Composite

Part:BBa_K4217003:Design

Designed by: Madeline Gruenberg   Group: iGEM22_WrightState   (2022-09-26)


mntP-Riboswitch-sfGFP


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 255
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 760
    Illegal SapI.rc site found at 898


Design Notes

The 5′-untranslated region (5′-UTR) of the mntP gene contains a conserved riboswitch element (Barrick et. al., 2004). Two versions of the promoter were characterized in Dambach et. al. 2015. The first, a shorter ~264bp version displayed higher basal expression and a modest, ~1.3-fold induction in response to manganese. The longer, ~660bp version had lower basal expression and a more robust, ~5.1-fold induction in response to manganese, and was thus selected as the manganese responsive element for this part. The pmntR promoter-response to manganese was further characterized by Waters et. al. 2011.

Source

All the individual parts have sources on the parts page, the construct itself was assembled by us with the expertise of our mentors. This part was synthesized as a geneblock by IDT.

References

Barrick, J. E., Corbino, K. A., Winkler, W. C., Nahvi, A., Mandal, M., Collins, J., Lee, M., Roth, A., Sudarsan, N., Jona, I., Wickiser, J. K., & Breaker, R. R. (2004). New RNA motifs suggest an expanded scope for riboswitches in bacterial genetic control. Proceedings of the National Academy of Sciences of the United States of America, 101(17), 6421–6426. https://doi.org/10.1073/pnas.0308014101

Dambach, M., Sandoval, M., Updegrove, T. B., Anantharaman, V., Aravind, L., Waters, L. S., & Storz, G. (2015). The ubiquitous yybP-ykoY riboswitch is a manganese-responsive regulatory element. Molecular cell, 57(6), 1099–1109. https://doi.org/10.1016/j.molcel.2015.01.035

Waters, L. S., Sandoval, M., & Storz, G. (2011). The Escherichia coli MntR miniregulon includes genes encoding a small protein and an efflux pump required for manganese homeostasis. Journal of bacteriology, 193(21), 5887–5897. https://doi.org/10.1128/JB.05872-11