pTrcHis2 B: Difference between revisions

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==At a Glance==
==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]]]]
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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
Expression
|-
|-
!{{table heading style}} |Origin of Replication
!align=left {{table heading style}} |Origin of Replication
|
||
pBR322
pBR322
|-
|-
!{{table heading style}} |Copy #
!align=left {{table heading style}} |Copy #
|
||


|-
|-
!{{table heading style}} |Markers
!align=left {{table heading style}} |Markers
|
||
Ampicillin
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]
[http://www.invitrogen.com/content/sfs/vectors/ptrchis2b_seq.txt Invitrogen]
|-
|-
!{{table heading style}} |Features
!align=left {{table heading style}} |Features
|
||
trc promoter
trc promoter<Br />
 
lac operator<Br />
lac operator
rrnB antitermination region<Br />
 
Bacteriophage gene 10 translational enhancer<Br />
rrnB antitermination region
Minicistron and reinitiation ribosome binding site<Br />
 
Initiation ATG<Br />
Bacteriophage gene 10 translational enhancer
Multiple cloning site<Br />
 
C-terminal myc epitope tag<Br />
Minicistron and reinitiation ribosome binding site
C-terminal polyhistidine region<Br />
 
Ampicillin resistance gene<Br />
Initiation ATG
''Xba''I<Br />
 
|-
Multiple cloning site
!align=left {{table heading style}} |Accession Number
 
||
C-terminal myc epitope tag
 
C-terminal polyhistidine region
 
Ampicillin resistance gene


''Xba''I
|-
!{{table heading style}} |Accession Number
|


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==Notes==
==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 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.
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==References==
==References==
 
{{BacktoTop}}
{{RefHelp}}
<references />
<ref name='PMID:6091902'/>
<ref name='PMID:6091902'/>
<ref name='PMID:3534891'/>
<ref name='PMID:3534891'/>
<ref name='PMID:2579077'/>
<ref name='PMID:2579077'/>
<ref name='PMID:6363212'/>
<ref name='PMID:6363212'/>
<references/>
==Sequence==
==Sequence==
<pre>
<pre>

Latest revision as of 21:10, 22 August 2011

At a Glance

pTrcHis2 B, Image made with PlasMapper
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

You can help EcoliWiki by writing about the history of this plasmid.

Sources

Invitrogen

References

[back to top]


See Help:References for how to manage references in EcoliWiki.

[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
  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
  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
  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
  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
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