcat tttctcccac cagcttatat accttagcag gagacattcc ttccgtatct 5820

tttacgcagc ggtatttttc gatcagtttt ttcaattccg gtgatattct cattttagcc 5880

atttattatt tccttcctct tttctacagt atttaaagat accccaagaa gctaattata 5940

acaagacgaa ctccaattca ctgttccttg cattctaaaa ccttaaatac cagaaaacag 6000

ctttttcaaa gttgttttca aagttggcgt ataacatagt atcgacggag ccgattttga 6060

aaccggt 6067

<210> 16

<211> 2975

<212> DNA

<213> 人工序列(Artificial sequence)

<400> 16

gatcacaggc agcaacgctc tgtcatcgtt acaatcaaca tgctaccctc cgcgagatca 60

tccgtgtttc aaacccggca gcttagttgc cgttcttccg aatagcatcg gtaacatgag 120

caaagtctgc cgccttacaa cggctctccc gctgacgccg tcccggactg atgggctgcc 180

tgtatcgagt ggtgattttg tgccgagctg ccggtcgggg agctgttggc tggctggtgg 240

caggatatat tgtggtgtaa acaaattgac gcttagacaa cttaataaca cattgcggac 300

gtttttaatg tactgaatta acgccgaatt aattcggggg atctggattt tagtactgga 360

ttttggtttt aggaattaga aattttattg atagaagtat tttacaaata caaatacata 420

ctaagggttt cttatatgct caacacatga gcgaaaccct ataggaaccc taattccctt 480

atctgggaac tactcacaca ttattatgga gaaactcgag cttgtcgatc gacagatccg 540

gtcggcatct actctatttc tttgccctcg gacgagtgct ggggcgtcgg tttccactat 600

cggcgagtac ttctacacag ccatcggtcc agacggccgc gcttctgcgg gcgatttgtg 660

tacgcccgac agtcccggct ccggatcgga cgattgcgtc gcatcgaccc tgcgcccaag 720

ctgcatcatc gaaattgccg tcaaccaagc tctgatagag ttggtcaaga ccaatgcgga 780

gcatatacgc ccggagtcgt ggcgatcctg caagctccgg atgcctccgc tcgaagtagc 840

gcgtctgctg ctccatacaa gccaaccacg gcctccagaa gaagatgttg gcgacctcgt 900

attgggaatc cccgaacatc gcctcgctcc agtcaatgac cgctgttatg cggccattgt 960

ccgtcaggac attgttggag ccgaaatccg cgtgcacgag gtgccggact tcggggcagt 1020

cctcggccca aagcatcagc tcatcgagag cctgcgcgac ggacgcactg acggtgtcgt 1080

ccatcacagt ttgccagtga tacacatggg gatcagcaat cgcgcatatg aaatcacgcc 1140

atgtagtgta ttgaccgatt ccttgcggtc cgaatgggcc gaacccgctc gtctggctaa 1200

gatcggccgc agcgatcgca tccatagcct ccgcgaccgg ttgtagaaca gcgggcagtt 1260

cggtttcagg caggtcttgc aacgtgacac cctgtgcacg gcgggagatg caataggtca 1320

ggctctcgct aaactcccca atgtcaagca cttccggaat cgggagcgcg gccgatgcaa 1380

agtgccgata aacataacga tctttgtaga aaccatcggc gcagctattt acccgcagga 1440

catatccacg ccctcctaca tcgaagctga aagcacgaga ttcttcgccc tccgagagct 1500

gcatcaggtc ggagacgctg tcgaactttt cgatcagaaa cttctcgaca gacgtcgcgg 1560

tgagttcagg ctttttcata tctcattgcc cccccggatc tgcgaaagct cgagagagat 1620

agatttgtag agagagactg gtgatttcag cgtgtcctct ccaaatgaaa tgaacttcct 1680

tatatagagg aaggtcttgc gaaggatagt gggattgtgc gtcatccctt acgtcagtgg 1740

agatatcaca tcaatccact tgctttgaag acgtggttgg aacgtcttct ttttccacga 1800

tgctcctcgt gggtgggggt ccatctttgg gaccactgtc ggcagaggca tcttgaacga 1860

tagcctttcc tttatcgcaa tgatggcatt tgtaggtgcc accttccttt tctactgtcc 1920

ttttgatgaa gtgacagata gctgggcaat ggaatccgag gaggtttccc gatattaccc 1980

tttgttgaaa agtctcaata gccctttggt cttctgagac tgtatctttg atattcttgg 2040

agtagacgag agtgtcgtgc tccaccatgt tatcacatca atccacttgc tttgaagacg 2100

tggttggaac gtcttctttt tccacgatgc tcctcgtggg tgggggtcca tctttgggac 2160

cactgtcggc agaggcatct tgaacgatag cctttccttt atcgcaatga tggcatttgt 2220

aggtgccacc ttccttttct actgtccttt tgatgaagtg acagatagct gggcaatgga 2280

atccgaggag gtttcccgat attacccttt gttgaaaagt ctcaatagcc ctttggtctt 2340

ctgagactgt atctttgata ttcttggagt agacgagagt gtcgtgctcc accatgttgg 2400

caagctgctc tagccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg 2460

cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt 2520

gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt 2580

gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacga 2640

atttggcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac 2700

ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca 2760

ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgctagagc agcttgagct 2820

tggatcagat tgtcgtttcc cgccttcagt ttaaactatc agtgtttgac aggatatatt 2880

ggcgggtaaa cctaagagaa aagagcgttt attagaataa cggatattta aaagggcgtg 2940

aaaaggttta tccgttcgtc catttgtatg tgggt 2975

<210> 17

<211> 2682

<212> DNA

<213> 人工序列(Artificial sequence)

