pBAD-TOPO: Difference between revisions

Latest revision as of 16:56, 30 August 2011

At a Glance

Type	Expression
Origin of Replication	pBR322
Copy #
Markers	Ampicillin
Link to Sequence	Invitrogen
Features	araBAD Promoter RBS Enterokinase site TOPO® Cloning site V5 epitope tag 6xHis tag araC (complementary strand)
Accession Number
edit table

Notes

The pBAD-TOPO vector allows for simple single-step cloning and expression of Taq-amplified PCR products. Upon activation to the linearized state by topoisomerase I, pBAD is ready for quick cloning and subsequent expression in E. coli by the tight regulated araBad promoter. Its C-terminal (6xHis) tag enables purificaton and detection. The V5 epitope tag gives an additional means of detection by Anti-V5 Antibody, and the Enterokinase(EK) cleavage site enables removal of the N-terminal leader peptide.

History

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Sources

Invitrogen

References

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Sequence

AAGAAACCAATTGTCCATATTGCATCAGACATTGCCGTCACTGCGTCTTTTACTGGCTCTTCTCGCTAACCA
AACCGGTAACCCCGCTTATTAAAAGCATTCTGTAACAAAGCGGGACCAAAGCCATGACAAAAACGCGTAACA
AAAGTGTCTATAATCACGGCAGAAAAGTCCACATTGATTATTTGCACGGCGTCACACTTTGCTATGCCATAG
CATTTTTATCCATAAGATTAGCGGATCCTACCTGACGCTTTTTATCGCAACTCTCTACTGTTTCTCCATACC
CGTTTTTTTGGGCTAGAAATAATTTTGTTTAACTTTAAGAAGGAGATATACATACCCATGGGCTCTGGATCC
GGTGATGACGATGACAAGCTCGCCCTTAAGGGCGAGCTTGAAGGTAAGCCTATCCCTAACCCTCTCCTCGGT
CTCGATTCTACGCGTACCGGTCATCATCACCATCACCATTGAGTTTAAACGGTCTCCAGCTTGGCTGTTTTG
GCGGATGAGAGAAGATTTTCAGCCTGATACAGATTAAATCAGAACGCAGAAGCGGTCTGATAAAACAGAATT
TGCCTGGCGGCAGTAGCGCGGTGGTCCCACCTGACCCCATGCCGAACTCAGAAGTGAAACGCCGTAGCGCCG
ATGGTAGTGTGGGGTCTCCCCATGCGAGAGTAGGGAACTGCCAGGCATCAAATAAAACGAAAGGCTCAGTCG
AAAGACTGGGCCTTTCGTTTTATCTGTTGTTTGTCGGTGAACGCTCTCCTGAGTAGGACAAATCCGCCGGGA
GCGGATTTGAACGTTGCGAAGCAACGGCCCGGAGGGTGGCGGGCAGGACGCCCGCCATAAACTGCCAGGCAT
CAAATTAAGCAGAAGGCCATCCTGACGGATGGCCTTTTTGCGTTTCTACAAACTCTTTTTGTTTATTTTTCT
AAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGA
AGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTG
CTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAAC
TGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTA
AAGTTCTGCTATGTGGCGCGGTATTATCCCGTGTTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACT
ATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAG
AATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGAC
CGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGC
TGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAAC
TATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTG
CAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTG
GGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGG
GGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGT
AACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCT
AGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAG
ACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAA
AAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTG
GCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACT
CTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGT
GTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGT
GCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCG
CCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGA
GGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTC
GATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCC
TGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTA
CCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAG
CGGAAGAGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATATGGTGCACTC
TCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGTATACACTCCGCTATCGCTACGTGACTGGGTCAT
GGCTGCGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTA
CAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAG
GCAGCAGATCAATTCGCGCGCGAAGGCGAAGCGGCATGCATAATGTGCCTGTCAAATGGACGAAGCAGGGAT
TCTGCAAACCCTATGCTACTCCGTCAAGCCGTCAATTGTCTGATTCGTTACCAATTATGACAACTTGACGGC
TACATCATTCACTTTTTCTTCACAACCGGCACGGAACTCGCTCGGGCTGGCCCCGGTGCATTTTTTAAATAC
CCGCGAGAAATAGAGTTGATCGTCAAAACCAACATTGCGACCGACGGTGGCGATAGGCATCCGGGTGGTGCT
CAAAAGCAGCTTCGCCTGGCTGATACGTTGGTCCTCGCGCCAGCTTAAGACGCTAATCCCTAACTGCTGGCG
GAAAAGATGTGACAGACGCGACGGCGACAAGCAAACATGCTGTGCGACGCTGGCGATATCAAAATTGCTGTC
TGCCAGGTGATCGCTGATGTACTGACAAGCCTCGCGTACCCGATTATCCATCGGTGGATGGAGCGACTCGTT
AATCGCTTCCATGCGCCGCAGTAACAATTGCTCAAGCAGATTTATCGCCAGCAGCTCCGAATAGCGCCCTTC
CCCTTGCCCGGCGTTAATGATTTGCCCAAACAGGTCGCTGAAATGCGGCTGGTGCGCTTCATCCGGGCGAAA
GAACCCCGTATTGGCAAATATTGACGGCCAGTTAAGCCATTCATGCCAGTAGGCGCGCGGACGAAAGTAAAC
CCACTGGTGATACCATTCGCGAGCCTCCGGATGACGACCGTAGTGATGAATCTCTCCTGGCGGGAACAGCAA
AATATCACCCGGTCGGCAAACAAATTCTCGTCCCTGATTTTTCACCACCCCCTGACCGCGAATGGTGAGATT
GAGAATATAACCTTTCATTCCCAGCGGTCGGTCGATAAAAAAATCGAGATAACCGTTGGCCTCAATCGGCGT
TAAACCCGCCACCAGATGGGCATTAAACGAGTATCCCGGCAGCAGGGGATCATTTTGCGCTTCAGCCATACT
TTTCATACTCCCGCCATTCAGAG

