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基本信息

質(zhì)粒圖譜和多克隆位點(diǎn)信息 

注意：

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載體描述

pCOLADuet-1 is designed for the coexpression of two target genes from a single plasmid. The vector encodes two multiple cloning sites (MCS) each of which is preceded by a T7 promoter, lac operator, and ribosome binding site (rbs). MCS-1 encodes the six-amino acid His?Tag sequence for the creation of a N-terminal fusion and MCS2 encodes the 15 amino acid S?Tag? peptide after the last restriction site for the creation of a C-terminal fusion if desired. Genes inserted into MCS-1 can be sequenced using the ACYCDuetUP1 Primer and DuetDOWN1 Primer. Genes inserted into MCS-2 can be sequenced using the DuetUP2 Primer and T7 Terminator Primer. The vector has the COLA replicon from ColA(1) and the kanamycin resistance gene. This vector can be transformed into the same cell with plasmids containing compatible origins of replication and drug resistance genes for coexpression of up to 8 target genes.

載體序列

         1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag

       61 gagatatacc atgggcagca gccatcacca tcatcaccac agccaggatc cgaattcgag

      121 ctcggcgcgc ctgcaggtcg acaagcttgc ggccgcataa tgcttaagtc gaacagaaag

      181 taatcgtatt gtacacggcc gcataatcga aattaatacg actcactata ggggaattgt

      241 gagcggataa caattcccca tcttagtata ttagttaagt ataagaagga gatatacata

      301 tggcagatct caattggata tcggccggcc acgcgatcgc tgacgtcggt accctcgagt

      361 ctggtaaaga aaccgctgct gcgaaatttg aacgccagca catggactcg tctactagcg

      421 cagcttaatt aacctaggct gctgccaccg ctgagcaata actagcataa ccccttgggg

      481 cctctaaacg ggtcttgagg ggttttttgc tgaaacctca ggcatttgag aagcacacgg

      541 tcacactgct tccggtagtc aataaaccgg taaaccagca atagacataa gcggctattt

      601 aacgaccctg ccctgaaccg acgacaagct gacgaccggg tctccgcaag tggcactttt

      661 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat

      721 ccgctcatga attaattctt agaaaaactc atcgagcatc aaatgaaact gcaatttatt

      781 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa

      841 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg

      901 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa

      961 atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca

     1021 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc

     1081 gttattcatt cgtgattgcg cctgagcgag acgaaatacg cggtcgctgt taaaaggaca

     1141 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt

     1201 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt

     1261 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat

     1321 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc

     1381 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt

     1441 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat

     1501 gttggaattt aatcgcggcc tagagcaaga cgtttcccgt tgaatatggc tcatactctt

     1561 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt

     1621 tgaatgtatt tagaaaaata aacaaatagg catgctagcg cagaaacgtc ctagaagatg

     1681 ccaggaggat acttagcaga gagacaataa ggccggagcg aagccgtttt tccataggct

     1741 ccgcccccct gacgaacatc acgaaatctg acgctcaaat cagtggtggc gaaacccgac

     1801 aggactataa agataccagg cgtttccccc tgatggctcc ctcttgcgct ctcctgttcc

     1861 cgtcctgcgg cgtccgtgtt gtggtggagg ctttacccaa atcaccacgt cccgttccgt

     1921 gtagacagtt cgctccaagc tgggctgtgt gcaagaaccc cccgttcagc ccgactgctg

     1981 cgccttatcc ggtaactatc atcttgagtc caacccggaa agacacgaca aaacgccact

     2041 ggcagcagcc attggtaact gagaattagt ggatttagat atcgagagtc ttgaagtggt

     2101 ggcctaacag aggctacact gaaaggacag tatttggtat ctgcgctcca ctaaagccag

     2161 ttaccaggtt aagcagttcc ccaactgact taaccttcga tcaaaccgcc tccccaggcg

     2221 gttttttcgt ttacagagca ggagattacg acgatcgtaa aaggatctca agaagatcct

     2281 ttacggattc ccgacaccat cactctagat ttcagtgcaa tttatctctt caaatgtagc

     2341 acctgaagtc agccccatac gatataagtt gtaattctca tgttagtcat gccccgcgcc

     2401 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc

     2461 taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga

     2521 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt

     2581 attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt

     2641 caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg

     2701 aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc

     2761 gtatcccact accgagatgt ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat

     2821 tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt

     2881 cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc

     2941 tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc

     3001 cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag

     3061 atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt

     3121 ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat

     3181 ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag

     3241 attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac

     3301 gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg

     3361 cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg

     3421 tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt

     3481 tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc

     3541 ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact

     3601 ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg

     3661 gatctcgacg ctctccctta tgcgactcct gcattaggaa attaatacga ctcactata

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