Composite

Part:BBa_K4392003:Design

Designed by: Vikram Venugopal   Group: iGEM22_IISER-Pune-India   (2022-10-01)


T7 overexpression part for acdS


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 39
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 950
    Illegal BamHI site found at 1098
    Illegal XhoI site found at 267
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 39
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 39
    Illegal NgoMIV site found at 1055
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 290
    Illegal BsaI.rc site found at 20


Design Notes

The T7 promoter, RBS, and terminator ensure compatibility with T7 RNA polymerase and provide good overexpression. The 6x His tags are present to help in protein purification through affinity column chromatography.


Source

This part was constructed through the restriction-free cloning of the acdS gene onto the pHis17 vector, which contained the T7 expression sequence for the parM 1 gene.

References