Difference between revisions of "Part:BBa K404205"
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__NOTOC__ | __NOTOC__ | ||
<partinfo>BBa_K404205 short</partinfo> | <partinfo>BBa_K404205 short</partinfo> | ||
| + | {| style="color:black" cellpadding="6" cellspacing="1" border="2" align="left" | ||
| + | ! colspan="2" style="background:#66bbff;"|[https://parts.igem.org/Part:BBa_K404205 ViralBrick-587-BAP] | ||
| + | |- | ||
| + | |'''BioBrick Nr.''' | ||
| + | |[https://parts.igem.org/Part:BBa_K404165 BBa_K404205] | ||
| + | |- | ||
| + | |'''RFC standard''' | ||
| + | |[https://parts.igem.org/Help:Assembly_standard_25 RFC 25] | ||
| + | |- | ||
| + | |'''Requirement''' | ||
| + | |pSB1C3<br> | ||
| + | |- | ||
| + | |'''Source''' | ||
| + | | | ||
| + | |- | ||
| + | |'''Submitted by''' | ||
| + | |[http://2010.igem.org/Team:Freiburg_Bioware FreiGEM 2010] | ||
| + | |} | ||
| + | <br/><br/><br/><br/><br/><br/><br/><br/><br/><br/><br/><br/><br/> | ||
<br><b>The Biotinylation Acceptor peptide motif, ready for insertion into the 587 loop</b> | <br><b>The Biotinylation Acceptor peptide motif, ready for insertion into the 587 loop</b> | ||
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<!-- Add more about the biology of this part here --> | <!-- Add more about the biology of this part here --> | ||
===Usage and Biology=== | ===Usage and Biology=== | ||
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| + | <div style="width:965px; height:400px; "> | ||
| + | <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). | 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). | ||
| + | </div> | ||
<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" | <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" | ||
| − | height="auto"/></div> | + | height="auto"/></div></div><br> |
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| + | </html> | ||
===Restriction sites=== | ===Restriction sites=== | ||
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
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] |  |
100;50;10;100 | 37°C |
| downstream 587 | [http://www.neb.com/nebecomm/products/productR3151.asp PvuII-HF] |  |
0;25;0;100 | 37°C |
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 10
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]