Difference between revisions of "Part:BBa K404205"
 
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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K404205 short</partinfo>
 
<partinfo>BBa_K404205 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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{| 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_K404205 ViralBrick-587-BAP]
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|-
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|'''BioBrick Nr.'''
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|[https://parts.igem.org/Part:BBa_K404165 BBa_K404205]
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|-
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|'''RFC standard'''
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|[https://parts.igem.org/Help:Assembly_standard_25 RFC 25]
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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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|
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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/><br/><br/><br/><br/><br/><br/><br/>
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<br><b>The Biotinylation Acceptor peptide motif, ready for insertion into the 587 loop</b>
  
 
[[Image:Freiburg10_ViralBrick-logo-587-BAP.png|thumb|center|480px]]<br>
 
[[Image:Freiburg10_ViralBrick-logo-587-BAP.png|thumb|center|480px]]<br>
  
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<!-- Add more about the biology of this part here
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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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===Restriction sites===
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<table border="1" style="float:right; width:480px; height:auto;>
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<tr>
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  <th><b>Restriction site</b></th>
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  <th><b>Enzyme</b></th>
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  <th><b>Recognition sequence</b></th>
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  <th><b>Activity</b></th>
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  <th><b>Temp.</b></th>
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</tr>
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<tr>
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  <td>upstream 587</td>
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  <td>[http://www.neb.com/nebecomm/products/productR3136.asp BamHI-HF]</td>
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  <td>[[image:Freiburg10_recognition site BamHI-HF.Gif]]</td>
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  <td>100;50;10;100</td>
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  <td>37°C</td>
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</tr>
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<tr>
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  <td>downstream 587</td>
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  <td>[http://www.neb.com/nebecomm/products/productR3151.asp PvuII-HF]</td>
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  <td>[[image:Freiburg10_recognition site PvuII-HF.Gif]]</td>
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  <td>0;25;0;100</td>
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  <td>37°C
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</tr>
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<tr>
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</table>
  
 
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Latest revision as of 02:23, 28 October 2010

ViralBrick-587-BAP

ViralBrick-587-BAP
BioBrick Nr. BBa_K404205
RFC standard RFC 25
Requirement pSB1C3
Source
Submitted by [http://2010.igem.org/Team:Freiburg_Bioware FreiGEM 2010]















The Biotinylation Acceptor peptide motif, ready for insertion into the 587 loop

Freiburg10 ViralBrick-logo-587-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).

Restriction sites

Restriction site Enzyme Recognition sequence Activity Temp.
upstream 587 [http://www.neb.com/nebecomm/products/productR3136.asp BamHI-HF] Freiburg10 recognition site BamHI-HF.Gif 100;50;10;100 37°C
downstream 587 [http://www.neb.com/nebecomm/products/productR3151.asp PvuII-HF] Freiburg10 recognition site PvuII-HF.Gif 0;25;0;100 37°C

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 10
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]