Coding
Part:BBa_K2721023:Design
Designed by: Yunqian Li Group: iGEM18_ZJU-China (2018-10-04)
SpyTag with GFP
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 689
Design Notes
The tag-catcher pairs are ideal for binding, immobilization and creating new kinds of protein architectures. Our team use them as scaffold of protein assembly. Here GFP is a reporter.
We have mutated some illegal enzyme sites through synonymous substitutions to make it compatible with RFC[10].
Source
SpyTag is derived from Streptococcus pyogenes fibronectin-binding protein.
References
1. Zakeri B, Fierer JO, Celik E, Chittock EC, Schwarz-Linek U, Moy VT, Howarth M. Peptide tag forming a rapid covalent bond to a protein, through engineering a bacterial adhesin. Proc Natl Acad Sci U S A. 2012 Mar 20;109(12):E690-7.
2. J Fang, WB Zhang. Genetically Encoded Peptide-protein Reactive Pairs. Acta Polymerica Sinica 2018 Jan (4):429-444.