Composite

Part:BBa_K5159023:Design

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


pTEF-ORFC-LIP2t-LEU2


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 5451
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 5451
    Illegal NotI site found at 2885
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 5451
    Illegal BglII site found at 7683
    Illegal BamHI site found at 1501
    Illegal BamHI site found at 5217
    Illegal XhoI site found at 631
    Illegal XhoI site found at 997
    Illegal XhoI site found at 1444
    Illegal XhoI site found at 3447
    Illegal XhoI site found at 3514
    Illegal XhoI site found at 3799
    Illegal XhoI site found at 3874
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 5451
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 5451
    Illegal NgoMIV site found at 1120
    Illegal NgoMIV site found at 1150
    Illegal NgoMIV site found at 2242
    Illegal NgoMIV site found at 2788
    Illegal NgoMIV site found at 3730
    Illegal NgoMIV site found at 3763
    Illegal NgoMIV site found at 3976
    Illegal AgeI site found at 7011
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 2853
    Illegal BsaI site found at 4035
    Illegal BsaI site found at 4681
    Illegal BsaI site found at 6057
    Illegal BsaI site found at 6340
    Illegal BsaI.rc site found at 2
    Illegal BsaI.rc site found at 991
    Illegal BsaI.rc site found at 3652
    Illegal BsaI.rc site found at 4657
    Illegal BsaI.rc site found at 4993
    Illegal BsaI.rc site found at 6255
    Illegal SapI site found at 6808
    Illegal SapI.rc site found at 2994


Design Notes

ORF_C 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