Difference between revisions of "Part:BBa K404251"
 
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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K404251 short</partinfo>
 
<partinfo>BBa_K404251 short</partinfo>
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<br><b>The Biotinylation Acceptor peptide motif, ready for insertion into the 453 loop</b>
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<br><b>The Biotinylation Acceptor peptide motif, including a knockout of the natural HSPG tropism, inserted into the 587 loop of [[Part:BBa_K404007|pCMV_[AAV2]-VP123]]</b>
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{| style="color:black" cellpadding="6" cellspacing="1" border="2" align="left"
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! colspan="2" style="background:#66bbff;"|[https://parts.igem.org/Part:BBa_K404251 [AAV2]-left-ITR_pCMV]
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|-
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|'''BioBrick Nr.'''
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|[https://parts.igem.org/Part:BBa_K404251 BBa_K404251]
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|-
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|'''RFC standard'''
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|[https://parts.igem.org/Help:Assembly_standard_10 RFC 10]
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|-
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|'''Requirement'''
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|pSB1C3<br>
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|-
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|'''Source'''
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|pAAV_MCS provided by Stratagene
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|-
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|'''Submitted by'''
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|[http://2010.igem.org/Team:Freiburg_Bioware FreiGEM 2010]
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|}
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<br /><br /><br /><br /><br /><br />
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[[Image:Freiburg10_pCMV_VP123 587KO-BAP.png|thumb|center|480px]]<br>
 
[[Image:Freiburg10_pCMV_VP123 587KO-BAP.png|thumb|center|480px]]<br>
  
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<!-- Add more about the biology of this part here
 
 
===Usage and Biology===
 
===Usage and Biology===
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<html>
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<div style="width:965px; height:400px; ">
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<div style="float:left; width:460px; height:auto;  margin: 0px 5px 0px 5px; text-align:justify;">
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The Biotinylation Acceptor Peptide (BAP) is a 15 amino acid long peptide identified by Schatz J., 1993 in an library screening approach. This peptide with the sequence 5' - GLNDIFEAQKIEWHE - 3' contains a central lysine that is specifically biotinylated by the prokaryotic enzyme biotin holenzyme synthetase, encoded in the BirA gene of E. coli. Specific biotinylation of this peptide sequence can be performed in vivo by contransfecting a plasmid with the BirA gene as described for the AAV in Arnold et al.; 2006 or by an in vitro coupling approach using the purified Escherichia coli enzyme biotin ligase (BirA).
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</div>
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<div style="float:right; width:480px; height:auto; "><img src="https://static.igem.org/mediawiki/parts/a/a9/Freiburg10_ViralBrick_motif_BAP.png" width="460"
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height="auto"/></div></div><br>
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</html>
  
 
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Latest revision as of 16:20, 13 January 2011

pCMV_[AAV2]-VP123ex (ViralBrick-587KO-BAP)
The Biotinylation Acceptor peptide motif, including a knockout of the natural HSPG tropism, inserted into the 587 loop of pCMV_[AAV2]-VP123

[AAV2-left-ITR_pCMV]
BioBrick Nr. BBa_K404251
RFC standard RFC 10
Requirement pSB1C3
Source pAAV_MCS provided by Stratagene
Submitted by [http://2010.igem.org/Team:Freiburg_Bioware FreiGEM 2010]








Freiburg10 pCMV VP123 587KO-BAP.png

Usage and Biology

The Biotinylation Acceptor Peptide (BAP) is a 15 amino acid long peptide identified by Schatz J., 1993 in an library screening approach. This peptide with the sequence 5' - GLNDIFEAQKIEWHE - 3' contains a central lysine that is specifically biotinylated by the prokaryotic enzyme biotin holenzyme synthetase, encoded in the BirA gene of E. coli. Specific biotinylation of this peptide sequence can be performed in vivo by contransfecting a plasmid with the BirA gene as described for the AAV in Arnold et al.; 2006 or by an in vitro coupling approach using the purified Escherichia coli enzyme biotin ligase (BirA).

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 2396
    Illegal XhoI site found at 698
    Illegal XhoI site found at 884
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 665
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 2982
    Illegal SapI site found at 1833