pTrcHis2 C

pTrcHis2 C

pTrcHis2C载体基本信息

pTrcHis2C载体质粒图谱和多克隆位点信息

pTrcHis2C载体简介

The pTrcHis2 plasmids are pBR322-derived expression vectors designed for efficient recombinant protein expression and purification in E. coli. High levels of expression are possible using the trc (trp-lac) promoter (Egon et al., 1983) and the rrnB anti-termination region (Li et al., 1984). The trc promoter contains the –35 region of the trp promoter together with the –10 region of the lac promoter (Brosius et al., 1985; Egon et al., 1983; Mulligan et al., 1985). To regulate expression, the gene encoding Lac repressor (lacIq) is provided in the pTrcHis2 vectors, allowing regulation of the trc promoter regardless of whether the host strain contains a gene encoding the Lac repressor.

Isopropyl-Beta-D-thiogalactopyranoside (IPTG) is used to induce expression of your gene. Translation is enhanced by the bacteriophage T7 gene 10 translation enhancer and a minicistron that provides highly efficient translational restart into the open reading frame of the multiple cloning site. DNA inserts are positioned downstream and in frame with the initiation ATG and a C-terminal fusion peptide. The C-terminal peptide encodes the myc epitope and six histidine residues that function as a metal binding site in the expressed protein.

