Plasmid

Part:BBa_K4212047:Design

Designed by: Yuancheng Ding   Group: iGEM22_Imperial_College_London   (2022-09-30)


SDP4


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1743


Design Notes

GFPmut3 is a basic fluorescent protein, which can be used to differentiate the bacteria colonies which successfully take in the cloning construct. tL3S2P21 is a strong terminator, which helps to increase the production of desired 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. The addition of Isopropyl β-D-1-thiogalactopyranoside (IPTG) can induce gene expression under the control of the promoter hyperspank. We included it in our self-digesting plasmid construct to help us differentiate the bacteria colonies with success integration of the desired construct.


Source

Synthetic construct.

References

[1] Franke, G.C., Dobinsky, S., Mack, D., Wang, C.-J., Sobottka, I., Christner, M., Knobloch, J.K.-M., Horstkotte, M.A., Aepfelbacher, M. & Rohde, H. (2007) Expression and functional characterization of gfpmut3.1 and its unstable variants in Staphylococcus epidermidis. Journal of Microbiological Methods. 71 (2), 123–132. doi:10.1016/j.mimet.2007.08.015. [2] https://parts.igem.org/Part:BBa_K143015