Part:BBa_K4701204

SpyCatcher002 (with C-terminus linker)

The FbaB protein found in S. pyogenes contains a CnaB2 domain with a highly stable isopeptide bond [1]. This domain has been split into two fragments and engineered, the larger of the two called SpyCatcher spontaneously forms an isopeptide bond with the smaller fragment called SpyTag. This allows for the creation of fusion proteins. SpyCatcher002 is an updated version that more effectively forms these spontaneous bonds [2]. The part includes a Gly/Ser-linker recommended by the authors of the system [3] on the C-terminus to separate SpyCatcher002 from the protein of interest, when SpyCatcher002 is used in the N-terminus of a protein.

References

[1] Zakeri, B., et al. (2012). Peptide tag forming a rapid covalent bond to a protein, through engineering a bacterial adhesin. https://doi.org/10.1073/pnas.1115485109

[2] Keeble, A. H., et al. (2017). Evolving Accelerated Amidation by SpyTag/SpyCatcher to Analyze Membrane Dynamics. Angewandte Chemie, 129(52), 16748–16752. https://doi.org/10.1002/ange.201707623

[3] Keeble, A. H., & Howarth, M. (2019). Insider information on successful covalent protein coupling with help from SpyBank. In Methods in Enzymology (Vol. 617, pp. 443–461). Academic Press Inc. https://doi.org/10.1016/bs.mie.2018.12.010

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]