Coding

Part:BBa_K3515006:Design

Designed by: Kody Klupt, Sono Khan   Group: iGEM20_Queens_Canada   (2020-05-25)
Revision as of 13:44, 25 May 2020 by Kody klupt (Talk | contribs)


Synechococcus Phosphate Binding Protein with cysteine modification(s) and FRET to monitor phosphate


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1539
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1129
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Added a Cysteine for immobilization (D256C) opposite to the active site region. Removed C135A and C189S to ensure only one cysteine would bind to an immobilization linker arm. Amino acid substitution considerations were made using an intensive BLAST search to ensure conservation. A T22A modification was made based on previous evidence of this substitution permitting phosphate binding in the physiological range. No modifications were made to the FRET acceptor or donor.


Source

The source of this part is its sequence retrieved from the European Nucleotide Archive (BX569691.1) along with our own modifications.

References

Gu, H., Lalonde, S., Okumoto, S., Looger, L.L., Scharff-Poulsen, A.M., Grossman, A.R., Kossmann, J., Jakobsen, I. and Frommer, W.B., 2006. A novel analytical method for in vivo phosphate tracking. FEBS letters, 580(25), pp.5885-5893.