Project

Part:BBa_K404251

Designed by: Freiburg Bioware 2010   Group: iGEM10_Freiburg_Bioware   (2010-10-14)
Revision as of 02:48, 28 October 2010 by Jessica (Talk | contribs)

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 25
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


[edit]
Categories
//chassis/eukaryote/human
//viral_vectors
//viral_vectors/aav
//viral_vectors/aav/capsid_coding
Parameters
None