Composite

Part:BBa_K2842666:Design

Designed by: Catherine Fan, Stefanie Santamaria   Group: iGEM18_UCL   (2018-10-01)


Golden Gate compatible system with blue-white screening


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 site found at 647
    Illegal BsaI.rc site found at 28


Design Notes

The initial version of BioBrick2.0 contained a PvuII site which is advised against in RFC12. Within LacZa a silent mutation T to C mutation was introduced to maintain the underlying Ser. For the second PvuII site, a C to G mutation was introduced furthest from the -35/-10 sites.


Source

This construct was designed on Benchling and synthesised by IDT.


References

Vladimir P, Ong JL, Kucera RB, Langhorst BW, Bilotti K, Pryor JM, et al. Optimization of Golden Gate assembly through application of ligation sequence-dependent fidelity and bias profiling. bioRxiv [Internet]. 2018 May 15 [cited 2018 Oct 16];322297. Available from: https://www.biorxiv.org/content/early/2018/05/15/322297