Part:BBa_K3117047:Design
scFv against GPA33 with SpyCatcher codon optimized for E.coli
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 54
Illegal AgeI site found at 519 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
By connecting the variable regions of the heavy and light chain of an anti-GPA33 antibody with a short, flexible GGGGS linker (BBa_K3117028), the scFv retains it's antigen-binding ability and is much smaller than a conventional antibody. The SpyCatcher attached to the scFv belongs to the SpyTag/SpyCatcher system, of which the sequence is derived of the FbaB protein in the bacteria Streptococcus pyogenes. Once it comes into contact with its corresponding other part, the SpyTag (BBa_K3117015), they bind covalently (Hatlem et al., 2019). This allows our part to be used in a modular manner in combination with other molecules carrying the SpyTag. The sequence contains a C-terminal His-Tag (BBa_K3117005) for easy purification and detection. Secretion of the protein into the periplasm is ensured by a pelB sequence (BBa_K3117012).
Source
The sequence of this part origins from the patent AU2018260975A1 [2].
References
1. Hatlem, D., Trunk, T., Linke, D., & Leo, J. C. (2019). Catching a SPY: Using the SpyCatcher-SpyTag and Related Systems for Labeling and Localizing Bacterial Proteins. International journal of molecular sciences, 20(9), 2129.
2. Bonvini, E., Moore, P. A., Li, J. C., Johnson, L. S., & Shah, K. (2017). U.S. Patent Application No. 15/313,765.