Composite

Part:BBa_K5477025:Design

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


pRET2 - ARNT receptor module


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 810
    Illegal EcoRI site found at 1916
    Illegal SpeI site found at 1760
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 810
    Illegal EcoRI site found at 1916
    Illegal NheI site found at 182
    Illegal SpeI site found at 1760
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 810
    Illegal EcoRI site found at 1916
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 810
    Illegal EcoRI site found at 1916
    Illegal SpeI site found at 1760
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 810
    Illegal EcoRI site found at 1916
    Illegal SpeI site found at 1760
    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-H. YCp-H is a centromeric plasmid used in yeast that includes a HIS3 marker, allowing for selection in histidine auxotrophic yeast strains. Like other CEN plasmids, YCp-H contains a CEN sequence, ensuring that the plasmid replicates and segregates similarly to yeast chromosomes. This results in a low copy number (typically one to two copies per cell), providing stable maintenance of the plasmid.


Source

Homo sapiens and Saccharomyces cerevisiae

References