Other
T-10-O

Part:BBa_K2845025:Design

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


TetR binding site next to OmpR binding site separated by 10 bases.


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

For circumventing the gene transcriptional lag, this biobrick uses DNA as a scaffold to visualize transcription factor binding. The OmpR transcription factor from E. coli's natural EnvZ/OmpR two component system binds to pOmpC. It can be fused to one half of a split luciferase while the other split luciferase half is fused to TetR. On a high-copy plasmid, the binding sites for OmpR and TetR are placed next to each other. While TetR constitutively binds to DNA, OmpR needs to be phosphorylated by EnvZ to get a higher binding affinity to its binding site. Upon binding of OmpR, the two split luciferase halves are brought into proximity, allowing them to re-assemble and to emit light, thereby increasing biosensor output.