@@ Line 1: / Line 1: @@
 ==At a Glance ==
-[[Image:#PBADTOPO.jpg|thumb|400px|right|pBAD-TOPO, Image provided by [http://www.invitrogen.com/content/sfs/vectors/pbadtopo_map.pdf Invitrogen]]]
+[[File:PBAD-TOPO.jpg|thumb|550px|right|pBAD-TOPO, Image made by [[PlasMapper]]]]
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-{|{{Prettytable}} width='30%' align=left
+{| {{Prettytable}} width='30%' align=left  id="J47051bd697d3d"  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/pbadtopo_seq.txt, Invitrogen]
+[http://tools.invitrogen.com/content/sfs/vectors/pbadtopo_seq.txt Invitrogen]
 |-
-!{{table heading style}} |Comments
+!align=left {{table heading style}} |Features
-|
+||
+araBAD Promoter <br />
+RBS <br />
+Enterokinase site <br />
+TOPO® Cloning site <br />
+V5 epitope tag <br />
+xHis tag <br />
+araC (complementary strand) <br />
+|-
+!align=left {{table heading style}} |Accession Number
+||
-|-
-|colspan='2'|[{{SERVER}}/colipedia/index.php?title=Special:TableEdit&id=63ce1649aab8b3180235b0af7618f667.0.J47051bd697d3d&page=0&pagename={{FULLPAGENAMEE}} edit table]
+|- class="tableEdit_footer"
+|<span class="tableEdit_editLink plainlinks">[{{SERVER}}{{SCRIPTPATH}}?title=Special:TableEdit&id=63ce1649aab8b3180235b0af7618f667.0.J47051bd697d3d&page=112770&pagename={{FULLPAGENAMEE}}&type=1&template=PlasmidTable edit table]</span> ||
 |}
 <!--box uid=63ce1649aab8b3180235b0af7618f667.0.J47051bd697d3d--></protect>
 <BR>
 <Br clear='all'>
+==Notes==
+The pBAD-TOPO vector allows for simple single-step cloning and expression of Taq-amplified PCR products. Upon activation to the linearized state by topoisomerase I, pBAD is ready for quick cloning and subsequent expression in E. coli by the tight regulated araBad promoter. Its C-terminal (6xHis) tag enables purificaton and detection. The V5 epitope tag gives an additional means of detection by Anti-V5 Antibody, and the Enterokinase(EK) cleavage site enables removal of the N-terminal leader peptide.
 ==History {{HelpIcon|Help:Plasmid}}==
@@ Line 44: / Line 59: @@
 ==Sources ==
-[http://www.invitrogen.com/content/sfs/vectors/pbadtopo_map.pdf, Invitrogen]
+[http://www.invitrogen.com/site/us/en/home.html  Invitrogen]
 ==References==
+{{BacktoTop}}
 {{RefHelp}}
-<references/>
+<references />
 ==Sequence==
@@ Line 114: / Line 130: @@
-[[Category:Plasmids]][[Category:Expression Plasmids]][[Category:Invitrogen Plasmids]][[Category:Ampicillin Marker Plasmids]]
+[[Category:Plasmids]][[Category:Expression Plasmids]][[Category:Invitrogen Plasmids]][[Category:Ampicillin Marker Plasmids]][[Category:araBAD Promoter]][[Category:Enterokinase site]][[Category:LacZ plasmids]][[Category:V5 Tag]][[Category: 6X His Tag]][[Category:araC]]
 __NOTOC__