Coding

Part:BBa_K4825009:Design

Designed by: Ni Weizhao   Group: iGEM23_GreatBay-SCIE   (2023-10-11)


LEU1


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 643
    Illegal EcoRI site found at 1432
    Illegal PstI site found at 617
    Illegal PstI site found at 1112
    Illegal PstI site found at 1397
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 643
    Illegal EcoRI site found at 1432
    Illegal PstI site found at 617
    Illegal PstI site found at 1112
    Illegal PstI site found at 1397
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 643
    Illegal EcoRI site found at 1432
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 643
    Illegal EcoRI site found at 1432
    Illegal PstI site found at 617
    Illegal PstI site found at 1112
    Illegal PstI site found at 1397
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 643
    Illegal EcoRI site found at 1432
    Illegal PstI site found at 617
    Illegal PstI site found at 1112
    Illegal PstI site found at 1397
    Illegal NgoMIV site found at 160
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 1622


Design Notes

we engineered LEU1, LEU2, and LEU4 with three different signal peptide, which all lead them into mitochondria. Therefore, the concentration of enzymes that involve in 2-ketoacid elongation would become much higher in mitochondria, so that KIV will be convert to KIC before encountering KDC. In the end, KIC would be secreted out into cytosol and convert to Isopentanol


Source

Saccharomyces cerevisiae

References

Sarah K. Hammer, Yanfei Zhang, and José L. Avalos ACS Synthetic Biology 2020 9 (3), 546-555 DOI: 10.1021/acssynbio.9b00420