Coding

Part:BBa_K3515003:Design

Designed by: Kody Klupt, Sono Khan   Group: iGEM20_Queens_Canada   (2020-05-22)


Troponin C, a calcium binding protein with cysteine modification(s) to bind to a biosensor.


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 AgeI site found at 376
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Added a cysteine for immobilization (K93C) opposite to the active site region for linker arm immobilization and removed a cysteine previously existing (C101A). Amino acid substitution considerations were made using an intensive BLAST search to ensure conservation. N109D, D111N, N145D, and D147N modifications were made based on previous evidence of these substitutions permitting selective calcium binding in the physiological range and removing the ability of magnesium to bind competitively.


Source

The source of this part is its sequence retrieved from the European Nucleotide Archive (M19027.1) along with our own modifications.

References

Mank, M., Reiff, D.F., Heim, N., Friedrich, M.W., Borst, A. and Griesbeck, O., 2006. A FRET-based calcium biosensor with fast signal kinetics and high fluorescence change. Biophysical journal, 90(5), pp.1790-1796