Composite

Part:BBa_K4825058:Design

Designed by: Ni Weizhao   Group: iGEM23_GreatBay-SCIE   (2023-10-12)


pPGK1-ERG20ww-tPGK1-pTDH3-t48PS-tTDH1


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1105
    Illegal BglII site found at 1350
    Illegal BglII site found at 4024
    Illegal BamHI site found at 2187
    Illegal BamHI site found at 4861
    Illegal XhoI site found at 5561
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 3
    Illegal BsaI.rc site found at 1115
    Illegal BsaI.rc site found at 1822
    Illegal BsaI.rc site found at 4496


Design Notes

Geranyl pyrophosphate(GPP) is precursor of pinene, which is our desired product. Therefore a higher yield of GPP is desirable, while the basic level of FPP is crucial for yeast survival. A method is designed to increase production of GPP while keeping FPP at a low level. It is to introduce 2 mutation F96W and N127W in ERG20, creating ERG20ww. The mutation in ERG20ww enables the reduction in FPP synthesis(from GPP), while GPP synthesis is not influenced. As a result, less GPP will be transformed to FPP, and a higher yield of pinene could be achieved. "1. The 48-locus site at the N terminal of PtPS was truncated to produce t48PS, which can increase the efficiency of pinene production 2. PS is linked to ERG20ww with a flexible linker (GGGGSGGGGS) to increase the production of pinene from GPP, and reduce the GPP allocated to produce FPP to increase the yield of target product."


Source

Saccharomyces cerevisiae, Pinus taeda

References