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Revision as of 17:16, 26 October 2010
ViralBrick-587KO-BAP
The Biotinylation Acceptor peptide motif & HSPG KO, 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]