<400> 17

gatcacaggc agcaacgctc tgtcatcgtt acaatcaaca tgctaccctc cgcgagatca 60

tccgtgtttc aaacccggca gcttagttgc cgttcttccg aatagcatcg gtaacatgag 120

caaagtctgc cgccttacaa cggctctccc gctgacgccg tcccggactg atgggctgcc 180

tgtatcgagt ggtgattttg tgccgagctg ccggtcgggg agctgttggc tggctggtgg 240

caggatatat tgtggtgtaa acaaattgac gcttagacaa cttaataaca cattgcggac 300

gtttttaatg tactgaatta acgccgaatt aattcggggg atctggattt tagtactgga 360

ttttggtttt aggaattaga aattttattg atagaagtat tttacaaata caaatacata 420

ctaagggttt cttatatgct caacacatga gcgaaaccct ataggaaccc taattccctt 480

atctgggaac tactcacaca ttattatgga gaaactcgac ggtatcgata agcttgatcc 540

atgcctcaca tgttaatgta ctaccaatgg aggcttccat gcctcacatg ttcatgtaca 600

catttatgat taggaaactt tttaatatat tttatagatt tcttatccat catataaaaa 660

tacataatta atcatacgat tttgagatac atattctgac gtatcaaatt ctaattaaat 720

tttaaaatat tttagtgacg tatcaaattc taattaaatt ttaaaatatt ttagagacgt 780

attttcgtaa caatttaaaa tgtatattat agatcacatt cataggtcat tttataattt 840

aaaatattat ggagatgcat cttcgtttat ttttacggag atatattttc gtaatttatc 900

ataatagaat tgttcatgct atattttgtt tatgtttgct cagatgaaga tttaaacctt 960

acaagcaatg tgcaaaaaat gacgtacata aatttagatg gtccaaaaat gttataaata 1020

aaagatcaag aagtgtcaaa aaaagtcaaa aacaacgata gagtagtata atgtcaaaat 1080

aaaataaaat ccatgacact actactatta tatattaatg cactaatgtg tatgtctaac 1140

tacatcgcct ctgcctcctc tgtcagttat gtctcgtagg ccatcaatcc cccgtcctcc 1200

gacgttgtct ccggtacatc aatgtcccat gtgcctacgt catgatggca tttaggacat 1260

gtctcacatc agccagatca gcaagataca tttgtcaatg tctatctacg caatctccac 1320

aatgcgacga catataggca agacatcctc aacataattt agttgtgcat gcttctcctc 1380

tagtatctcc cgatgagttg atcgaattaa ttcctgcagc ccttggcaag ctgctctagc 1440

caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca 1500

ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc 1560

attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga 1620

gcggataaca atttcacaca ggaaacagct atgacatgat tacgaattcg agctcggtac 1680

ccggggatcc tctagagtcg acctgcaggc atgcaagctt ggcactggcc gtcgttttac 1740

aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc 1800

ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc 1860

gcagcctgaa tggcgaatgc tagagcagct tgagcttgga tcagattgtc gtttcccgcc 1920

ttcagtttaa ttcgatccat gcctcacatg ttaatgtact accaatggag ggctgtacac 1980

atttatgatt acgaaatttt ttaatatatt ttatagattt cttatgcatc atacaaaaat 2040

acataattat tcgtaacatt ttggagatac atattcagat gcatcaaatt ctaattaaac 2100

gttaaaatat tttggagacg tatcttcgta acaatttaaa acctatacta tacatcacat 2160

tcgaaggtca ttttataatt taaaatatta tggagatgca tcttcgttta tgtttgctca 2220

gatgaagatt taaaccttac aaacaatatg taaaaaatga cgtacataaa ttcagatagt 2280

ccaaaagtgt catatataaa taaagatcaa taagtgtcaa aaaaagtcaa gaacaacgat 2340

agagtagcat aatgtcaaaa taaaataaaa tccatgacac tactactatt atatattaat 2400

gcactaatgt gtatgtctaa ctacatcgtc tctgcctcct ctgtcagtta tgtctcgtaa 2460

gccatcaatc ccccgtcctc cggcgttgtc tccggtatat caatgtcccc atgtgcctac 2520

gtcatgatgg catctaggac atgtctcaca tcagacacat taggaaagat acatttgcca 2580

atgtatatct gcgcaatctc cacaatgcaa cgacatatag gcaagacatc ctcaacataa 2640

tttagttgtg catgcttctc ctctagtatc tcccgatgag tt 2682

<210> 18

<211> 8749

<212> DNA

<213> 人工序列(Artificial sequence)

