Device

Part:BBa_K2940014:Design

Designed by: Jakub Chromy   Group: iGEM19_Edinburgh_OG   (2019-10-16)


XylR coding device-sfGFP_optimism coding device with Pu


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 2446
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 2101
    Illegal XhoI site found at 3018
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 329
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The part is identical to the BBa_K2023015 part developed by Ionis Paris iGEM team 2016 [1], except for the originally used wild-type GFP (BBa_E0040) reporter gene being replaced by codon-optimized superfolder GFP (BBa_K2541400).

The codon-optimized superfolder GFP variant was used as the reporter in an attempt to have a very sensitive starting biosensor genetic circuit for the subsequent directed evolution and screenings for the biosensors with new substrate specificities. However, the use of this strong GFP variant resulted in strongly constitutively fluorescent E. coli cells impeding the function of the biosensor.

Source

Original XylR:GFP biosensor genetic circuit (BBa_K2023015) with sfGFP variant (BBa_K2541400) incorporated.

References

[1] http://2016.igem.org/Team:Ionis_Paris