Composite

Part:BBa_K5159030

Designed by: Tianyue WU   Group: iGEM24_HKUST   (2024-10-02)


pTEFin-GapC-LIP2t-LEU2

This part is designed for expression of GapC gene under the strong promoter pTEF with intron in Yarrowia lipolytica. The pTEFin strong promoter is used as we aim to reach a high expression level for easier detection, and more importantly, to raise the flux of NADPH to compensate for its loss during DHA production in our project. 2 spacers were added before and after the LIP2 terminator to facilitate seamless assembly and provide a gap between the terminator and the next promoter to avoid interference. A LEU2 marker expression cassette is included for transformation selection in Yarrowia lipolytica Po1f strain with leucine auxotroph.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1947
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1947
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1947
    Illegal BglII site found at 4179
    Illegal BamHI site found at 1713
    Illegal XhoI site found at 2893
    Illegal XhoI site found at 2922
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1947
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1947
    Illegal NgoMIV site found at 2035
    Illegal AgeI site found at 3507
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 2553
    Illegal BsaI site found at 2836
    Illegal BsaI.rc site found at 2
    Illegal BsaI.rc site found at 2751
    Illegal SapI site found at 577
    Illegal SapI site found at 3304


[edit]
Categories
Parameters
None