<400> 18

catgccaacc acagggttcc cctcgggatc aaagtacttt gatccaaccc ctccgctgct 60

atagtgcagt cggcttctga cgttcagtgc agccgtcttc tgaaaacgac atgtcgcaca 120

agtcctaagt tacgcgacag gctgccgccc tgcccttttc ctggcgtttt cttgtcgcgt 180

gttttagtcg cataaagtag aatacttgcg actagaaccg gagacattac gccatgaaca 240

agagcgccgc cgctggcctg ctgggctatg cccgcgtcag caccgacgac caggacttga 300

ccaaccaacg ggccgaactg cacgcggccg gctgcaccaa gctgttttcc gagaagatca 360

ccggcaccag gcgcgaccgc ccggagctgg ccaggatgct tgaccaccta cgccctggcg 420

acgttgtgac agtgaccagg ctagaccgcc tggcccgcag cacccgcgac ctactggaca 480

ttgccgagcg catccaggag gccggcgcgg gcctgcgtag cctggcagag ccgtgggccg 540

acaccaccac gccggccggc cgcatggtgt tgaccgtgtt cgccggcatt gccgagttcg 600

agcgttccct aatcatcgac cgcacccgga gcgggcgcga ggccgccaag gcccgaggcg 660

tgaagtttgg cccccgccct accctcaccc cggcacagat cgcgcacgcc cgcgagctga 720

tcgaccagga aggccgcacc gtgaaagagg cggctgcact gcttggcgtg catcgctcga 780

ccctgtaccg cgcacttgag cgcagcgagg aagtgacgcc caccgaggcc aggcggcgcg 840

gtgccttccg tgaggacgca ttgaccgagg ccgacgccct ggcggccgcc gagaatgaac 900

gccaagagga acaagcatga aaccgcacca ggacggccag gacgaaccgt ttttcattac 960

cgaagagatc gaggcggaga tgatcgcggc cgggtacgtg ttcgagccgc ccgcgcacgt 1020

ctcaaccgtg cggctgcatg aaatcctggc cggtttgtct gatgccaagc tggcggcctg 1080

gccggccagc ttggccgctg aagaaaccga gcgccgccgt ctaaaaaggt gatgtgtatt 1140

tgagtaaaac agcttgcgtc atgcggtcgc tgcgtatatg atgcgatgag taaataaaca 1200

aatacgcaag gggaacgcat gaaggttatc gctgtactta accagaaagg cgggtcaggc 1260

aagacgacca tcgcaaccca tctagcccgc gccctgcaac tcgccggggc cgatgttctg 1320

ttagtcgatt ccgatcccca gggcagtgcc cgcgattggg cggccgtgcg ggaagatcaa 1380

ccgctaaccg ttgtcggcat cgaccgcccg acgattgacc gcgacgtgaa ggccatcggc 1440

cggcgcgact tcgtagtgat cgacggagcg ccccaggcgg cggacttggc tgtgtccgcg 1500

atcaaggcag ccgacttcgt gctgattccg gtgcagccaa gcccttacga catatgggcc 1560

accgccgacc tggtggagct ggttaagcag cgcattgagg tcacggatgg aaggctacaa 1620

gcggcctttg tcgtgtcgcg ggcgatcaaa ggcacgcgca tcggcggtga ggttgccgag 1680

gcgctggccg ggtacgagct gcccattctt gagtcccgta tcacgcagcg cgtgagctac 1740

ccaggcactg ccgccgccgg cacaaccgtt cttgaatcag aacccgaggg cgacgctgcc 1800

cgcgaggtcc aggcgctggc cgctgaaatt aaatcaaaac tcatttgagt taatgaggta 1860

aagagaaaat gagcaaaagc acaaacacgc taagtgccgg ccgtccgagc gcacgcagca 1920

gcaaggctgc aacgttggcc agcctggcag acacgccagc catgaagcgg gtcaactttc 1980

agttgccggc ggaggatcac accaagctga agatgtacgc ggtacgccaa ggcaagacca 2040

ttaccgagct gctatctgaa tacatcgcgc agctaccaga gtaaatgagc aaatgaataa 2100

atgagtagat gaattttagc ggctaaagga ggcggcatgg aaaatcaaga acaaccaggc 2160

accgacgccg tggaatgccc catgtgtgga ggaacgggcg gttggccagg cgtaagcggc 2220

tgggttgtct gccggccctg caatggcact ggaaccccca agcccgagga atcggcgtga 2280

cggtcgcaaa ccatccggcc cggtacaaat cggcgcggcg ctgggtgatg acctggtgga 2340

gaagttgaag gccgcgcagg ccgcccagcg gcaacgcatc gaggcagaag cacgccccgg 2400

tgaatcgtgg caagcggccg ctgatcgaat ccgcaaagaa tcccggcaac cgccggcagc 2460

cggtgcgccg tcgattagga agccgcccaa gggcgacgag caaccagatt ttttcgttcc 2520

gatgctctat gacgtgggca cccgcgatag tcgcagcatc atggacgtgg ccgttttccg 2580

tctgtcgaag cgtgaccgac gagctggcga ggtgatccgc tacgagcttc cagacgggca 2640

cgtagaggtt tccgcagggc cggccggcat ggccagtgtg tgggattacg acctggtact 2700

gatggcggtt tcccatctaa ccgaatccat gaaccgatac cgggaaggga agggagacaa 2760

gcccggccgc gtgttccgtc cacacgttgc ggacgtactc aagttctgcc ggcgagccga 2820

tggcggaaag cagaaagacg acctggtaga aacctgcatt cggttaaaca ccacgcacgt 2880

tgccatgcag cgtacgaaga aggccaagaa cggccgcctg gtgacggtat ccgagggtga 2940

agccttgatt agccgctaca agatcgtaaa gagcgaaacc gggcggccgg agtacatcga 3000

gatcgagcta gctgattgga tgtaccgcga gatcacagaa ggcaagaacc cggacgtgct 3060

gacggttcac cccgattact ttttgatcga tcccggcatc ggccgttttc tctaccgcct 3120

ggcacgccgc gccgcaggca aggcagaagc cagatggttg ttcaagacga tctacgaacg 3180

cagtggcagc gccggagagt tcaagaagtt ctgtttcacc gtgcgcaagc tgatcgggtc 3240

aaatgacctg ccggagtacg atttgaagga ggaggcgggg caggctggcc cgatcctagt 3300

catgcgctac cgcaacctga tcgagggcga agcatccgcc ggttcctaat gtacggagca 3360

gatgctaggg caaattgccc tagcagggga aaaaggtcga aaaggtctct ttcctgtgga 3420

tagcacgtac attgggaacc caaagccgta cattgggaac cggaacccgt acattgggaa 3480

cccaaagccg tacattggga accggtcaca catgtaagtg actgatataa aagagaaaaa 3540

aggcgatttt tccgcctaaa actctttaaa acttattaaa actcttaaaa cccgcctggc 3600

ctgtgcataa ctgtctggcc agcgcacagc cgaagagctg caaaaagcgc ctacccttcg 3660

gtcgctgcgc tccctacgcc ccgccgcttc gcgtcggcct atcgcggccg ctggccgctc 3720

aaaaatggct ggcctacggc caggcaatct accagggcgc ggacaagccg cgccgtcgcc 3780

actcgaccgc cggcgcccac atcaaggcac cctgcctcgc gcgtttcggt gatgacggtg 3840

aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg 3900

ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca 3960

tgacccagtc acgtagcgat agcggagtgt atactggctt aactatgcgg catcagagca 4020

gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa 4080

ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg 4140

gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg 4200

ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa 4260

ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 4320

acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 4380

tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 4440

ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc 4500

ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg 4560

ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc 4620

actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga 4680

gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc 4740

tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac 4800

caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 4860

atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 4920

acgttaaggg attttggtca tgcattctag gtactaaaac aattcatcca gtaaaatata 4980

atattttatt ttctcccaat caggcttgat ccccagtaag tcaaaaaata gctcgacata 5040

ctgttcttcc ccgatatcct ccctgatcga ccggacgcag aaggcaatgt cataccactt 5100

gtccgccctg ccgcttctcc caagatcaat aaagccactt actttgccat ctttcacaaa 5160

gatgttgctg tctcccaggt cgccgtggga aaagacaagt tcctcttcgg gcttttccgt 5220

ctttaaaaaa tcatacagct cgcgcggatc tttaaatgga gtgtcttctt cccagttttc 5280

gcaatccaca tcggccagat cgttattcag taagtaatcc aattcggcta agcggctgtc 5340

taagctattc gtatagggac aatccgatat gtcgatggag tgaaagagcc tgatgcactc 5400

cgcatacagc tcgataatct tttcagggct ttgttcatct tcatactctt ccgagcaaag 5460

gacgccatcg gcctcactca tgagcagatt gctccagcca tcatgccgtt caaagtgcag 5520

gacctttgga acaggcagct ttccttccag ccatagcatc atgtcctttt cccgttccac 5580

atcataggtg gtccctttat accggctgtc cgtcattttt aaatataggt tttcattttc 5640

tcccaccagc ttatatacct tagcaggaga cattccttcc gtatctttta cgcagcggta 5700

tttttcgatc agttttttca attccggtga tattctcatt ttagccattt attatttcct 5760

tcctcttttc tacagtattt aaagataccc caagaagcta attataacaa gacgaactcc 5820

aattcactgt tccttgcatt ctaaaacctt aaataccaga aaacagcttt ttcaaagttg 5880

ttttcaaagt tggcgtataa catagtatcg acggagccga ttttgaaacc ggtgatcaca 5940

ggcagcaacg ctctgtcatc gttacaatca acatgctacc ctccgcgaga tcatccgtgt 6000

ttcaaacccg gcagcttagt tgccgttctt ccgaatagca tcggtaacat gagcaaagtc 6060

tgccgcctta caacggctct cccgctgacg ccgtcccgga ctgatgggct gcctgtatcg 6120

agtggtgatt ttgtgccgag ctgccggtcg gggagctgtt ggctggctgg tggcaggata 6180

tattgtggtg taaacaaatt gacgcttaga caacttaata acacattgcg gacgttttta 6240

atgtactgaa ttaacgccga attaattcgg gggatctgga ttttagtact ggattttggt 6300

tttaggaatt agaaatttta ttgatagaag tattttacaa atacaaatac atactaaggg 6360

tttcttatat gctcaacaca tgagcgaaac cctataggaa ccctaattcc cttatctggg 6420

aactactcac acattattat ggagaaactc gacggtatcg ataagcttga tccatgcctc 6480

acatgttaat gtactaccaa tggaggcttc catgcctcac atgttcatgt acacatttat 6540

gattaggaaa ctttttaata tattttatag atttcttatc catcatataa aaatacataa 6600

ttaatcatac gattttgaga tacatattct gacgtatcaa attctaatta aattttaaaa 6660

tattttagtg acgtatcaaa ttctaattaa attttaaaat attttagaga cgtattttcg 6720

taacaattta aaatgtatat tatagatcac attcataggt cattttataa tttaaaatat 6780

tatggagatg catcttcgtt tatttttacg gagatatatt ttcgtaattt atcataatag 6840

aattgttcat gctatatttt gtttatgttt gctcagatga agatttaaac cttacaagca 6900

atgtgcaaaa aatgacgtac ataaatttag atggtccaaa aatgttataa ataaaagatc 6960

aagaagtgtc aaaaaaagtc aaaaacaacg atagagtagt ataatgtcaa aataaaataa 7020

aatccatgac actactacta ttatatatta atgcactaat gtgtatgtct aactacatcg 7080

cctctgcctc ctctgtcagt tatgtctcgt aggccatcaa tcccccgtcc tccgacgttg 7140

tctccggtac atcaatgtcc catgtgccta cgtcatgatg gcatttagga catgtctcac 7200

atcagccaga tcagcaagat acatttgtca atgtctatct acgcaatctc cacaatgcga 7260

cgacatatag gcaagacatc ctcaacataa tttagttgtg catgcttctc ctctagtatc 7320

tcccgatgag ttgatcgaat taattcctgc agcccttggc aagctgctct agccaatacg 7380

caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc 7440

cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc 7500

accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata 7560

acaatttcac acaggaaaca gctatgacat gattacgaat tcgagctcgg tacccgggga 7620

tcctctagag tcgacctgca ggcatgcaag cttggcactg gccgtcgttt tacaacgtcg 7680

tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc 7740

cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct 7800

gaatggcgaa tgctagagca gcttgagctt ggatcagatt gtcgtttccc gccttcagtt 7860

taattcgatc catgcctcac atgttaatgt actaccaatg gagggctgta cacatttatg 7920

attacgaaat tttttaatat attttataga tttcttatgc atcatacaaa aatacataat 7980

tattcgtaac attttggaga tacatattca gatgcatcaa attctaatta aacgttaaaa 8040

tattttggag acgtatcttc gtaacaattt aaaacctata ctatacatca cattcgaagg 8100

tcattttata atttaaaata ttatggagat gcatcttcgt ttatgtttgc tcagatgaag 8160

atttaaacct tacaaacaat atgtaaaaaa tgacgtacat aaattcagat agtccaaaag 8220

tgtcatatat aaataaagat caataagtgt caaaaaaagt caagaacaac gatagagtag 8280

cataatgtca aaataaaata aaatccatga cactactact attatatatt aatgcactaa 8340

tgtgtatgtc taactacatc gtctctgcct cctctgtcag ttatgtctcg taagccatca 8400

atcccccgtc ctccggcgtt gtctccggta tatcaatgtc cccatgtgcc tacgtcatga 8460

tggcatctag gacatgtctc acatcagaca cattaggaaa gatacatttg ccaatgtata 8520

tctgcgcaat ctccacaatg caacgacata taggcaagac atcctcaaca taatttagtt 8580

gtgcatgctt ctcctctagt atctcccgat gagttgatca agcttatcga aactatcagt 8640

gtttgacagg atatattggc gggtaaacct aagagaaaag agcgtttatt agaataacgg 8700

atatttaaaa gggcgtgaaa aggtttatcc gttcgtccat ttgtatgtg 8749

<210> 19

<211> 3783

<212> DNA

<213> 人工序列(Artificial sequence)

