Plasmid

Part:BBa_K4212038:Design

Designed by: Fontaine Gibbs   Group: iGEM22_Imperial_College_London   (2022-09-30)


ChiS5


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1723
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1723
    Illegal NheI site found at 1011
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1723
    Illegal BglII site found at 1066
    Illegal BamHI site found at 1708
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1723
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1723
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This part consists the fusion protein consisted of CotZ and ChiS, in which we combined these parts into level 1 Golden Gate Construct. Our dry lab team predicted the functional model of this chimera and our wet lab intended to test the effectiveness of this model. We intended to compare the fungal-killing efficiency of CotZ and CotG fusion protein. Promoter hyper-spank is an inducible promoter, which was designed for high-level protein expression in B.subtilis and it was previously used by other iGEM teams.Our dry lab team predicted 2 possible chimeras, with either CotG or CotZ attached to the chitinase. Our wet lab team attempted to verify that the CotG chimera works better than the CotZ one. Promoter hyper-spank is an inducible promoter, which was designed for high-level protein expression in B.subtilis and it was previously used by other iGEM teams. The addition of Isopropyl β-D-1-thiogalactopyranoside (IPTG) can induce gene expression under the control of the promoter hyperspank. tL3S2P21 is a strong terminator, which helps to increase the production of desired protein.


Source

Synthetic construct.

References

https://parts.igem.org/Part:BBa_K143015