Composite

Part:BBa_K5159024:Design

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


pTEF-PPT-LIP2t-LEU2


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


Design Notes

PPTase is expressed under strong constitutive promoter pTEFin as we aim to reach a high expression level and facilitate the detection before applying any inducible promoters for production stage control. A 30 bp spacers were added before the LIP2 terminator to facilitate seamless assembly by offering a suitable GC content range for designing overhang primers. The 200 bp spacers added after the LIP2 terminator provided a gap between the terminator and the next promoter to avoid interference.



Source

Please refer to the sources of each subparts.

References