<400> 19

cgggcccccc ctcgaggtcg acggtatcga taagcttgat atcgaattcc tgcagctcac 60

tcttttaaaa gctcagttca gttcctgacg aaagtgctta gaacgtcgaa gtagttgggg 120

aaggtcttgc gggtgcaacc agggtccctg atcgtcacgg gcacgtcggc gcaggcagcg 180

agggagaagg ccatggccat cctgtgatca tcgtaggtgt cgattgccgt gatgttcagc 240

ttctccggtg gggtgatgat gcagtagtca ggaccttctt caaccgatgc tcccagcttt 300

gttagctcgg tccgaattgc aaccatcctt tcggtttcct ttactctcca ggaagccaca 360

tctctgatag cagttggacc atcagcgaag agtgcaacaa cggcaagggt catggcaaca 420

tcaggcattt tgttcatgtt gacatcaaca gctttcaggt gtttcttccc ataaggctca 480

cgtggtggac cagttacggt tacactggtg tcagtccatg taacctttgc tcccatcatc 540

tcaagtacct cagcaaattt gacatcaccc tgcaaactgg tcgtaccaca accttgaact 600

gtcacagtgc ctccagtgat tgcagcacca gccaagaaat agctcgcgct tgaggcatca 660

ccttcaacat aggcatttcc aggagatttg tacttctgcc ctcccttaat atagaatctg 720

tcccaactat cagaatgctc tgccttcaca ccaaaacgct ccatcaatct caatgtcatt 780

tcaacgtaag gaatggagat tagtttgtca atgatttcga tctccacatc cccaagggcc 840

aaaggagcag ccatcagcaa ggcactcaag tactgactgc tgatggaacc agagagctta 900

accttgccac caggaagtcc tccaattccc ttgacacgaa caggtgggca ttcagtgcca 960

aggaaacagt cgacatccgc accaagttgt ttcaacccga caaccaagtc accaatcggt 1020

ctctccctca ttcgtggcac tccatcaagc acataagttg catttccacc agcagcagtc 1080

acggctgctg tcaatagtcg cattgcagtt ccagcgttcc ccaagaagag ttgcacttcc 1140

tctttcgcat ccttctcaac aggaaacttg ccaccacagc caacgactac agctcttttt 1200

gcaactttat ctgcttccac agagagcccg agggctttca gggcctcaag catgtagtga 1260

acatcctcac tgttcagcaa gttgtccacc actgttgtgc cctcggagag ggcggagagg 1320

aggaggatcc tgttggagag cgacttggac cctggcagct gaaccgcccc ggagatctcc 1380

ctgatgggct ggagcacgat ctcctccgcc ttcgccgccg gcgctgccac cgacgacgac 1440

gacgcggacg ccaccaccac cgcctcccgc cgcccccgcg cccgcacccg cacccgcatc 1500

cccccgcgcg ccgcggcggg cagccgcagc tgcttccgcg acgagaacgc cgccgacgcc 1560

gccacggcct ggtccaggga caccgccgcc gcagccgcgg cgttggacgc catggtcgcc 1620

gccattctag agcggccgct ctagaactag tggatccgaa gtaacaccaa acaacagggt 1680

gagcatcgac aaaagaaaca gtaccaagca aataaatagc gtatgaaggc agggctaaaa 1740

aaatccacat atagctgctg catatgccat catccaagta tatcaagatc aaaataatta 1800

taaaacatac ttgtttatta taatagatag gtactcaagg ttagagcata tgaatagatg 1860

ctgcatatgc catcatgtat atgcatcagt aaaacccaca tcaacatgta tacctatcct 1920

agatcgatat ttccatccat cttaaactcg taactatgaa gatgtatgac acacacatac 1980

agttccaaaa ttaataaata caccaggtag tttgaaacag tattctactc cgatctagaa 2040

cgaatgaacg accgcccaac cacaccacat catcacaacc aagcgaacaa aaagcatctc 2100

tgtatatgca tcagtaaaac ccgcatcaac atgtatacct atcctagatc gatatttcca 2160

tccatcatct tcaattcgta actatgaata tgtatggcac acacatacag atccaaaatt 2220

aataaatcca ccaggtagtt tgaaacagaa ttctactccg atctagaacg accgcccaac 2280

cagaccacat catcacaacc aagacaaaaa aaagcatgaa aagatgaccc gacaaacaag 2340

tgcacggcat atattgaaat aaaggaaaag ggcaaaccaa accctatgca acgaaacaaa 2400

aaaaatcatg aaatcgatcc cgtctgcgga acggctagag ccatcccagg attccccaaa 2460

gagaaacact ggcaagttag caatcagaac gtgtctgacg tacaggtcgc atccgtgtac 2520

gaacgctagc agcacggatc taacacaaac acggatctaa cacaaacatg aacagaagta 2580

gaactaccgg gccctaacca tggaccggaa cgccgatcta gagaaggtag agaggggggg 2640

ggggggagga cgagcggcgt accttgaagc ggaggtgccg acgggtggat ttgggggaga 2700

tctggttgtg tgtgtgtgcg ctccgaacaa cacgaggttg gggaaagagg gtgtggaggg 2760

ggtgtctatt tattacggcg ggcgaggaag ggaaagcgaa ggagcggtgg gaaaggaatc 2820

ccccgtagct gccgtgccgt gagaggagga ggaggccgcc tgccgtgccg gctcacgtct 2880

gccgctccgc cacgcaattt ctggatgccg acagcggagc aagtccaacg gtggagcgga 2940

actctcgaga ggggtccaga ggcagcgaca gagatgccgt gccgtctgct tcgcttggcc 3000

cgacgcgacg ctgctggttc gctggttggt gtccgttaga ctcgtcgacg gcgtttaaca 3060

ggctggcatt atctactcga aacaagaaaa atgtttcctt agttttttta atttcttaaa 3120

gggtatttgt ttaattttta gtcactttat tttattctat tttatatcta aattattaaa 3180

taaaaaaact aaaatagagt tttagttttc ttaatttaga ggctaaaata gaataaaata 3240

gatgtactaa aaaaattagt ctataaaaac cattaaccct aaaccctaaa tggatgtact 3300

aataaaatgg atgaagtatt atataggtga agctatttgc aaaaaaaaag gagaacacat 3360

gcacactaaa aagataaaac tgtagagtcc tgttgtcaaa atactcaatt gtcctttaga 3420

ccatgtctaa ctgttcattt atatgattct ctaaaacact gatattattg tagtactata 3480

gattatatta ttcgtagagt aaagtttaaa tatatgtata aagatagata aactgcactt 3540

caacaagctt ggcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta 3600

cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg 3660

cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgc tagagcagct 3720

tgagcttgga tcagattgtc gtttcccgcc ttcagtttgg ggatcctcta gagtcgacct 3780

gca 3783

<210> 20

<211> 12505

<212> DNA

<213> 人工序列(Artificial sequence)

