Composite

Part:BBa_K5159021:Design

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


pTEF-ORFA-LIP2t-LEU2


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 9675
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 9675
    Illegal NotI site found at 7250
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 9675
    Illegal BglII site found at 737
    Illegal BglII site found at 2540
    Illegal BglII site found at 2600
    Illegal BglII site found at 11907
    Illegal BamHI site found at 9441
    Illegal XhoI site found at 1945
    Illegal XhoI site found at 3976
    Illegal XhoI site found at 4219
    Illegal XhoI site found at 4303
    Illegal XhoI site found at 4636
    Illegal XhoI site found at 4872
    Illegal XhoI site found at 4885
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 9675
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 9675
    Illegal NgoMIV site found at 652
    Illegal NgoMIV site found at 877
    Illegal NgoMIV site found at 1477
    Illegal NgoMIV site found at 1897
    Illegal NgoMIV site found at 2614
    Illegal NgoMIV site found at 2638
    Illegal NgoMIV site found at 6112
    Illegal AgeI site found at 1246
    Illegal AgeI site found at 1585
    Illegal AgeI site found at 8083
    Illegal AgeI site found at 11235
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1723
    Illegal BsaI site found at 3012
    Illegal BsaI site found at 7021
    Illegal BsaI site found at 7695
    Illegal BsaI site found at 8086
    Illegal BsaI site found at 10281
    Illegal BsaI site found at 10564
    Illegal BsaI.rc site found at 2
    Illegal BsaI.rc site found at 640
    Illegal BsaI.rc site found at 2092
    Illegal BsaI.rc site found at 3037
    Illegal BsaI.rc site found at 3067
    Illegal BsaI.rc site found at 3217
    Illegal BsaI.rc site found at 3946
    Illegal SapI site found at 2340
    Illegal SapI site found at 3045
    Illegal SapI site found at 8613
    Illegal SapI site found at 11032


Design Notes

ORF_A 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