Plasmid

Part:BBa_K5371204:Design

Designed by: Haoyang Yu   Group: iGEM24_iZJU-China   (2024-09-29)


prs42b-erg9


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 2934
    Illegal XbaI site found at 193
    Illegal XbaI site found at 2692
    Illegal XbaI site found at 7554
    Illegal SpeI site found at 3593
    Illegal PstI site found at 408
    Illegal PstI site found at 872
    Illegal PstI site found at 3237
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 2934
    Illegal SpeI site found at 3593
    Illegal PstI site found at 408
    Illegal PstI site found at 872
    Illegal PstI site found at 3237
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 2934
    Illegal BglII site found at 198
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 2934
    Illegal XbaI site found at 193
    Illegal XbaI site found at 2692
    Illegal XbaI site found at 7554
    Illegal SpeI site found at 3593
    Illegal PstI site found at 408
    Illegal PstI site found at 872
    Illegal PstI site found at 3237
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 2934
    Illegal XbaI site found at 193
    Illegal XbaI site found at 2692
    Illegal XbaI site found at 7554
    Illegal SpeI site found at 3593
    Illegal PstI site found at 408
    Illegal PstI site found at 872
    Illegal PstI site found at 3237
    Illegal NgoMIV site found at 683
    Illegal NgoMIV site found at 1813
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 3541
    Illegal BsaI.rc site found at 2313
    Illegal BsaI.rc site found at 2696
    Illegal BsaI.rc site found at 5724
    Illegal SapI site found at 4641


Design Notes

Homologous regions were chosen 100bp around the original ERG9 promoter for better integration efficiency. HXT1 promoter was chosen for knocking down ERG9 as it is a weaker promoter, compared to the original one, and also for the actual culturing environment, which will be galactose dominant and further suppress expression level of ERG9.


Source

prs42b-erg9 was constructed via Gibson Assembly, with segments mainly amplified from Saccharomyces cerevisiae.

References