RNA
Part:BBa_K4811029:Design
Designed by: Kasper Krunderup Jakobsen Group: iGEM23_DTU-Denmark (2023-10-03)
TMS(Mn2)
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 147
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 105
Design Notes
The RNA has to fold as a tRNA would.
Multiple versions of a Mn-sensitive TMS were attempted, in this instance, only the manganese binding element was inserted into the D-loop.
Source
The TMS was developed by Avishek Paul during their PhD thesis, see the publication "Modular and Versatile Trans-Encoded Genetic Switches", A. Paul, E. M. Warszawik, M. Loznik, A. J. Boersma, A. Herrmann, Angew. Chem. Int. Ed. 2020, 59, 20328.
The manganese aptamer was adapted from 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