Coding

Part:BBa_K4061020:Design

Designed by: Diya Agrawal   Group: iGEM21_HKUST   (2021-09-30)


Engineered EnvZ (facilitates scaffolding)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NotI site found at 1396
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 942
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 1546


Design Notes

The SH3 ligand that is present within the protein CDS was supposed to have a Kd higher than that present on the scaffold. Moreover, we needed to model the protein using SWISS-MODEL to be certain of the helix-four-bindle region before placing our SH3 ligand auto-inhibition sequence there.



Source

The EnvZ sequence was obtained from UniProt while the SH3 binding domain and ligand were inspired by paper by Whitaker, W.R. et al (2014) titled "Engineering robust control of two-component system phosphotransfer using modular scaffolds".

References