pTrcHis2C载体序列

ORIGIN
        1 GTTTGACAGC TTATCATCGA CTGCACGGTG CACCAATGCT TCTGGCGTCA GGCAGCCATC
       61 GGAAGCTGTG GTATGGCTGT GCAGGTCGTA AATCACTGCA TAATTCGTGT CGCTCAAGGC
      121 GCACTCCCGT TCTGGATAAT GTTTTTTGCG CCGACATCAT AACGGTTCTG GCAAATATTC
      181 TGAAATGAGC TGTTGACAAT TAATCATCCG GCTCGTATAA TGTGTGGAAT TGTGAGCGGA
      241 TAACAATTTC ACACAGGAAA CAGCGCCGCT GAGAAAAAGC GAAGCGGCAC TGCTCTTTAA
      301 CAATTTATCA GACAATCTGT GTGGGCACTC GACCGGAATT ATCGATTAAC TTTATTATTA
      361 AAAATTAAAG AGGTATATAT TAATGTATCG ATTAAATAAG GAGGAATAAA CCATGGATCC
      421 GAGCTCGAGA TCTGCAGCTG GTACCATATG GGAATTCGAA GCTTACGTAG AACAAAAACT
      481 CATCTCAGAA GAGGATCTGA ATAGCGCCGT CGACCATCAT CATCATCATC ATTGAGTTTA
      541 AACGGTCTCC AGCTTGGCTG TTTTGGCGGA TGAGAGAAGA TTTTCAGCCT GATACAGATT
      601 AAATCAGAAC GCAGAAGCGG TCTGATAAAA CAGAATTTGC CTGGCGGCAG TAGCGCGGTG
      661 GTCCCACCTG ACCCCATGCC GAACTCAGAA GTGAAACGCC GTAGCGCCGA TGGTAGTGTG
      721 GGGTCTCCCC ATGCGAGAGT AGGGAACTGC CAGGCATCAA ATAAAACGAA AGGCTCAGTC
      781 GAAAGACTGG GCCTTTCGTT TTATCTGTTG TTTGTCGGTG AACGCTCTCC TGAGTAGGAC
      841 AAATCCGCCG GGAGCGGATT TGAACGTTGC GAAGCAACGG CCCGGAGGGT GGCGGGCAGG
      901 ACGCCCGCCA TAAACTGCCA GGCATCAAAT TAAGCAGAAG GCCATCCTGA CGGATGGCCT
      961 TTTTGCGTTT CTACAAACTC TTTTTGTTTA TTTTTCTAAA TACATTCAAA TATGTATCCG
     1021 CTCATGAGAC AATAACCCTG ATAAATGCTT CAATAATATT GAAAAAGGAA GAGTATGAGT
     1081 ATTCAACATT TCCGTGTCGC CCTTATTCCC TTTTTTGCGG CATTTTGCCT TCCTGTTTTT
     1141 GCTCACCCAG AAACGCTGGT GAAAGTAAAA GATGCTGAAG ATCAGTTGGG TGCACGAGTG
     1201 GGTTACATCG AACTGGATCT CAACAGCGGT AAGATCCTTG AGAGTTTTCG CCCCGAAGAA
     1261 CGTTTTCCAA TGATGAGCAC TTTTAAAGTT CTGCTATGTG GCGCGGTATT ATCCCGTGTT
     1321 GACGCCGGGC AAGAGCAACT CGGTCGCCGC ATACACTATT CTCAGAATGA CTTGGTTGAG
     1381 TACTCACCAG TCACAGAAAA GCATCTTACG GATGGCATGA CAGTAAGAGA ATTATGCAGT
     1441 GCTGCCATAA CCATGAGTGA TAACACTGCG GCCAACTTAC TTCTGACAAC GATCGGAGGA
     1501 CCGAAGGAGC TAACCGCTTT TTTGCACAAC ATGGGGGATC ATGTAACTCG CCTTGATCGT
     1561 TGGGAACCGG AGCTGAATGA AGCCATACCA AACGACGAGC GTGACACCAC GATGCCTGTA
     1621 GCAATGGCAA CAACGTTGCG CAAACTATTA ACTGGCGAAC TACTTACTCT AGCTTCCCGG
     1681 CAACAATTAA TAGACTGGAT GGAGGCGGAT AAAGTTGCAG GACCACTTCT GCGCTCGGCC
     1741 CTTCCGGCTG GCTGGTTTAT TGCTGATAAA TCTGGAGCCG GTGAGCGTGG GTCTCGCGGT
     1801 ATCATTGCAG CACTGGGGCC AGATGGTAAG CCCTCCCGTA TCGTAGTTAT CTACACGACG
     1861 GGGAGTCAGG CAACTATGGA TGAACGAAAT AGACAGATCG CTGAGATAGG TGCCTCACTG
     1921 ATTAAGCATT GGTAACTGTC AGACCAAGTT TACTCATATA TACTTTAGAT TGATTTAAAA
     1981 CTTCATTTTT AATTTAAAAG GATCTAGGTG AAGATCCTTT TTGATAATCT CATGACCAAA
     2041 ATCCCTTAAC GTGAGTTTTC GTTCCACTGA GCGTCAGACC CCGTAGAAAA GATCAAAGGA
     2101 TCTTCTTGAG ATCCTTTTTT TCTGCGCGTA ATCTGCTGCT TGCAAACAAA AAAACCACCG
     2161 CTACCAGCGG TGGTTTGTTT GCCGGATCAA GAGCTACCAA CTCTTTTTCC GAAGGTAACT
