Difference between revisions of "Part:BBa K4390065"

Revision as of 15:57, 12 October 2022

His-tagged SpyTagged sfGFP expression construct

This part is not compatible with BioBrick RFC10 assembly but is compatible with the iGEM Type IIS Part standard which is also accepted by iGEM.

Usage and Biology

SpyTag is a 13 amino acid-long protein that spontaneously reacts with the SpyCatcher protein (12.3 kDa). Together they form an intermolecular isopeptide bond. This provides an alternative to His-tag purification techniques. Insertion of either the SpyTag or the SpyCatcher sequence into a recombinant protein will allow its purification.

By adding a SpyTag to sfGFP, this can be used to see how efficient the SpyCatcher is at binding the SpyTag bound to the sfGFP (as a control, to see the binding efficiency of binding to the tagged proteins in general).

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 11
Illegal NheI site found at 34
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 7: / Line 7: @@
 ===Usage and Biology===
 SpyTag is a 13 amino acid-long protein that spontaneously reacts with the SpyCatcher protein (12.3 kDa). Together they form an intermolecular isopeptide bond. This provides an alternative to His-tag purification techniques. Insertion of either the SpyTag or the SpyCatcher sequence into a recombinant protein will allow its purification.
+By adding a SpyTag to sfGFP, this can be used to see how efficient the SpyCatcher is at binding the SpyTag bound to the sfGFP (as a control, to see the binding efficiency of binding to the tagged proteins in general).
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