Coding

Part:BBa_K4825011:Design

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


LEU4^S547∆


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 397
    Illegal XbaI site found at 511
    Illegal SpeI site found at 552
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 75
    Illegal SpeI site found at 552
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1282
    Illegal BamHI site found at 228
    Illegal BamHI site found at 1194
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 397
    Illegal XbaI site found at 511
    Illegal SpeI site found at 552
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 397
    Illegal XbaI site found at 511
    Illegal SpeI site found at 552
  • 1000
    COMPATIBLE WITH RFC[1000]


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