Composite
OmpR-RFR-N

Part:BBa_K2845037:Design

Designed by: Jan Lukas Krüsemann, Antonio Martínez Brotons, Eline Bijman   Group: iGEM18_ETH_Zurich   (2018-10-10)


OmpR fused the N-terminus of split EPIC luciferase


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

OmpR is fused to the N-terminus of split EPIC Luciferase. OmpR is part of the natural response system from E. coli reacting to osmotic changes and inducing adaptive changes in the membrane. Only when phosphorylated by EnvZ, it has an increased binding affinity to its binding site and activates the expression of membrane proteins. This biobrick is designed to be used in a system together with a fusion protein consisting of the C-terminus of the split EPIC luciferase fused to TetR and the binding sites of OmpR and TetR placed next to each other on a high copy plasmid. Since TetR is constitutively binding to its binding site, the two luciferase halves are only brought into proximity when OmpR gets phosphorylated and binds to its binding site. This allows the visualization of transcription factor binding by circumventing the time-consuming step of transcription and translation.