     2221 GGCTTCAGCA GAGCGCAGAT ACCAAATACT GTCCTTCTAG TGTAGCCGTA GTTAGGCCAC
     2281 CACTTCAAGA ACTCTGTAGC ACCGCCTACA TACCTCGCTC TGCTAATCCT GTTACCAGTG
     2341 GCTGCTGCCA GTGGCGATAA GTCGTGTCTT ACCGGGTTGG ACTCAAGACG ATAGTTACCG
     2401 GATAAGGCGC AGCGGTCGGG CTGAACGGGG GGTTCGTGCA CACAGCCCAG CTTGGAGCGA
     2461 ACGACCTACA CCGAACTGAG ATACCTACAG CGTGAGCTAT GAGAAAGCGC CACGCTTCCC
     2521 GAAGGGAGAA AGGCGGACAG GTATCCGGTA AGCGGCAGGG TCGGAACAGG AGAGCGCACG
     2581 AGGGAGCTTC CAGGGGGAAA CGCCTGGTAT CTTTATAGTC CTGTCGGGTT TCGCCACCTC
     2641 TGACTTGAGC GTCGATTTTT GTGATGCTCG TCAGGGGGGC GGAGCCTATG GAAAAACGCC
     2701 AGCAACGCGG CCTTTTTACG GTTCCTGGCC TTTTGCTGGC CTTTTGCTCA CATGTTCTTT
     2761 CCTGCGTTAT CCCCTGATTC TGTGGATAAC CGTATTACCG CCTTTGAGTG AGCTGATACC
     2821 GCTCGCCGCA GCCGAACGAC CGAGCGCAGC GAGTCAGTGA GCGAGGAAGC GGAAGAGCGC
     2881 CTGATGCGGT ATTTTCTCCT TACGCATCTG TGCGGTATTT CACACCGCAT ATGGTGCACT
     2941 CTCAGTACAA TCTGCTCTGA TGCCGCATAG TTAAGCCAGT ATACACTCCG CTATCGCTAC
     3001 GTGACTGGGT CATGGCTGCG CCCCGACACC CGCCAACACC CGCTGACGCG CCCTGACGGG
     3061 CTTGTCTGCT CCCGGCATCC GCTTACAGAC AAGCTGTGAC CGTCTCCGGG AGCTGCATGT
     3121 GTCAGAGGTT TTCACCGTCA TCACCGAAAC GCGCGAGGCA GCAGATCAAT TCGCGCGCGA
     3181 AGGCGAAGCG GCATGCATTT ACGTTGACAC CATCGAATGG TGCAAAACCT TTCGCGGTAT
     3241 GGCATGATAG CGCCCGGAAG AGAGTCAATT CAGGGTGGTG AATGTGAAAC CAGTAACGTT
     3301 ATACGATGTC GCAGAGTATG CCGGTGTCTC TTATCAGACC GTTTCCCGCG TGGTGAACCA
     3361 GGCCAGCCAC GTTTCTGCGA AAACGCGGGA AAAAGTGGAA GCGGCGATGG CGGAGCTGAA
     3421 TTACATTCCC AACCGCGTGG CACAACAACT GGCGGGCAAA CAGTCGTTGC TGATTGGCGT
     3481 TGCCACCTCC AGTCTGGCCC TGCACGCGCC GTCGCAAATT GTCGCGGCGA TTAAATCTCG
     3541 CGCCGATCAA CTGGGTGCCA GCGTGGTGGT GTCGATGGTA GAACGAAGCG GCGTCGAAGC
     3601 CTGTAAAGCG GCGGTGCACA ATCTTCTCGC GCAACGCGTC AGTGGGCTGA TCATTAACTA
     3661 TCCGCTGGAT GACCAGGATG CCATTGCTGT GGAAGCTGCC TGCACTAATG TTCCGGCGTT
     3721 ATTTCTTGAT GTCTCTGACC AGACACCCAT CAACAGTATT ATTTTCTCCC ATGAAGACGG
     3781 TACGCGACTG GGCGTGGAGC ATCTGGTCGC ATTGGGTCAC CAGCAAATCG CGCTGTTAGC
     3841 GGGCCCATTA AGTTCTGTCT CGGCGCGTCT GCGTCTGGCT GGCTGGCATA AATATCTCAC
     3901 TCGCAATCAA ATTCAGCCGA TAGCGGAACG GGAAGGCGAC TGGAGTGCCA TGTCCGGTTT
     3961 TCAACAAACC ATGCAAATGC TGAATGAGGG CATCGTTCCC ACTGCGATGC TGGTTGCCAA
     4021 CGATCAGATG GCGCTGGGCG CAATGCGCGC CATTACCGAG TCCGGGCTGC GCGTTGGTGC
     4081 GGATATCTCG GTAGTGGGAT ACGACGATAC CGAAGACAGC TCATGTTATA TCCCGCCGTC
     4141 AACCACCATC AAACAGGATT TTCGCCTGCT GGGGCAAACC AGCGTGGACC GCTTGCTGCA
     4201 ACTCTCTCAG GGCCAGGCGG TGAAGGGCAA TCAGCTGTTG CCCGTCTCAC TGGTGAAAAG
     4261 AAAAACCACC CTGGCGCCCA ATACGCAAAC CGCCTCTCCC CGCGCGTTGG CCGATTCATT
     4321 AATGCAGCTG GCACGACAGG TTTCCCGACT GGAAAGCGGG CAGTGAGCGC AACGCAATTA
     4381 ATGTGAGTTA GCGCGAATTG ATCTG
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