pTrcHis2 B: Difference between revisions
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!{{table heading style}} |Type | !align=left {{table heading style}} |Type | ||
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Expression | Expression | ||
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!{{table heading style}} |Origin of Replication | !align=left {{table heading style}} |Origin of Replication | ||
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pBR322 | pBR322 | ||
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!{{table heading style}} |Copy # | !align=left {{table heading style}} |Copy # | ||
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!{{table heading style}} |Markers | !align=left {{table heading style}} |Markers | ||
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Ampicillin | Ampicillin | ||
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!{{table heading style}} |Link to Sequence | !align=left {{table heading style}} |Link to Sequence | ||
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[http://www.invitrogen.com/content/sfs/vectors/ptrchis2b_seq.txt Invitrogen] | [http://www.invitrogen.com/content/sfs/vectors/ptrchis2b_seq.txt Invitrogen] | ||
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!{{table heading style}} |Features | !align=left {{table heading style}} |Features | ||
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trc promoter | trc promoter | ||
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''Xba''I | ''Xba''I | ||
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!{{table heading style}} |Accession Number | !align=left {{table heading style}} |Accession Number | ||
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Revision as of 09:44, 20 February 2009
At a Glance
[[Image:#pTrcHis2 A-C.JPG|thumb|right|200px|pTrcHis2 A-C, image provided by Invitrogen]]
| Type |
Expression |
|---|---|
| Origin of Replication |
pBR322 |
| Copy # | |
| Markers |
Ampicillin |
| Link to Sequence | |
| Features |
trc promoter lac operator rrnB antitermination region Bacteriophage gene 10 translational enhancer Minicistron and reinitiation ribosome binding site Initiation ATG Multiple cloning site C-terminal myc epitope tag C-terminal polyhistidine region Ampicillin resistance gene XbaI |
| Accession Number |
|
| edit table |
Notes
PTrcHis2 B is a pBR322-derived expression vector. High levels of expression are possible using the trc promoter and the rrnB anti-termination region. The trc promoter contains the -35 region of the trp promoter together with the -10 region of the lac promoter. To regulate expression, the gene encoding Lac repressor (lacIq) is provided in the pTrcHis2 vectors, allowing regulation of the trc promoter. 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.
PtrcHis2 B differs from the A and C strains because it is the only one of the three that contains the Xba I gene.
History
You can help EcoliWiki by writing about the history of this plasmid.
Sources
References
- ↑ Li, SC et al. (1984) Antitermination of E. coli rRNA transcription is caused by a control region segment containing lambda nut-like sequences. Cell 38 851-60 PubMed
- ↑ Schoner, BE et al. (1986) Translation of a synthetic two-cistron mRNA in Escherichia coli. Proc. Natl. Acad. Sci. U.S.A. 83 8506-10 PubMed
- ↑ Brosius, J et al. (1985) Spacing of the -10 and -35 regions in the tac promoter. Effect on its in vivo activity. J. Biol. Chem. 260 3539-41 PubMed
- ↑ Amann, E et al. (1983) Vectors bearing a hybrid trp-lac promoter useful for regulated expression of cloned genes in Escherichia coli. Gene 25 167-78 PubMed
Sequence
