Composite

Part:BBa_K5477023:Design

Designed by: Kate Malana Escobar   Group: iGEM24_UCopenhagen   (2024-09-26)


pSTE12-AhR - receptor module


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 927
    Illegal XbaI site found at 55
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 927
    Illegal NheI site found at 732
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 927
    Illegal BglII site found at 1742
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 927
    Illegal XbaI site found at 55
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 927
    Illegal XbaI site found at 55
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

In our pSTE12-AhR receptor module, the pSTE12 promoter drives the expression of AhR. Once AhR is expressed and activated by ligands like PAHs or PCBs, it binds to the XRE sequences in a reporter module, triggering the expression of NanoLuc. The composite part was cloned using the method of USER-cloning into YCp-L. The YCp-L plasmid is a centromeric plasmid that carries a LEU2 marker for leucine selection in yeast. Centromeric plasmids like YCp-L are low-copy vectors, typically maintained at one to two copies per cell due to the presence of a CEN sequence.



Source

Homo sapiens and Saccharomyces cerevisiae

References