Composite

Part:BBa_K5477026:Design

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


pRET2-NCOA - receptor module


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1785
    Illegal EcoRI site found at 2407
    Illegal SpeI site found at 1234
    Illegal SpeI site found at 1535
    Illegal SpeI site found at 2602
    Illegal PstI site found at 1294
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1785
    Illegal EcoRI site found at 2407
    Illegal NheI site found at 182
    Illegal NheI site found at 3547
    Illegal SpeI site found at 1234
    Illegal SpeI site found at 1535
    Illegal SpeI site found at 2602
    Illegal PstI site found at 1294
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1785
    Illegal EcoRI site found at 2407
    Illegal BamHI site found at 3049
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1785
    Illegal EcoRI site found at 2407
    Illegal SpeI site found at 1234
    Illegal SpeI site found at 1535
    Illegal SpeI site found at 2602
    Illegal PstI site found at 1294
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1785
    Illegal EcoRI site found at 2407
    Illegal SpeI site found at 1234
    Illegal SpeI site found at 1535
    Illegal SpeI site found at 2602
    Illegal PstI site found at 1294
    Illegal AgeI site found at 37
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This module supports the AhR-mediated response to toxic ligands like PAHs, dioxins and PCBs by binding to the XRE() in our reporter module facilitating the transcription of NanoLuc (). The composite part was cloned using the method of USER-cloning into YCp-W. YCp-W is a centromeric plasmid used in yeast that carries a TRP1 marker, allowing for selection in tryptophan auxotrophic yeast strains. Like other CEN plasmids, YCp-W contains a CEN sequence, ensuring that it replicates and segregates similarly to yeast chromosomes. This low-copy plasmid (typically maintained at one to two copies per cell) provides stable maintenance within yeast cells.



Source

Homo sapiens and Saccharomyces cerevisiae

References