Composite

Part:BBa_K3778039:Design

Designed by: Yun Tsai   Group: iGEM21_CCU_Taiwan   (2021-09-06)


Thrombin releasable TAT48-57-6xHis-Cathepsin S releasable DPK-060


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 761
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 761
    Illegal NheI site found at 930
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 761
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 761
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 761
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This composite part contains T7 promoter, RBS, 6xHis, Enterokinase cleavage site, CBD, linker, Thrombin releasable TAT48-57, Cathepsin S releasable DPK-060, and T7 terminator. The T7 promoter, T7 terminator, and RBS are basic components for protein synthesis. The first 6xHis is applied for protein purification. Enterokinase site is responsible for the first 6xHis removal after purification. The second 6xHis is applied as a tag to examine whether Cathepsin S releasable DPK-060 get into the cells by immunostaining. CBD mediates the interaction with collagen and TAT48-57-Cathepsin S releasable-DPK 060 is the major effector for intra-bacterial clearance. DPK-060 is selected according to its resistance to Cathepsin S degradation.



Source

References