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基本信息

體類(lèi)型 

大腸桿菌蛋白雙表達(dá)載體,有兩個(gè)多克隆位點(diǎn)(雙啟動(dòng)子) 

載體抗性 

Kan 

載體標(biāo)簽 

N-His, C-S 

篩選標(biāo)記 

/ 

啟動(dòng)子 

T7lac 

表達(dá)水平 

高拷貝 

載體大小 

3829bp 

克隆方法 

多克隆位點(diǎn),限制性?xún)?nèi)切酶 

5' 測(cè)序引物 

DUETF1: TGGTGTCCGGGATCTCGA 

DUETF2: TTGTACACGGCCGCATAATC 

3' 測(cè)序引物 

DUETR1: 5'-GATTATGCGGCCGTGTACAA-3'  

T7 ter: TGCTAGTTATTGCTCAGCGG 

體類(lèi)型 

大腸桿菌蛋白雙表達(dá)載體,有兩個(gè)多克隆位點(diǎn)(雙啟動(dòng)子) 

備注:

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質(zhì)粒圖譜和多克隆位點(diǎn)信息

注意:

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載體描述

pRSFDuet-1 is designed for the coexpression of two target ORFs. The vector contains two multiple cloning sites (MCS), each of which is preceded by a T7lac promoter and ribosome binding site (rbs). The vector also carries the RSF1030-derived RSF replicon, lacI gene and kanamycin resistance gene (KnR). This vector can be used in combination with pACYCDuet?-1, pCDFDuet?-1, and pETDuet?-1 in an appropriate host strain for the coexpression of 4 to 8 target proteins. ORFs inserted into MCS1 can be sequenced using the ACYCDuetUP1 Primer and DuetDOWN1 Primer. ORFs inserted into MCS2 can be sequenced using the DuetUP2 Primer and T7 Terminator Primer. Unique sites are shown on the circle map.

載體序列

ORIGIN

        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 catgcagcgc tcttccgctt cctcgctcac

     1681 tgactcgcta cgctcggtcg ttcgactgcg gcgagcggtg tcagctcact caaaagcggt

     1741 aatacggtta tccacagaat caggggataa agccggaaag aacatgtgag caaaaagcaa

     1801 agcaccggaa gaagccaacg ccgcaggcgt ttttccatag gctccgcccc cctgacgagc

     1861 atcacaaaaa tcgacgctca agccagaggt ggcgaaaccc gacaggacta taaagatacc

     1921 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg

     1981 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgtt

     2041 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg

     2101 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac

     2161 acgacttatc gccactggca gcagccattg gtaactgatt tagaggactt tgtcttgaag

     2221 ttatgcacct gttaaggcta aactgaaaga acagattttg gtgagtgcgg tcctccaacc

     2281 cacttacctt ggttcaaaga gttggtagct cagcgaacct tgagaaaacc accgttggta

     2341 gcggtggttt ttctttattt atgagatgat gaatcaatcg gtctatcaag tcaacgaaca

     2401 gctattccgt tactctagat ttcagtgcaa tttatctctt caaatgtagc acctgaagtc

     2461 agccccatac gatataagtt gtaattctca tgttagtcat gccccgcgcc caccggaagg

     2521 agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga

     2581 gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt

     2641 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc

     2701 agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg

     2761 ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt

     2821 ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact

     2881 accgagatgt ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc

     2941 gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc

     3001 atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga

     3061 atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa

     3121 cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg

     3181 cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag

     3241 acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg

     3301 tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc

     3361 gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc

     3421 agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga

     3481 ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg

     3541 ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa

     3601 acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct

     3661 gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg

     3721 cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg gatctcgacg

     3781 ctctccctta tgcgactcct gcattaggaa attaatacga ctcactata

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