Difference between revisions of "Part:BBa K404205"

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:

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]

@@ Line 1: / Line 1: @@
 __NOTOC__
 <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>
@@ Line 10: / Line 29: @@
 <div style="width:965px; height:400px; ">
 <div style="float:left; width:460px; height:auto;  margin: 0px 5px 0px 5px; text-align:justify;">
-The BAP (Biotinylation Acceptor Peptide) that we included in our Virus Construction Kit is a 15 amino acid long peptide identified by Schatz J.,  1993 in an library screening approach  and published under the number #85. 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). <br>
-The purified BirA biotin ligase that was kindly provided from Avidity.com
+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).
-<h3>References:</h3>
 </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"
-height="auto"/></div></div><br><br><br><br>
+height="auto"/></div></div><br>
 </html>