GTTTGACAGCTTATCATCGACTGCACGGTGCACCAATGCTTCTGGCGTCAGGCA GCCATCGGAAGCTGTGGTATGGCTGTGCAGGTCGTAAATCACTGCATAATTCG TGTCGCTCAAGGCGCACTCCCGTTCTGGATAATGTTTTTTGCGCCGACATCATA ACGGTTCTGGCAAATATTCTGAAATGAGCTGTTGACAATTAATCATCCGGCTCG TATAATGTGTGGAATTGTGAGCGGATAACAATTTCACACAGGAAACAGCGCCG CTGAGAAAAAGCGAAGCGGCACTGCTCTTTAACAATTTATCAGACAATCTGTGT GGGCACTCGACCGGAATTATCGATTAACTTTATTATTAAAAATTAAAGAGGTAT ATATTAATGTATCGATTAAATAAGGAGGAATAAACCATGGATCCGAGCTCGAGA TCTGCAGCTGGTACCATATGGGAATTCGAAGCTTTCTAGAACAAAAACTCATCT CAGAAGAGGATCTGAATAGCGCCGTCGACCATCATCATCATCATCATTGAGTTT AAACGGTCTCCAGCTTGGCTGTTTTGGCGGATGAGAGAAGATTTTCAGCCTGAT ACAGATTAAATCAGAACGCAGAAGCGGTCTGATAAAACAGAATTTGCCTGGCG GCAGTAGCGCGGTGGTCCCACCTGACCCCATGCCGAACTCAGAAGTGAAACGC CGTAGCGCCGATGGTAGTGTGGGGTCTCCCCATGCGAGAGTAGGGAACTGCC AGGCATCAAATAAAACGAAAGGCTCAGTCGAAAGACTGGGCCTTTCGTTTTATC TGTTGTTTGTCGGTGAACGCTCTCCTGAGTAGGACAAATCCGCCGGGAGCGGA TTTGAACGTTGCGAAGCAACGGCCCGGAGGGTGGCGGGCAGGACGCCCGCCA TAAACTGCCAGGCATCAAATTAAGCAGAAGGCCATCCTGACGGATGGCCTTTTT GCGTTTCTACAAACTCTTTTTGTTTATTTTTCTAAATACATTCAAATATGTATCCG CTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGT ATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCC TTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATC AGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATC CTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTT CTGCTATGTGGCGCGGTATTATCCCGTGTTGACGCCGGGCAAGAGCAACTCGG TCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGA AAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAA CCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCG AAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGAT CGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCA CGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTA CTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTT GCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAA TCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAG ATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACT ATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCA TTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTT CATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCA AAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGA TCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAAC AAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAA CTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCC TTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTA CATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGT CGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGG TCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCT ACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCC GAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGA GAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGT CGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGG GGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCC TTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGG ATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACG ACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCTGATGCGG TATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATATGGTGCACTCTCA GTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGTATACACTCCGCTATCGCT ACGTGACTGGGTCATGGCTGCGCCCCGACACCCGCCAACACCCGCTGACGCG CCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGT CTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGA GGCAGCAGATCAATTCGCGCGCGAAGGCGAAGCGGCATGCATTTACGTTGACA CCATCGAATGGTGCAAAACCTTTCGCGGTATGGCATGATAGCGCCCGGAAGAG AGTCAATTCAGGGTGGTGAATGTGAAACCAGTAACGTTATACGATGTCGCAGA GTATGCCGGTGTCTCTTATCAGACCGTTTCCCGCGTGGTGAACCAGGCCAGCC ACGTTTCTGCGAAAACGCGGGAAAAAGTGGAAGCGGCGATGGCGGAGCTGAA TTACATTCCCAACCGCGTGGCACAACAACTGGCGGGCAAACAGTCGTTGCTGA TTGGCGTTGCCACCTCCAGTCTGGCCCTGCACGCGCCGTCGCAAATTGTCGCG GCGATTAAATCTCGCGCCGATCAACTGGGTGCCAGCGTGGTGGTGTCGATGGT AGAACGAAGCGGCGTCGAAGCCTGTAAAGCGGCGGTGCACAATCTTCTCGCG CAACGCGTCAGTGGGCTGATCATTAACTATCCGCTGGATGACCAGGATGCCAT TGCTGTGGAAGCTGCCTGCACTAATGTTCCGGCGTTATTTCTTGATGTCTCTGA CCAGACACCCATCAACAGTATTATTTTCTCCCATGAAGACGGTACGCGACTGGG CGTGGAGCATCTGGTCGCATTGGGTCACCAGCAAATCGCGCTGTTAGCGGGCC CATTAAGTTCTGTCTCGGCGCGTCTGCGTCTGGCTGGCTGGCATAAATATCTCA CTCGCAATCAAATTCAGCCGATAGCGGAACGGGAAGGCGACTGGAGTGCCAT GTCCGGTTTTCAACAAACCATGCAAATGCTGAATGAGGGCATCGTTCCCACTGC GATGCTGGTTGCCAACGATCAGATGGCGCTGGGCGCAATGCGCGCCATTACCG AGTCCGGGCTGCGCGTTGGTGCGGATATCTCGGTAGTGGGATACGACGATACC GAAGACAGCTCATGTTATATCCCGCCGTCAACCACCATCAAACAGGATTTTCGC CTGCTGGGGCAAACCAGCGTGGACCGCTTGCTGCAACTCTCTCAGGGCCAGGC GGTGAAGGGCAATCAGCTGTTGCCCGTCTCACTGGTGAAAAGAAAAACCACCC TGGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATG CAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCA ATTAATGTGAGTTAGCGCGAATTGATCTG