Composite

Part:BBa_K5159024

Designed by: Tianyue WU   Group: iGEM24_HKUST   (2024-09-30)


pTEF-PPT-LIP2t-LEU2

This part is designed for expression of phosphopantetheinyl transferase (PPT) under the strong promoter pTEF with intron, one of the four constructs for introducing the DHA-producing PUFA synthase pathway into Yarrowia lipolytica. It is expressed under this strong promoter as we aim to reach a high PPTase expression level to activate the DHA synthesis pathway, leading to a high DHA yield. 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 1758
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1758
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1758
    Illegal BglII site found at 3990
    Illegal BamHI site found at 853
    Illegal BamHI site found at 1524
    Illegal XhoI site found at 2704
    Illegal XhoI site found at 2733
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1758
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1758
    Illegal NgoMIV site found at 550
    Illegal NgoMIV site found at 1846
    Illegal AgeI site found at 3318
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 2364
    Illegal BsaI site found at 2647
    Illegal BsaI.rc site found at 2
    Illegal BsaI.rc site found at 2562
    Illegal SapI site found at 3115
    Illegal SapI.rc site found at 652
    Illegal SapI.rc site found at 1267


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