Composite

Part:BBa_K5159022:Design

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


pTEF-ORFB-LIP2t-LEU2


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 7122
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 7122
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 7122
    Illegal BglII site found at 9354
    Illegal BamHI site found at 6888
    Illegal XhoI site found at 2293
    Illegal XhoI site found at 2539
    Illegal XhoI site found at 2854
    Illegal XhoI site found at 3445
    Illegal XhoI site found at 3817
    Illegal XhoI site found at 4935
    Illegal XhoI site found at 5563
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 7122
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 7122
    Illegal NgoMIV site found at 1648
    Illegal NgoMIV site found at 4492
    Illegal NgoMIV site found at 5392
    Illegal NgoMIV site found at 5425
    Illegal NgoMIV site found at 5965
    Illegal NgoMIV site found at 7210
    Illegal AgeI site found at 3019
    Illegal AgeI site found at 8682
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1804
    Illegal BsaI site found at 3994
    Illegal BsaI site found at 5697
    Illegal BsaI site found at 7728
    Illegal BsaI site found at 8011
    Illegal BsaI.rc site found at 2
    Illegal BsaI.rc site found at 1726
    Illegal BsaI.rc site found at 1777
    Illegal BsaI.rc site found at 2542
    Illegal BsaI.rc site found at 3460
    Illegal BsaI.rc site found at 4537
    Illegal BsaI.rc site found at 5002
    Illegal SapI site found at 8479


Design Notes

ORF_B 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