Composite

Part:BBa_K5377800:Design

Designed by: Jiyue Hu   Group: iGEM24_ZJUT-China   (2024-09-21)


RF self-induced production system


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 1892
    Illegal PstI site found at 3143
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
    Illegal PstI site found at 1892
    Illegal PstI site found at 3143
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 4220
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 1892
    Illegal PstI site found at 3143
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 1892
    Illegal PstI site found at 3143
    Illegal AgeI site found at 1739
    Illegal AgeI site found at 3563
    Illegal AgeI site found at 3973
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

1. The ribB FMN riboswitch is from the Escherichia coli. If you want to use it in other strains, please ensure that the riboswitch RBS sequence can be recognized.
2. The titer of RF has only been validated in Escherichia coli.

Source

ribB FMN riboswitch is from E. coli.
ribA, ribB, ribD, ribE, ribC are from E. coli.
The pHG101 plasmid (contain J23106 promoter) is synthesized by RuiBiotech Co., Ltd.

References

[1]Serganov, A., Huang, L., & Patel, D. J. (2009). Coenzyme recognition and gene regulation by a flavin mononucleotide riboswitch. Nature, 458(7235), 233–237. https://doi.org/10.1038/nature07642
[2]Winkler, W. C., Cohen-Chalamish, S., & Breaker, R. R. (2002). An mRNA structure that controls gene expression by binding FMN. Proceedings of the National Academy of Sciences of the United States of America, 99(25), 15908-15913. https://doi.org/10.1073/pnas.212628899
[3] de Boer, H. A., Comstock, L. J., & Vasser, M. (1983). The tac promoter: a functional hybrid derived from the trp and lac promoters. Proceedings of the National Academy of Sciences of the United States of America, 80(1), 21–25. https://doi.org/10.1073/pnas.80.1.21
[4] JACOB, F., & MONOD, J. (1961). Genetic regulatory mechanisms in the synthesis of proteins. Journal of molecular biology, 3, 318–356. https://doi.org/10.1016/s0022-2836(61)80072-7
[5] Müller-Hill, B. (1996). The lac Operon: A Short History of a Genetic Paradigm. Berlin, New York: De Gruyter. https://doi.org/10.1515/9783110879476