pTrcHis2 A

pTrcHis2A载体基本信息

pTrcHis2A载体质粒图谱和多克隆位点信息

pTrcHis2A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.

pTrcHis2A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 GCTTGGGCCC GAACAAAAAC

  481 TCATCTCAGA AGAGGATCTG AATAGCGCCG TCGACCATCA TCATCATCAT CATTGAGTTT

  541 AAACGGTCTC CAGCTTGGCT GTTTTGGCGG ATGAGAGAAG ATTTTCAGCC TGATACAGAT

  601 TAAATCAGAA CGCAGAAGCG GTCTGATAAA ACAGAATTTG CCTGGCGGCA GTAGCGCGGT

  661 GGTCCCACCT GACCCCATGC CGAACTCAGA AGTGAAACGC CGTAGCGCCG ATGGTAGTGT

  721 GGGGTCTCCC CATGCGAGAG TAGGGAACTG CCAGGCATCA AATAAAACGA AAGGCTCAGT

  781 CGAAAGACTG GGCCTTTCGT TTTATCTGTT GTTTGTCGGT GAACGCTCTC CTGAGTAGGA

  841 CAAATCCGCC GGGAGCGGAT TTGAACGTTG CGAAGCAACG GCCCGGAGGG TGGCGGGCAG

  901 GACGCCCGCC ATAAACTGCC AGGCATCAAA TTAAGCAGAA GGCCATCCTG ACGGATGGCC

  961 TTTTTGCGTT TCTACAAACT CTTTTTGTTT ATTTTTCTAA ATACATTCAA ATATGTATCC

 1021 GCTCATGAGA CAATAACCCT GATAAATGCT TCAATAATAT TGAAAAAGGA AGAGTATGAG

 1081 TATTCAACAT TTCCGTGTCG CCCTTATTCC CTTTTTTGCG GCATTTTGCC TTCCTGTTTT

 1141 TGCTCACCCA GAAACGCTGG TGAAAGTAAA AGATGCTGAA GATCAGTTGG GTGCACGAGT

 1201 GGGTTACATC GAACTGGATC TCAACAGCGG TAAGATCCTT GAGAGTTTTC GCCCCGAAGA

 1261 ACGTTTTCCA ATGATGAGCA CTTTTAAAGT TCTGCTATGT GGCGCGGTAT TATCCCGTGT

 1321 TGACGCCGGG CAAGAGCAAC TCGGTCGCCG CATACACTAT TCTCAGAATG ACTTGGTTGA

 1381 GTACTCACCA GTCACAGAAA AGCATCTTAC GGATGGCATG ACAGTAAGAG AATTATGCAG

 1441 TGCTGCCATA ACCATGAGTG ATAACACTGC GGCCAACTTA CTTCTGACAA CGATCGGAGG

 1501 ACCGAAGGAG CTAACCGCTT TTTTGCACAA CATGGGGGAT CATGTAACTC GCCTTGATCG

 1561 TTGGGAACCG GAGCTGAATG AAGCCATACC AAACGACGAG CGTGACACCA CGATGCCTGT

 1621 AGCAATGGCA ACAACGTTGC GCAAACTATT AACTGGCGAA CTACTTACTC TAGCTTCCCG

 1681 GCAACAATTA ATAGACTGGA TGGAGGCGGA TAAAGTTGCA GGACCACTTC TGCGCTCGGC

 1741 CCTTCCGGCT GGCTGGTTTA TTGCTGATAA ATCTGGAGCC GGTGAGCGTG GGTCTCGCGG

 1801 TATCATTGCA GCACTGGGGC CAGATGGTAA GCCCTCCCGT ATCGTAGTTA TCTACACGAC

 1861 GGGGAGTCAG GCAACTATGG ATGAACGAAA TAGACAGATC GCTGAGATAG GTGCCTCACT

 1921 GATTAAGCAT TGGTAACTGT CAGACCAAGT TTACTCATAT ATACTTTAGA TTGATTTAAA

 1981 ACTTCATTTT TAATTTAAAA GGATCTAGGT GAAGATCCTT TTTGATAATC TCATGACCAA

 2041 AATCCCTTAA CGTGAGTTTT CGTTCCACTG AGCGTCAGAC CCCGTAGAAA AGATCAAAGG

 2101 ATCTTCTTGA GATCCTTTTT TTCTGCGCGT AATCTGCTGC TTGCAAACAA AAAAACCACC

 2161 GCTACCAGCG GTGGTTTGTT TGCCGGATCA AGAGCTACCA ACTCTTTTTC CGAAGGTAAC

 2221 TGGCTTCAGC AGAGCGCAGA TACCAAATAC TGTCCTTCTA GTGTAGCCGT AGTTAGGCCA

 2281 CCACTTCAAG AACTCTGTAG CACCGCCTAC ATACCTCGCT CTGCTAATCC TGTTACCAGT

 2341 GGCTGCTGCC AGTGGCGATA AGTCGTGTCT TACCGGGTTG GACTCAAGAC GATAGTTACC

