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基本信息
载体类型: | 大肠杆菌蛋白表达载体,低温诱导表达载体 |
载体抗性: | Amp |
载体标签: | N‐His, N‐Factor Xa,N-ZZ domain |
筛选标记: | / |
启动子: | cspA |
表达水平: | 高 |
载体大小: | 4788bp |
克隆方法: | 多克隆位点,限制性内切酶 |
5' 测序引物: | pCold‐F:5'‐ACGCCATATCGCCGAAAGG‐3' |
3' 测序引物: | pCold‐R:5'‐GGCAGGGATCTTAGATTCTG‐3' |
备注: | 如有侵权请产权持有人尽快联系法务15927229196 |
载体描述
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由质粒pCold-I改造,在Ndel与Sacl酶切位点间,插入ZZdomain:
Z Zdomain:
GACAACAAATTCAACAAAGAACAACAAAACGCGTTCTATGAGATCTTACATTTACCTAACTTAAACGAAGAACAACGAAACGCCTTCATCCAAAGTTTAAAAGATGACCCAAGCCAAAGCGCTAACCTTTTAGCAGAAGCTAAAAAGCTAAATGATGCTCAGGCGCCGAAAGTAGACAACAAATTCAACAAAGAACAACAAAACGCGTTCTATGAGATCTTACATTTACCTAACTTAAACGAAGAACAACGAAACGCCTTCATCCAAAGTTTAAAAGATGACCCAAGCCAAAGCGCTAACCTTTTAGCAGAAGCTAAAAAGCTAAATGATGCTCAGGCGCCGAAAGTA
ZZ domain总长348bp,由两段完全一样的序列组成,改造后为TEE+His+Factor Xa+ZZ+多克隆位点。
载体序列
ORIGIN
1 aaggaatggt gtggccgatt aatcataaat atgaaaaata attgttgcat cacccgccaa
61 tgcgtggctt aatgcacatc aaattgtgag cggataacaa tttgatgtgc tagcgcatat
121 ccagtgtagt aaggcaagtc ccttcaagag ttatcgttga tacccctcgt agtgcacatt
181 cctttaacgc ttcaaaatct gtaaagcacg ccatatcgcc gaaaggcaca cttaattatt
241 aagaggtaat acaccatgaa tcacaaagtg catcatcatc atcatcatat cgaaggtagg
301 catatggcgc aacacgatga agccgtagac aacaaattca acaaagaaca acaaaacgcg
361 ttctatgaga tcttacattt acctaactta aacgaagaac aacgaaacgc cttcatccaa
421 agtttaaaag atgacccaag ccaaagcgct aaccttttag cagaagctaa aaagctaaat
481 gatgctcagg cgccgaaagt agacaacaaa ttcaacaaag aacaacaaaa cgcgttctat
541 gagatcttac atttacctaa cttaaacgaa gaacaacgaa acgccttcat ccaaagttta
601 aaagatgacc caagccaaag cgctaacctt ttagcagaag ctaaaaagct aaatgatgct
661 caggcgccga aagtagacgc gacattcgag ctcggtaccc tcgagggatc cgaattcaag
721 cttgtcgacc tgcagtctag ataggtaatc tctgcttaaa agcacagaat ctaagatccc
781 tgccatttgg cggggatttt tttatttgtt ttcaggaaat aaataatcga tcgcgtaata
841 aaatctatta ttatttttgt gaagaataaa tttgggtgca atgagaatgc gcaggccctt
901 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
961 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
1021 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
1081 agtgcaccat aaaattgtaa acgttaatat tttgttaaaa ttcgcgttaa atttttgtta
1141 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
1201 atagcccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
1261 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
1321 accatcaccc aaatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
1381 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
1441 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
1501 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtact atggttgctt
1561 tgacgtatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgt
1621 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
1681 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
1741 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
1801 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
1861 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
1921 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
1981 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
2041 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
2101 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
2161 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
2221 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
2281 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
2341 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
2401 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
2461 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
2521 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
2581 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
2641 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
2701 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
2761 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
2821 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
2881 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
2941 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
3001 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
3061 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
3121 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
3181 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
3241 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
3301 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
3361 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
3421 ttgctcacat agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca
3481 agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc
3541 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
3601 aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt
3661 gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg
3721 tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata
3781 taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatatccgc accaacgcgc
3841 agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc
3901 atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg
3961 gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta
4021 tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg
4081 atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg
4141 gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca
4201 ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc
4261 agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg
4321 cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc
4381 gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc
4441 aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc
4501 atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg
4561 cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt
4621 ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag
4681 gttttgcgcc attcgatggt gtccgggatc tcgacgctct cccttatgcg actcctgcat
4741 taggaagcag cccagtagta ggttgaggcc gttgagcacc gccgccgc
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