Composite

Part:BBa_K3786001:Design

Designed by: Camila Ballenghien   Group: iGEM21_Paris_Bettencourt   (2021-10-21)


GFP expression controlled by Arabinose operon


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1205
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1144
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 979
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1867
    Illegal SapI site found at 961


Design Notes

While designing this composite part, we used golden gate cloning therefore we had to verify that none of the sequences contained BsaI cut sites within them. This wasn't a problem for the GFP and the terminator as they came from the CIDAR MoClo Kit and the parts are optimized for Golden gate. However, for the Arabinose operon this was important to take into account or else Golden gate wouldn't have been a possibility.


Source

Arabinose operon and the pET ribosome binding site was amplified from a plasmid pBAD-pR (Song et al., 2019), the GFP sequence, the terminator (B0015) and the backbone are from the CIDAR MoClo Toolkit.

References

References:

Song, Y., Cartron, M. L., Jackson, P. J., Davison, P. A., Dickman, M. J., Zhu, D., Huang, W. E., & Hunter, C. N. (2019). Proteorhodopsin Overproduction Enhances the Long-Term Viability of Escherichia coli. Applied and Environmental Microbiology, 86(1). https://doi.org/10.1128/aem.02087-19