Regulatory

Part:BBa_K4897014

Designed by: Hejia Dong   Group: iGEM23_BS-United-China   (2023-10-06)


LUXI promoter and EL222 binding sequence

What is it?

This part allows the gene expression during a dark environment. Aiming to develop this bule-light-activated suicide system, where our engineering E. coli will be killed during the absence of blue light, namely in dark or at night, we equipped our engineering bacteria with a special part to produce EL222, a light-regulated DNA-binding protein. This protein can harness blue light to drive the reorientation of its Light-Oxygen-Voltage (LOV) sensory and Helix-Turn-Helix (HTH) effector domains, resulting in its self-dimerization and thus binding to the EL222 binding region located between −35 (TTGACA) and −10 (TATAAT) region of the luxI promoter.

Usage and Biology

With this part built into our blue light suicide system, once blue light is undetectable by EL222, which means a dark environment is provided at night, the formation of EL222 dimers is terminated and the RNAP is allowed to bind back to the -35 to -10 region of the luxI promoter. This thereby activates the expression of the two types of CAS9 (CAS9-A and CAS9-R) and their sgRNA sequence (sgRNA-A and sgRNA-R). This allows our target bacteria to be killed and ensures the safety of envioronment.


Sequence and Features


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]


[edit]
Categories
Parameters
None