Reporter

Part:BBa_K2450501:Design

Designed by: Zoe Ford   Group: iGEM17_Oxford   (2017-10-17)


SpyCatcher sfGFP quencher


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 4
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 397
    Illegal SapI.rc site found at 1291


Design Notes

The TorA leader sequence must be at the N terminal end.

The part has been designed so that the sfGFP stays attached to the membrane when the TEV protease cleaves the quencher, and the dark quencher is at the C terminus. This allows the quencher to be removed during PCR to produce a protein without the quencher. This part without the fluorophore can be used to compare the increase in fluorescence against, when the quencher is cleaved off.


Source

tba

References