Composite

Part:BBa_K5477038

Designed by: Kate Malana Escobar   Group: iGEM24_UCopenhagen   (2024-09-28)
Revision as of 20:27, 28 September 2024 by Kateesc1700 (Talk | contribs)


CYP3A4-pGAL1/10-POR detox module against a wide array of contaminants

In our system, the CYP3A4-pGAL1/10-POR composite part combines the CYP3A4 enzyme and cytochrome P450 oxidoreductase (POR) to form a highly efficient detoxification unit. The pGAL1/10 bidirectional promoter drives the expression of both CYP3A4 and POR simultaneously but in opposite directions. CYP3A4, a key phase I enzyme, is responsible for oxidizing around 50% of all clinically used drugs and environmental toxins. POR provides the electrons required for these oxidation reactions by transferring electrons from NADPH to CYP3A4, ensuring efficient metabolism of harmful xenobiotics and pharmaceuticals.

This 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.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 1483
    Illegal PstI site found at 2486
    Illegal PstI site found at 2683
    Illegal PstI site found at 3067
    Illegal PstI site found at 3507
    Illegal PstI site found at 3567
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 2486
    Illegal PstI site found at 2683
    Illegal PstI site found at 3067
    Illegal PstI site found at 3507
    Illegal PstI site found at 3567
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 1483
    Illegal PstI site found at 2486
    Illegal PstI site found at 2683
    Illegal PstI site found at 3067
    Illegal PstI site found at 3507
    Illegal PstI site found at 3567
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 1483
    Illegal PstI site found at 2486
    Illegal PstI site found at 2683
    Illegal PstI site found at 3067
    Illegal PstI site found at 3507
    Illegal PstI site found at 3567
    Illegal NgoMIV site found at 2889
    Illegal NgoMIV site found at 3008
    Illegal AgeI site found at 1894
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None