Coding

Part:BBa_K4685019

Designed by: Kasandra Nicole Balzaretti   Group: iGEM23_UNILausanne   (2023-10-10)


SpyTag protein

This basic part, includes the gene for the SpyTag protein attached to a linker. This part is a subunit of the CsgA-SpyTag complex(BBa_K4685021). The CsgA-SpyTag complex is capable of interacting with the SpyCatcher protein fused to the Rubber-binding protein(BBa_K4685000).

The SpyTag/SpyCatcher system allows an irreversible conjugation of proteins:the peptide SpyTag reacts with the protein SpyCacther and forms an intramolecular isopeptide bond between the two proteins. We decided to leverage this key-lock mechanism by combining the SpyCatcher with the Rubber-binding protein and the SpyTag with the csgA subunit. Upon reaction between the two parts, the Curli fibers would be decorated with the Rubber-binding domain This composite part is different from BBa_K2684006, as it the sequence for the linker and SpyTag protein is different

  rubber-domain.png
  

Figure 1: Schematic representation of the SpyCatcher/SpyTag system applied to our project. The SpyTag protein, located on the surface of Curli fiber, will interact with the SpyCatcher protein that is bound to the Rubber-binding protein.


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]


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