pTrcHis2 B: Difference between revisions

Latest revision as of 21:10, 22 August 2011

At a Glance

Type	Expression
Origin of Replication	pBR322
Copy #
Markers	Ampicillin
Link to Sequence	Invitrogen
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

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Sources

Invitrogen

References

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^[1] ^[2] ^[3] ^[4]

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

↑ 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

[PMID:6091902-1] 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

[PMID:3534891-2] 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

[PMID:2579077-3] 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

[PMID:6363212-4] 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

[1]

[2]

[3]

[4]

@@ Line 1: / Line 1: @@
 ==At a Glance==
-[[Image:#pTrcHis2 A-C.JPG|thumb|right|200px|pTrcHis2 A-C, image provided by [http://www.invitrogen.com/content/sfs/vectors/ptrchis2_map.pdf Invitrogen]]]
+[[File:PTrcHis2 B.jpg|thumb|450px|pTrcHis2 B, Image made with [[PlasMapper]]]]
 <protect><!--box uid=63ce1649aab8b3180235b0af7618f667.0.M470bc5c9d909c-->
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 ******************************************************************************************
 *
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-{|{{Prettytable}} width='30%' align=left
+{| {{Prettytable}} width='30%' align=left  id="M470bc5c9d909c"  class=" tableEdit PlasmidTable"
 |-
-!{{table heading style}} |Type
+!align=left {{table heading style}} |Type
-|
+||
 Expression
 |-
-!{{table heading style}} |Origin of Replication
+!align=left {{table heading style}} |Origin of Replication
-|
+||
 pBR322
 |-
-!{{table heading style}} |Copy #
+!align=left {{table heading style}} |Copy #
-|
+||
 |-
-!{{table heading style}} |Markers
+!align=left {{table heading style}} |Markers
-|
+||
 Ampicillin
 |-
-!{{table heading style}} |Link to Sequence
+!align=left {{table heading style}} |Link to Sequence
-|
+||
 [http://www.invitrogen.com/content/sfs/vectors/ptrchis2b_seq.txt Invitrogen]
 |-
-!{{table heading style}} |Comments
+!align=left {{table heading style}} |Features
-|
+||
+trc promoter<Br />
+lac operator<Br />
+rrnB antitermination region<Br />
+Bacteriophage gene 10 translational enhancer<Br />
+Minicistron and reinitiation ribosome binding site<Br />
+Initiation ATG<Br />
+Multiple cloning site<Br />
+C-terminal myc epitope tag<Br />
+C-terminal polyhistidine region<Br />
+Ampicillin resistance gene<Br />
+''Xba''I<Br />
+|-
+!align=left {{table heading style}} |Accession Number
+||
-|-
+|- class="tableEdit_footer"
-|colspan='2'|[{{SERVER}}/colipedia/index.php?title=Special:TableEdit&id=63ce1649aab8b3180235b0af7618f667.0.M470bc5c9d909c&page=0&pagename={{FULLPAGENAMEE}} edit table]
+|<span class="tableEdit_editLink plainlinks">[{{SERVER}}{{SCRIPTPATH}}?title=Special:TableEdit&id=63ce1649aab8b3180235b0af7618f667.0.M470bc5c9d909c&page=112918&pagename={{FULLPAGENAMEE}}&type=1&template=PlasmidTable edit table]</span> ||
 |}
 <!--box uid=63ce1649aab8b3180235b0af7618f667.0.M470bc5c9d909c--></protect>
 <BR>
 <Br clear='all'>
+==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 {{HelpIcon|Help:Plasmid}}==
@@ Line 47: / Line 68: @@
 ==References==
+{{BacktoTop}}
 {{RefHelp}}
-<references/>
+<references />
+<ref name='PMID:6091902'/>
+<ref name='PMID:3534891'/>
+<ref name='PMID:2579077'/>
+<ref name='PMID:6363212'/>
 ==Sequence==
 <pre>
@@ Line 142: / Line 167: @@
-[[Category:Plasmids]][[Category:Expression Plasmids]][[Category:Invitrogen Plasmids]]
+[[Category:Plasmids]][[Category:Expression Plasmids]][[Category:Invitrogen Plasmids]][[Category:Ampicillin Marker Plasmids]]
 __NOTOC__