RBS

Part:BBa_K3059620:Design

Designed by: Julia Urban   Group: iGEM19_William_and_Mary   (2019-10-16)


pre-csgE RBS


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
    COMPATIBLE WITH RFC[1000]


Design Notes

When designing this part, we were investigating a way to combine csgBAC and csgEFG into one synthetic operon. Without an RBS between csgBAC and csgEFG, translation of csgEFG would not occur. Of the infinite RBS's we could insert between csgBAC and csgEFG via Golden Gate assembly, we chose to use the sequence used in the successful curli circuit from "A Synthetic Circuit for Mercury Bioremediation Using Self-Assembling Functional Amyloids" (Tay, Nguyen, Joshi 2017).

This part was synthesized as an IDT Geneblock. When synthesized, BsaI cutsites and 4-basepair sticky ends were added to the ends of this part for 3G-Assembly compatibility. The part was further flanked by 40 basepair "Pad" adapter sequences that allow for insertion into William & Mary's "Pad" 1C3 backbone (via Gibson assembly) for cloning purposes. Once the sticky ends, cutsites, and Pad adapter sequences are added, this RBS is considered a composite part.

Source

This part was synthesized as an IDT Geneblock. The sequence was sourced from "A Synthetic Circuit for Mercury Bioremediation Using Self-Assembling Functional Amyloids" (Tay, Nguyen, Joshi 2017).

References

1. Barnhart, M. M., & Chapman, M. R. (2006). Curli biogenesis and function. Annual review of microbiology, 60, 131–147. doi:10.1146/annurev.micro.60.080805.142106

2. Tay, P. K. R., Nguyen, P. Q., & Joshi, N. S. (2017). A Synthetic Circuit for Mercury Bioremediation Using Self-Assembling Functional Amyloids. ACS Synthetic Biology, 6(10), 1841–1850. doi: 10.1021/acssynbio.7b00137