Part:BBa_K2721024:Design

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.