Composite

Part:BBa_K4164028

Designed by: Bo Cheng   Group: iGEM22_NAU-CHINA   (2022-10-09)


pCadBA-GFP

This composite part contains the pCadBA promoter (BBa_K3425101) and green fluorescent protein (BBa_E0040). When the CadC domains dimerize, the downstream CadBA promoter will drive green fluorescent protein (GFP) expression.

We recombined pCadBA promoter (BBa_K3425101) and green fluorescent protein (BBa_E0040) by homologous recombination.

With the system above, we selected CadC, promoter CadBA, GFP as our report device. When FXR and RXR dimerize, the linked CadC will be in close proximity and the dimerization occurs, activating promoter CadBA to express GFP.

Finally, we chose FXR and RXR as bile acid sensing receptors, GFP as a signaling device and E. coli BL21 as our chassis organism, hoping to demonstrate the feasibility of our project.

Fig.1. Gene circuit based on GFP

Fig.2. Fluorescence intensity/OD using a gradient of CDCA concentrations based on ddRFP.

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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 800


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