Composite
Part:BBa_K5477023:Design
Designed by: Kate Malana Escobar Group: iGEM24_UCopenhagen (2024-09-26)
pSTE12-AhR - receptor module
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 927
Illegal XbaI site found at 55 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 927
Illegal NheI site found at 732 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 927
Illegal BglII site found at 1742 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 927
Illegal XbaI site found at 55 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 927
Illegal XbaI site found at 55 - 1000COMPATIBLE 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