Plasmid

Part:BBa_K4212042:Design

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


ChiS9


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1110
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1110
    Illegal NheI site found at 206
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1110
    Illegal BglII site found at 101
    Illegal BamHI site found at 1095
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1110
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1110
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

We intended to construct a Level 1 Golden Gate Plasmid by combining the native promoter of CotG, the CotG-chitinase fusion protein. We aimed to compared the transcription effectiveness of the native promoter with the hyperspank promoter, and we would select the more suitable one for future Golden Gate assembly. We added a flexible linker to this construct to test if it works better than the construct without the linker. We also tried out different linkers to compare their effectiveness. We would then change our construct design and the media we use accordingly. 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