Difference between revisions of "Part:BBa K3117047:Design"
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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.
 
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.
  
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2.[2]: Bonvini, E., Moore, P. A., Li, J. C., Johnson, L. S., & Shah, K. (2017). U.S. Patent Application No. 15/313,765.
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2. Bonvini, E., Moore, P. A., Li, J. C., Johnson, L. S., & Shah, K. (2017). U.S. Patent Application No. 15/313,765.

Latest revision as of 02:13, 22 October 2019


scFv against GPA33 with SpyCatcher codon optimized for E.coli


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 54
    Illegal AgeI site found at 519
  • 1000
    COMPATIBLE 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.