 2401 GGATAAGGCG CAGCGGTCGG GCTGAACGGG GGGTTCGTGC ACACAGCCCA GCTTGGAGCG

 2461 AACGACCTAC ACCGAACTGA GATACCTACA GCGTGAGCTA TGAGAAAGCG CCACGCTTCC

 2521 CGAAGGGAGA AAGGCGGACA GGTATCCGGT AAGCGGCAGG GTCGGAACAG GAGAGCGCAC

 2581 GAGGGAGCTT CCAGGGGGAA ACGCCTGGTA TCTTTATAGT CCTGTCGGGT TTCGCCACCT

 2641 CTGACTTGAG CGTCGATTTT TGTGATGCTC GTCAGGGGGG CGGAGCCTAT GGAAAAACGC

 2701 CAGCAACGCG GCCTTTTTAC GGTTCCTGGC CTTTTGCTGG CCTTTTGCTC ACATGTTCTT

 2761 TCCTGCGTTA TCCCCTGATT CTGTGGATAA CCGTATTACC GCCTTTGAGT GAGCTGATAC

 2821 CGCTCGCCGC AGCCGAACGA CCGAGCGCAG CGAGTCAGTG AGCGAGGAAG CGGAAGAGCG

 2881 CCTGATGCGG TATTTTCTCC TTACGCATCT GTGCGGTATT TCACACCGCA TATGGTGCAC

 2941 TCTCAGTACA ATCTGCTCTG ATGCCGCATA GTTAAGCCAG TATACACTCC GCTATCGCTA

 3001 CGTGACTGGG TCATGGCTGC GCCCCGACAC CCGCCAACAC CCGCTGACGC GCCCTGACGG

 3061 GCTTGTCTGC TCCCGGCATC CGCTTACAGA CAAGCTGTGA CCGTCTCCGG GAGCTGCATG

 3121 TGTCAGAGGT TTTCACCGTC ATCACCGAAA CGCGCGAGGC AGCAGATCAA TTCGCGCGCG

 3181 AAGGCGAAGC GGCATGCATT TACGTTGACA CCATCGAATG GTGCAAAACC TTTCGCGGTA

 3241 TGGCATGATA GCGCCCGGAA GAGAGTCAAT TCAGGGTGGT GAATGTGAAA CCAGTAACGT

 3301 TATACGATGT CGCAGAGTAT GCCGGTGTCT CTTATCAGAC CGTTTCCCGC GTGGTGAACC

 3361 AGGCCAGCCA CGTTTCTGCG AAAACGCGGG AAAAAGTGGA AGCGGCGATG GCGGAGCTGA

 3421 ATTACATTCC CAACCGCGTG GCACAACAAC TGGCGGGCAA ACAGTCGTTG CTGATTGGCG

 3481 TTGCCACCTC CAGTCTGGCC CTGCACGCGC CGTCGCAAAT TGTCGCGGCG ATTAAATCTC

 3541 GCGCCGATCA ACTGGGTGCC AGCGTGGTGG TGTCGATGGT AGAACGAAGC GGCGTCGAAG

 3601 CCTGTAAAGC GGCGGTGCAC AATCTTCTCG CGCAACGCGT CAGTGGGCTG ATCATTAACT

 3661 ATCCGCTGGA TGACCAGGAT GCCATTGCTG TGGAAGCTGC CTGCACTAAT GTTCCGGCGT

 3721 TATTTCTTGA TGTCTCTGAC CAGACACCCA TCAACAGTAT TATTTTCTCC CATGAAGACG

 3781 GTACGCGACT GGGCGTGGAG CATCTGGTCG CATTGGGTCA CCAGCAAATC GCGCTGTTAG

 3841 CGGGCCCATT AAGTTCTGTC TCGGCGCGTC TGCGTCTGGC TGGCTGGCAT AAATATCTCA

 3901 CTCGCAATCA AATTCAGCCG ATAGCGGAAC GGGAAGGCGA CTGGAGTGCC ATGTCCGGTT

 3961 TTCAACAAAC CATGCAAATG CTGAATGAGG GCATCGTTCC CACTGCGATG CTGGTTGCCA

 4021 ACGATCAGAT GGCGCTGGGC GCAATGCGCG CCATTACCGA GTCCGGGCTG CGCGTTGGTG

 4081 CGGATATCTC GGTAGTGGGA TACGACGATA CCGAAGACAG CTCATGTTAT ATCCCGCCGT

 4141 CAACCACCAT CAAACAGGAT TTTCGCCTGC TGGGGCAAAC CAGCGTGGAC CGCTTGCTGC

 4201 AACTCTCTCA GGGCCAGGCG GTGAAGGGCA ATCAGCTGTT GCCCGTCTCA CTGGTGAAAA

 4261 GAAAAACCAC CCTGGCGCCC AATACGCAAA CCGCCTCTCC CCGCGCGTTG GCCGATTCAT

 4321 TAATGCAGCT GGCACGACAG GTTTCCCGAC TGGAAAGCGG GCAGTGAGCG CAACGCAATT

 4381 AATGTGAGTT AGCGCGAATT GATCTG

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