pBAD/gIII C: Difference between revisions

Latest revision as of 16:33, 23 August 2011

At a Glance

Type	Expression
Origin of Replication	pBR322
Copy #
Markers	Ampicillin
Link to Sequence	Invitrogen
Features	araBAD promoter Optimized ribosome binding site Initiation ATG MCS (multiple cloning site) C-terminal polyhistidine region araC gene
Accession Number
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Notes

The pBAD/gIII Kit provides all of the reagents to express recombinant proteins under the tight regulation of the araBAD promoter. To simplify purification, the vector pBAD/gIII encodes a leader peptide which directs the recombinant protein into theperiplasmic space. The vector provides the araBAD promoter for tightly regulated expression, translation initiation signals for optimized E. coli expression, leader peptide from the bacteriophage fd gene III protein (gIII) for secreted expression, C-terminal polyhistidine (6xHis) tag for purification with nickel-chelating resin, and C-terminal c-myc epitope for detection and analysis with an Anti-myc Antibody.

A set of three vectors is provided (A, B, and C). Each has the C-terminal tag in a different reading frame relative to the multiple cloning site to simplify in-frame cloning of your gene.

pBAD/gIII Version C contains Not I only.

History

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Sources

Invitrogen

References

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

Sequence

AAGAAACCAATTGTCCATATTGCATCAGACATTGCCGTCACTGCGTCTTTTACTGGCTCTTCTCGCTAACCA
AACCGGTAACCCCGCTTATTAAAAGCATTCTGTAACAAAGCGGGACCAAAGCCATGACAAAAACGCGTAACA
AAAGTGTCTATAATCACGGCAGAAAAGTCCACATTGATTATTTGCACGGCGTCACACTTTGCTATGCCATAG
CATTTTTATCCATAAGATTAGCGGATCCTACCTGACGCTTTTTATCGCAACTCTCTACTGTTTCTCCATACC
CGTTTTTTGGGCTAACAGGAGGAATTAACCATGAAAAAACTGCTGTTCGCGATTCCGCTGGTGGTGCCGTTC
TATAGCCATAGCACCATGGCGGCCGCTCGAGATCTGCAGCTGGTACCATATGGGAATTCGAAGCTTTCTAGA
ACAAAAACTCATCTCAGAAGAGGATCTGAATAGCGCCGTCGACCATCATCATCATCATCATTGAGTTTAAAC
GGTCTCCAGCTTGGCTGTTTTGGCGGATGAGAGAAGATTTTCAGCCTGATACAGATTAAATCAGAACGCAGA
AGCGGTCTGATAAAACAGAATTTGCCTGGCGGCAGTAGCGCGGTGGTCCCACCTGACCCCATGCCGAACTCA
GAAGTGAAACGCCGTAGCGCCGATGGTAGTGTGGGGTCTCCCCATGCGAGAGTAGGGAACTGCCAGGCATCA
AATAAAACGAAAGGCTCAGTCGAAAGACTGGGCCTTTCGTTTTATCTGTTGTTTGTCGGTGAACGCTCTCCT
GAGTAGGACAAATCCGCCGGGAGCGGATTTGAACGTTGCGAAGCAACGGCCCGGAGGGTGGCGGGCAGGACG
CCCGCCATAAACTGCCAGGCATCAAATTAAGCAGAAGGCCATCCTGACGGATGGCCTTTTTGCGTTTCTACA
AACTCTTTTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATG
CTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCG
GCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGT
GCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGT
TTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTGTTGACGCCGGGCAAGAG
CAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTT
ACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTA
CTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGC
CTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCA
ATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGAC
TGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGAT
AAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGT
ATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGT
GCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTT
CATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAG
TTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGC
GTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCA
ACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAG
TTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCT
GCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGG
TCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTA
CAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGG
GTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTT
CGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGC
AACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCT
GATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGC
AGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATT
TCACACCGCATATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGTATACACTCCGC
TATCGCTACGTGACTGGGTCATGGCTGCGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTT
GTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCAC
CGTCATCACCGAAACGCGCGAGGCAGCAGATCAATTCGCGCGCGAAGGCGAAGCGGCATGCATAATGTGCCT
GTCAAATGGACGAAGCAGGGATTCTGCAAACCCTATGCTACTCCGTCAAGCCGTCAATTGTCTGATTCGTTA
CCAATTATGACAACTTGACGGCTACATCATTCACTTTTTCTTCACAACCGGCACGGAACTCGCTCGGGCTGG
CCCCGGTGCATTTTTTAAATACCCGCGAGAAATAGAGTTGATCGTCAAAACCAACATTGCGACCGACGGTGG
CGATAGGCATCCGGGTGGTGCTCAAAAGCAGCTTCGCCTGGCTGATACGTTGGTCCTCGCGCCAGCTTAAGA
CGCTAATCCCTAACTGCTGGCGGAAAAGATGTGACAGACGCGACGGCGACAAGCAAACATGCTGTGCGACGC
TGGCGATATCAAAATTGCTGTCTGCCAGGTGATCGCTGATGTACTGACAAGCCTCGCGTACCCGATTATCCA
TCGGTGGATGGAGCGACTCGTTAATCGCTTCCATGCGCCGCAGTAACAATTGCTCAAGCAGATTTATCGCCA
GCAGCTCCGAATAGCGCCCTTCCCCTTGCCCGGCGTTAATGATTTGCCCAAACAGGTCGCTGAAATGCGGCT
GGTGCGCTTCATCCGGGCGAAAGAACCCCGTATTGGCAAATATTGACGGCCAGTTAAGCCATTCATGCCAGT
AGGCGCGCGGACGAAAGTAAACCCACTGGTGATACCATTCGCGAGCCTCCGGATGACGACCGTAGTGATGAA
TCTCTCCTGGCGGGAACAGCAAAATATCACCCGGTCGGCAAACAAATTCTCGTCCCTGATTTTTCACCACCC
CCTGACCGCGAATGGTGAGATTGAGAATATAACCTTTCATTCCCAGCGGTCGGTCGATAAAAAAATCGAGAT
AACCGTTGGCCTCAATCGGCGTTAAACCCGCCACCAGATGGGCATTAAACGAGTATCCCGGCAGCAGGGGAT
CATTTTGCGCTTCAGCCATACTTTTCATACTCCCGCCATTCAGAG