<400> 20

catgccaacc acagggttcc cctcgggatc aaagtacttt gatccaaccc ctccgctgct 60

atagtgcagt cggcttctga cgttcagtgc agccgtcttc tgaaaacgac atgtcgcaca 120

agtcctaagt tacgcgacag gctgccgccc tgcccttttc ctggcgtttt cttgtcgcgt 180

gttttagtcg cataaagtag aatacttgcg actagaaccg gagacattac gccatgaaca 240

agagcgccgc cgctggcctg ctgggctatg cccgcgtcag caccgacgac caggacttga 300

ccaaccaacg ggccgaactg cacgcggccg gctgcaccaa gctgttttcc gagaagatca 360

ccggcaccag gcgcgaccgc ccggagctgg ccaggatgct tgaccaccta cgccctggcg 420

acgttgtgac agtgaccagg ctagaccgcc tggcccgcag cacccgcgac ctactggaca 480

ttgccgagcg catccaggag gccggcgcgg gcctgcgtag cctggcagag ccgtgggccg 540

acaccaccac gccggccggc cgcatggtgt tgaccgtgtt cgccggcatt gccgagttcg 600

agcgttccct aatcatcgac cgcacccgga gcgggcgcga ggccgccaag gcccgaggcg 660

tgaagtttgg cccccgccct accctcaccc cggcacagat cgcgcacgcc cgcgagctga 720

tcgaccagga aggccgcacc gtgaaagagg cggctgcact gcttggcgtg catcgctcga 780

ccctgtaccg cgcacttgag cgcagcgagg aagtgacgcc caccgaggcc aggcggcgcg 840

gtgccttccg tgaggacgca ttgaccgagg ccgacgccct ggcggccgcc gagaatgaac 900

gccaagagga acaagcatga aaccgcacca ggacggccag gacgaaccgt ttttcattac 960

cgaagagatc gaggcggaga tgatcgcggc cgggtacgtg ttcgagccgc ccgcgcacgt 1020

ctcaaccgtg cggctgcatg aaatcctggc cggtttgtct gatgccaagc tggcggcctg 1080

gccggccagc ttggccgctg aagaaaccga gcgccgccgt ctaaaaaggt gatgtgtatt 1140

tgagtaaaac agcttgcgtc atgcggtcgc tgcgtatatg atgcgatgag taaataaaca 1200

aatacgcaag gggaacgcat gaaggttatc gctgtactta accagaaagg cgggtcaggc 1260

aagacgacca tcgcaaccca tctagcccgc gccctgcaac tcgccggggc cgatgttctg 1320

ttagtcgatt ccgatcccca gggcagtgcc cgcgattggg cggccgtgcg ggaagatcaa 1380

ccgctaaccg ttgtcggcat cgaccgcccg acgattgacc gcgacgtgaa ggccatcggc 1440

cggcgcgact tcgtagtgat cgacggagcg ccccaggcgg cggacttggc tgtgtccgcg 1500

atcaaggcag ccgacttcgt gctgattccg gtgcagccaa gcccttacga catatgggcc 1560

accgccgacc tggtggagct ggttaagcag cgcattgagg tcacggatgg aaggctacaa 1620

gcggcctttg tcgtgtcgcg ggcgatcaaa ggcacgcgca tcggcggtga ggttgccgag 1680

gcgctggccg ggtacgagct gcccattctt gagtcccgta tcacgcagcg cgtgagctac 1740

ccaggcactg ccgccgccgg cacaaccgtt cttgaatcag aacccgaggg cgacgctgcc 1800

cgcgaggtcc aggcgctggc cgctgaaatt aaatcaaaac tcatttgagt taatgaggta 1860

aagagaaaat gagcaaaagc acaaacacgc taagtgccgg ccgtccgagc gcacgcagca 1920

gcaaggctgc aacgttggcc agcctggcag acacgccagc catgaagcgg gtcaactttc 1980

agttgccggc ggaggatcac accaagctga agatgtacgc ggtacgccaa ggcaagacca 2040

ttaccgagct gctatctgaa tacatcgcgc agctaccaga gtaaatgagc aaatgaataa 2100

atgagtagat gaattttagc ggctaaagga ggcggcatgg aaaatcaaga acaaccaggc 2160

accgacgccg tggaatgccc catgtgtgga ggaacgggcg gttggccagg cgtaagcggc 2220

tgggttgtct gccggccctg caatggcact ggaaccccca agcccgagga atcggcgtga 2280

cggtcgcaaa ccatccggcc cggtacaaat cggcgcggcg ctgggtgatg acctggtgga 2340

gaagttgaag gccgcgcagg ccgcccagcg gcaacgcatc gaggcagaag cacgccccgg 2400

tgaatcgtgg caagcggccg ctgatcgaat ccgcaaagaa tcccggcaac cgccggcagc 2460

cggtgcgccg tcgattagga agccgcccaa gggcgacgag caaccagatt ttttcgttcc 2520

gatgctctat gacgtgggca cccgcgatag tcgcagcatc atggacgtgg ccgttttccg 2580

tctgtcgaag cgtgaccgac gagctggcga ggtgatccgc tacgagcttc cagacgggca 2640

cgtagaggtt tccgcagggc cggccggcat ggccagtgtg tgggattacg acctggtact 2700

gatggcggtt tcccatctaa ccgaatccat gaaccgatac cgggaaggga agggagacaa 2760

gcccggccgc gtgttccgtc cacacgttgc ggacgtactc aagttctgcc ggcgagccga 2820

tggcggaaag cagaaagacg acctggtaga aacctgcatt cggttaaaca ccacgcacgt 2880

tgccatgcag cgtacgaaga aggccaagaa cggccgcctg gtgacggtat ccgagggtga 2940

agccttgatt agccgctaca agatcgtaaa gagcgaaacc gggcggccgg agtacatcga 3000

gatcgagcta gctgattgga tgtaccgcga gatcacagaa ggcaagaacc cggacgtgct 3060

gacggttcac cccgattact ttttgatcga tcccggcatc ggccgttttc tctaccgcct 3120

ggcacgccgc gccgcaggca aggcagaagc cagatggttg ttcaagacga tctacgaacg 3180

cagtggcagc gccggagagt tcaagaagtt ctgtttcacc gtgcgcaagc tgatcgggtc 3240

aaatgacctg ccggagtacg atttgaagga ggaggcgggg caggctggcc cgatcctagt 3300

catgcgctac cgcaacctga tcgagggcga agcatccgcc ggttcctaat gtacggagca 3360

gatgctaggg caaattgccc tagcagggga aaaaggtcga aaaggtctct ttcctgtgga 3420

tagcacgtac attgggaacc caaagccgta cattgggaac cggaacccgt acattgggaa 3480

cccaaagccg tacattggga accggtcaca catgtaagtg actgatataa aagagaaaaa 3540

aggcgatttt tccgcctaaa actctttaaa acttattaaa actcttaaaa cccgcctggc 3600

ctgtgcataa ctgtctggcc agcgcacagc cgaagagctg caaaaagcgc ctacccttcg 3660

gtcgctgcgc tccctacgcc ccgccgcttc gcgtcggcct atcgcggccg ctggccgctc 3720

aaaaatggct ggcctacggc caggcaatct accagggcgc ggacaagccg cgccgtcgcc 3780

actcgaccgc cggcgcccac atcaaggcac cctgcctcgc gcgtttcggt gatgacggtg 3840

aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg 3900

ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca 3960

tgacccagtc acgtagcgat agcggagtgt atactggctt aactatgcgg catcagagca 4020

gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa 4080

ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg 4140

gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg 4200

ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa 4260

ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 4320

acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 4380

tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 4440

ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc 4500

ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg 4560

ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc 4620

actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga 4680

gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc 4740

tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac 4800

caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 4860

atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 4920

acgttaaggg attttggtca tgcattctag gtactaaaac aattcatcca gtaaaatata 4980

atattttatt ttctcccaat caggcttgat ccccagtaag tcaaaaaata gctcgacata 5040

ctgttcttcc ccgatatcct ccctgatcga ccggacgcag aaggcaatgt cataccactt 5100

gtccgccctg ccgcttctcc caagatcaat aaagccactt actttgccat ctttcacaaa 5160

gatgttgctg tctcccaggt cgccgtggga aaagacaagt tcctcttcgg gcttttccgt 5220

ctttaaaaaa tcatacagct cgcgcggatc tttaaatgga gtgtcttctt cccagttttc 5280

gcaatccaca tcggccagat cgttattcag taagtaatcc aattcggcta agcggctgtc 5340

taagctattc gtatagggac aatccgatat gtcgatggag tgaaagagcc tgatgcactc 5400

cgcatacagc tcgataatct tttcagggct ttgttcatct tcatactctt ccgagcaaag 5460

gacgccatcg gcctcactca tgagcagatt gctccagcca tcatgccgtt caaagtgcag 5520

gacctttgga acaggcagct ttccttccag ccatagcatc atgtcctttt cccgttccac 5580

atcataggtg gtccctttat accggctgtc cgtcattttt aaatataggt tttcattttc 5640

tcccaccagc ttatatacct tagcaggaga cattccttcc gtatctttta cgcagcggta 5700

tttttcgatc agttttttca attccggtga tattctcatt ttagccattt attatttcct 5760

tcctcttttc tacagtattt aaagataccc caagaagcta attataacaa gacgaactcc 5820

aattcactgt tccttgcatt ctaaaacctt aaataccaga aaacagcttt ttcaaagttg 5880

ttttcaaagt tggcgtataa catagtatcg acggagccga ttttgaaacc ggtgatcaca 5940

ggcagcaacg ctctgtcatc gttacaatca acatgctacc ctccgcgaga tcatccgtgt 6000

ttcaaacccg gcagcttagt tgccgttctt ccgaatagca tcggtaacat gagcaaagtc 6060

tgccgcctta caacggctct cccgctgacg ccgtcccgga ctgatgggct gcctgtatcg 6120

agtggtgatt ttgtgccgag ctgccggtcg gggagctgtt ggctggctgg tggcaggata 6180

tattgtggtg taaacaaatt gacgcttaga caacttaata acacattgcg gacgttttta 6240

atgtactgaa ttaacgccga attaattcgg gggatctgga ttttagtact ggattttggt 6300

tttaggaatt agaaatttta ttgatagaag tattttacaa atacaaatac atactaaggg 6360

tttcttatat gctcaacaca tgagcgaaac cctataggaa ccctaattcc cttatctggg 6420

aactactcac acattattat ggagaaactc gacggtatcg ataagcttga tccatgcctc 6480

acatgttaat gtactaccaa tggaggcttc catgcctcac atgttcatgt acacatttat 6540

gattaggaaa ctttttaata tattttatag atttcttatc catcatataa aaatacataa 6600

ttaatcatac gattttgaga tacatattct gacgtatcaa attctaatta aattttaaaa 6660

tattttagtg acgtatcaaa ttctaattaa attttaaaat attttagaga cgtattttcg 6720

taacaattta aaatgtatat tatagatcac attcataggt cattttataa tttaaaatat 6780

tatggagatg catcttcgtt tatttttacg gagatatatt ttcgtaattt atcataatag 6840

aattgttcat gctatatttt gtttatgttt gctcagatga agatttaaac cttacaagca 6900

atgtgcaaaa aatgacgtac ataaatttag atggtccaaa aatgttataa ataaaagatc 6960

aagaagtgtc aaaaaaagtc aaaaacaacg atagagtagt ataatgtcaa aataaaataa 7020

aatccatgac actactacta ttatatatta atgcactaat gtgtatgtct aactacatcg 7080

cctctgcctc ctctgtcagt tatgtctcgt aggccatcaa tcccccgtcc tccgacgttg 7140

tctccggtac atcaatgtcc catgtgccta cgtcatgatg gcatttagga catgtctcac 7200

atcagccaga tcagcaagat acatttgtca atgtctatct acgcaatctc cacaatgcga 7260

cgacatatag gcaagacatc ctcaacataa tttagttgtg catgcttctc ctctagtatc 7320

tcccgatgag ttgatcgaat taattcctgc agcccttggc aagctgctct agccaatacg 7380

caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc 7440

cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc 7500

accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata 7560

acaatttcac acaggaaaca gctatgacat gattacgaat tcgagctcgg taccgggccc 7620

cccctcgagg tcgacggtat cgataagctt gatatcgaat tcctgcagct cactctttta 7680

aaagctcagt tcagttcctg acgaaagtgc ttagaacgtc gaagtagttg gggaaggtct 7740

tgcgggtgca accagggtcc ctgatcgtca cgggcacgtc ggcgcaggca gcgagggaga 7800

aggccatggc catcctgtga tcatcgtagg tgtcgattgc cgtgatgttc agcttctccg 7860

gtggggtgat gatgcagtag tcaggacctt cttcaaccga tgctcccagc tttgttagct 7920

cggtccgaat tgcaaccatc ctttcggttt cctttactct ccaggaagcc acatctctga 7980

tagcagttgg accatcagcg aagagtgcaa caacggcaag ggtcatggca acatcaggca 8040

ttttgttcat gttgacatca acagctttca ggtgtttctt cccataaggc tcacgtggtg 8100

gaccagttac ggttacactg gtgtcagtcc atgtaacctt tgctcccatc atctcaagta 8160

cctcagcaaa tttgacatca ccctgcaaac tggtcgtacc acaaccttga actgtcacag 8220

tgcctccagt gattgcagca ccagccaaga aatagctcgc gcttgaggca tcaccttcaa 8280

cataggcatt tccaggagat ttgtacttct gccctccctt aatatagaat ctgtcccaac 8340

tatcagaatg ctctgccttc acaccaaaac gctccatcaa tctcaatgtc atttcaacgt 8400

aaggaatgga gattagtttg tcaatgattt cgatctccac atccccaagg gccaaaggag 8460

cagccatcag caaggcactc aagtactgac tgctgatgga accagagagc ttaaccttgc 8520

caccaggaag tcctccaatt cccttgacac gaacaggtgg gcattcagtg ccaaggaaac 8580

agtcgacatc cgcaccaagt tgtttcaacc cgacaaccaa gtcaccaatc ggtctctccc 8640

tcattcgtgg cactccatca agcacataag ttgcatttcc accagcagca gtcacggctg 8700

ctgtcaatag tcgcattgca gttccagcgt tccccaagaa gagttgcact tcctctttcg 8760

catccttctc aacaggaaac ttgccaccac agccaacgac tacagctctt tttgcaactt 8820

tatctgcttc cacagagagc ccgagggctt tcagggcctc aagcatgtag tgaacatcct 8880

cactgttcag caagttgtcc accactgttg tgccctcgga gagggcggag aggaggagga 8940

tcctgttgga gagcgacttg gaccctggca gctgaaccgc cccggagatc tccctgatgg 9000

gctggagcac gatctcctcc gccttcgccg ccggcgctgc caccgacgac gacgacgcgg 9060

acgccaccac caccgcctcc cgccgccccc gcgcccgcac ccgcacccgc atccccccgc 9120

gcgccgcggc gggcagccgc agctgcttcc gcgacgagaa cgccgccgac gccgccacgg 9180

cctggtccag ggacaccgcc gccgcagccg cggcgttgga cgccatggtc gccgccattc 9240

tagagcggcc gctctagaac tagtggatcc gaagtaacac caaacaacag ggtgagcatc 9300

gacaaaagaa acagtaccaa gcaaataaat agcgtatgaa ggcagggcta aaaaaatcca 9360

catatagctg ctgcatatgc catcatccaa gtatatcaag atcaaaataa ttataaaaca 9420

tacttgttta ttataataga taggtactca aggttagagc atatgaatag atgctgcata 9480

