Coding

Part:BBa_K2721024:Design

Designed by: Yunqian Li   Group: iGEM18_ZJU-China   (2018-10-05)


RFP with SdyTag and SpyCatcher


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 784
    Illegal AgeI site found at 600
    Illegal AgeI site found at 712
  • 1000
    COMPATIBLE WITH RFC[1000]


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 RFP is a reporter.

We have mutated some illegal enzyme sites through synonymous substitutions to make it compatible with RFC[10].

Source

SpyCatcher is derived from Streptococcus pyogenes fibronectin-binding protein. SdyTag is derived from a related fibronectin-binding protein in Streptococcus dysgalactiae.

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. Tan LL, Hoon SS, Wong FT. Kinetic Controlled Tag-Catcher Interactions for Directed Covalent Protein Assembly. PLoS ONE 11(10): e0165074. https://doi.org/10.1371/journal.pone.0165074

3. P Hengen. Purification of His-Tag fusion proteins from Escherichia coli. Trends in Biochemical Sciences, 1995,20(7): 285-286.