↑ San Millan, JL et al. (1989) Use of phoA fusions to study the topology of the Escherichia coli inner membrane protein leader peptidase. J. Bacteriol. 171 5536-41 PubMed
↑ Boeke, JD & Model, P (1982) A prokaryotic membrane anchor sequence: carboxyl terminus of bacteriophage f1 gene III protein retains it in the membrane. Proc. Natl. Acad. Sci. U.S.A. 79 5200-4 PubMed
↑ Lee, N et al. (1987) Arabinose-induced binding of AraC protein to araI2 activates the araBAD operon promoter. Proc. Natl. Acad. Sci. U.S.A. 84 8814-8 PubMed

[PMID:2551889-1] San Millan, JL et al. (1989) Use of phoA fusions to study the topology of the Escherichia coli inner membrane protein leader peptidase. J. Bacteriol. 171 5536-41 PubMed

[PMID:6291030-2] Boeke, JD & Model, P (1982) A prokaryotic membrane anchor sequence: carboxyl terminus of bacteriophage f1 gene III protein retains it in the membrane. Proc. Natl. Acad. Sci. U.S.A. 79 5200-4 PubMed

[PMID:2962192-3] Lee, N et al. (1987) Arabinose-induced binding of AraC protein to araI2 activates the araBAD operon promoter. Proc. Natl. Acad. Sci. U.S.A. 84 8814-8 PubMed

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[2]

[3]

@@ Line 1: / Line 1: @@
-==At a Glance {{HelpIcon|Help:Plasmid}}==
+==At a Glance==
+[[File:pBAD-gIII C.jpg|thumb|450px|pBAD/gIII C, Image made with [[PlasMapper]]]]
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+{| {{Prettytable}} width='30%' align=left  id="N4713e683eb6b8"  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
-|
+||
 [https://www.invitrogen.com/content/sfs/vectors/pbadgiiic_seq.txt Invitrogen]
 |-
-!{{table heading style}} |Comments
+!align=left {{table heading style}} |Features
-|
+||
+araBAD promoter<Br />
+Optimized ribosome binding site<Br />
+Initiation ATG<Br />
+MCS (multiple cloning site)<Br />
+C-terminal polyhistidine region<Br />
+araC gene<Br />
+|-
+!align=left {{table heading style}} |Accession Number
+||
-|-
-|colspan='2'|[{{SERVER}}/colipedia/index.php?title=Special:TableEdit&id=63ce1649aab8b3180235b0af7618f667.0.N4713e683eb6b8&page=0&pagename={{FULLPAGENAMEE}} edit table]
+|- class="tableEdit_footer"
+|<span class="tableEdit_editLink plainlinks">[{{SERVER}}{{SCRIPTPATH}}?title=Special:TableEdit&id=63ce1649aab8b3180235b0af7618f667.0.N4713e683eb6b8&page=113028&pagename={{FULLPAGENAMEE}}&type=1&template=PlasmidTable edit table]</span> ||
 |}
 <!--box uid=63ce1649aab8b3180235b0af7618f667.0.N4713e683eb6b8--></protect>
 <BR>
 <Br clear='all'>
+==Notes==
+The pBAD/gIII Kit provides all of the reagents to express recombinant proteins under the tight regulation of the araBAD promoter. To simplify purification, the vector pBAD/gIII encodes a leader peptide which directs the recombinant protein into theperiplasmic space. The vector provides the araBAD promoter for tightly regulated expression, translation initiation signals for optimized E. coli expression, leader peptide from the bacteriophage fd gene III protein (gIII) for secreted expression, C-terminal polyhistidine (6xHis) tag for purification with nickel-chelating resin, and C-terminal c-myc epitope for detection and analysis with an Anti-myc Antibody.
+A set of three vectors is provided (A, B, and C). Each has the C-terminal tag in a different reading frame relative to the multiple cloning site to simplify in-frame cloning of your gene.
+pBAD/gIII Version C contains ''Not'' I only.
 ==History {{HelpIcon|Help:Plasmid}}==
 {{PlasmidHistory}}
@@ Line 45: / Line 65: @@
 ==References==
+{{BacktoTop}}
 {{RefHelp}}
-<references/>
+<references />
+<ref name='PMID:2551889'/>
+<ref name='PMID:6291030'/>
+<ref name='PMID:2962192'/>
 ==Sequence==
   AAGAAACCAATTGTCCATATTGCATCAGACATTGCCGTCACTGCGTCTTTTACTGGCTCTTCTCGCTAACCA
@@ Line 112: / Line 135: @@
-[[Category:Plasmids]]
+[[Category:Plasmids]][[Category:Expression Plasmids]][[Category:Invitrogen Plasmids]][[Category:Ampicillin Marker Plasmids]][[Category:araBAD Promoter]][[Category:Optimized RBS]][[Category:6X His Tag]][[Category:MCS]]
 __NOTOC__