tgccatcatg tatatgcatc agtaaaaccc acatcaacat gtatacctat cctagatcga 9540

tatttccatc catcttaaac tcgtaactat gaagatgtat gacacacaca tacagttcca 9600

aaattaataa atacaccagg tagtttgaaa cagtattcta ctccgatcta gaacgaatga 9660

acgaccgccc aaccacacca catcatcaca accaagcgaa caaaaagcat ctctgtatat 9720

gcatcagtaa aacccgcatc aacatgtata cctatcctag atcgatattt ccatccatca 9780

tcttcaattc gtaactatga atatgtatgg cacacacata cagatccaaa attaataaat 9840

ccaccaggta gtttgaaaca gaattctact ccgatctaga acgaccgccc aaccagacca 9900

catcatcaca accaagacaa aaaaaagcat gaaaagatga cccgacaaac aagtgcacgg 9960

catatattga aataaaggaa aagggcaaac caaaccctat gcaacgaaac aaaaaaaatc 10020

atgaaatcga tcccgtctgc ggaacggcta gagccatccc aggattcccc aaagagaaac 10080

actggcaagt tagcaatcag aacgtgtctg acgtacaggt cgcatccgtg tacgaacgct 10140

agcagcacgg atctaacaca aacacggatc taacacaaac atgaacagaa gtagaactac 10200

cgggccctaa ccatggaccg gaacgccgat ctagagaagg tagagagggg ggggggggga 10260

ggacgagcgg cgtaccttga agcggaggtg ccgacgggtg gatttggggg agatctggtt 10320

gtgtgtgtgt gcgctccgaa caacacgagg ttggggaaag agggtgtgga gggggtgtct 10380

atttattacg gcgggcgagg aagggaaagc gaaggagcgg tgggaaagga atcccccgta 10440

gctgccgtgc cgtgagagga ggaggaggcc gcctgccgtg ccggctcacg tctgccgctc 10500

cgccacgcaa tttctggatg ccgacagcgg agcaagtcca acggtggagc ggaactctcg 10560

agaggggtcc agaggcagcg acagagatgc cgtgccgtct gcttcgcttg gcccgacgcg 10620

acgctgctgg ttcgctggtt ggtgtccgtt agactcgtcg acggcgttta acaggctggc 10680

attatctact cgaaacaaga aaaatgtttc cttagttttt ttaatttctt aaagggtatt 10740

tgtttaattt ttagtcactt tattttattc tattttatat ctaaattatt aaataaaaaa 10800

actaaaatag agttttagtt ttcttaattt agaggctaaa atagaataaa atagatgtac 10860

taaaaaaatt agtctataaa aaccattaac cctaaaccct aaatggatgt actaataaaa 10920

tggatgaagt attatatagg tgaagctatt tgcaaaaaaa aaggagaaca catgcacact 10980

aaaaagataa aactgtagag tcctgttgtc aaaatactca attgtccttt agaccatgtc 11040

taactgttca tttatatgat tctctaaaac actgatatta ttgtagtact atagattata 11100

ttattcgtag agtaaagttt aaatatatgt ataaagatag ataaactgca cttcaacaag 11160

cttggcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact 11220

taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac 11280

cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgctagagca gcttgagctt 11340

ggatcagatt gtcgtttccc gccttcagtt tggggatcct ctagagtcga cctgcaggca 11400

tgcaagcttg gcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac 11460

ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc 11520

ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatgct agagcagctt 11580

gagcttggat cagattgtcg tttcccgcct tcagtttaat tcgatccatg cctcacatgt 11640

taatgtacta ccaatggagg gctgtacaca tttatgatta cgaaattttt taatatattt 11700

tatagatttc ttatgcatca tacaaaaata cataattatt cgtaacattt tggagataca 11760

tattcagatg catcaaattc taattaaacg ttaaaatatt ttggagacgt atcttcgtaa 11820

caatttaaaa cctatactat acatcacatt cgaaggtcat tttataattt aaaatattat 11880

ggagatgcat cttcgtttat gtttgctcag atgaagattt aaaccttaca aacaatatgt 11940

aaaaaatgac gtacataaat tcagatagtc caaaagtgtc atatataaat aaagatcaat 12000

aagtgtcaaa aaaagtcaag aacaacgata gagtagcata atgtcaaaat aaaataaaat 12060

ccatgacact actactatta tatattaatg cactaatgtg tatgtctaac tacatcgtct 12120

ctgcctcctc tgtcagttat gtctcgtaag ccatcaatcc cccgtcctcc ggcgttgtct 12180

ccggtatatc aatgtcccca tgtgcctacg tcatgatggc atctaggaca tgtctcacat 12240

cagacacatt aggaaagata catttgccaa tgtatatctg cgcaatctcc acaatgcaac 12300

gacatatagg caagacatcc tcaacataat ttagttgtgc atgcttctcc tctagtatct 12360

cccgatgagt tgatcaagct tatcgaaact atcagtgttt gacaggatat attggcgggt 12420

aaacctaaga gaaaagagcg tttattagaa taacggatat ttaaaagggc gtgaaaaggt 12480

ttatccgttc gtccatttgt atgtg 12505

<210> 21

<211> 28

<212> DNA

<213> 人工序列(Artificial sequence)

<400> 21

cgagctcgga tctagtaaca tagatgac 28

<210> 22

<211> 30

<212> DNA

<213> 人工序列(Artificial sequence)

<400> 22

ggggtacccc gagctcgaat ttccccgatc 30

Claims (8)

1.一种多功能抗草甘膦水稻转化载体pCDMAR-epsps，其特征在于，所述载体pCDMAR-epsps的大小为12774bp，序列如SEQ ID NO：1所示；且所述载体pCDMAR-epsps中包括11个功能元件，具体见下表：

2.一种多功能抗草甘膦水稻转化载体pCDMAR-epsps的构建方法，其特征在于，所述构建方法包括以下步骤：

(1)骨架选择：选择pCDMAR-hyg作为骨架，且所述pCDMAR-hyg骨架大小为11724bp，序列如SEQ ID NO：14所示；

(2)含pCDMAR-hyg骨架的感受态细胞制备及培养：将pCDMAR-hyg骨架转化到感受态细胞中，并采用常规方法进行培养，获得含pCDMAR-hyg骨架的感受态细胞；

(3)酶切：采用BclI酶酶切所述pCDMAR-hyg骨架，得到片段Ⅰ、片段Ⅱ、片段Ⅲ；且所述片段Ⅰ的大小为6067bp，序列如SEQ ID NO：15所示；片段Ⅱ的大小为2975bp，序列如SEQ IDNO：16所示；片段Ⅲ的大小为2682bp，序列如SEQ ID NO：17所示；

(4)连接：将所述片段Ⅰ、片段Ⅱ连接得到连接产物，并通过ClaI/EcoRV酶切鉴定所述连接产物；选择ClaI/EcoRV酶切后片段大小为7346bp和1403bp的连接产物，并将其命名为中间质粒pCDMAR；所述中间质粒pCDMAR的大小为8749bp，序列如SEQ ID NO：18所示；

(5)pCDMAR-UbiEP中间产物的获得：利用SbfI/KpnI酶酶切pCUEP102质粒获得酶切产物，并回收其中大小为3783bp的片段，命名为酶切片段Ⅰ，序列如SEQ ID NO：19所示；之后利用SbfI/KpnI酶切所述中间质粒pCDMAR，回收酶切片段，命名为酶切片段Ⅱ；之后将所述酶切片段Ⅰ和酶切片段Ⅱ进行连接，获得连接产物，命名为pCDMAR-UbiEP中间产物；所述pCDMAR-UbiEP中间产物的大小为12505bp，序列如SEQ ID NO：20所示；

(6)带有酶切位点的T-nos片段的获得：设计两端加KpnI和SacI酶切位点的引物，并采用所述引物扩增pCUEP102上T-nos片段，回收大小为269bp的目的条带，且所述目的条带的序列如SEQ ID NO：8所示；

(7)酶切连接：将所述目的条带与所述pCDMAR-UbiEP中间产物通过KpnI/SacI酶进行酶切连接，得到连接产物，即为一种多功能抗草甘膦水稻转化载体pCDMAR-epsps；且所述载体pCDMAR-epsps的大小为12774bp，序列如SEQ ID NO：1所示。

3.根据权利要求2所述的一种多功能抗草甘膦水稻转化载体pCDMAR-epsps的构建方法，其特征在于，步骤(2)中所述感受态细胞为Trans110感受态细胞。

4.根据权利要求2所述的一种多功能抗草甘膦水稻转化载体pCDMAR-epsps的构建方法，其特征在于，步骤(4)中所述片段Ⅰ、片段Ⅱ两条片段连接过程使用的连接酶为T4连接酶。

5.根据权利要求2所述的一种多功能抗草甘膦水稻转化载体pCDMAR-epsps的构建方法，其特征在于，步骤(7)中所述目的条带与所述pCDMAR-UbiEP中间产物通过KpnI/SacI酶进行酶切连接的连接过程使用的连接酶为T4连接酶。

6.根据权利要求2所述的一种多功能抗草甘膦水稻转化载体pCDMAR-epsps的构建方法，其特征在于，步骤(6)中所述引物包括上游引物和下游引物，且所述上游引物序列如SEQID NO：21所示，所述下游引物序列如SEQ ID NO：22所示。

7.根据权利要求1所述的一种多功能抗草甘膦水稻转化载体pCDMAR-epsps在获得抗除草剂的水稻品种的应用。

8.根据权利要求1所述的一种多功能抗草甘膦水稻转化载体pCDMAR-epsps在农业领域的应用。

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