Composite

Part:BBa_K4345024:Design

Designed by: Luka Van den Berghe   Group: iGEM22_KU_Leuven   (2022-10-08)
Revision as of 18:04, 8 October 2022 by LVdB (Talk | contribs)

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5' UTR of prfA with ccdA fused to sfGFP


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1229
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 649
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Our team used the RNA thermometer as a part of a killswitch. The sequence was implemented before an antitoxin (ccdB) to regulate its translation based on temperature. It is possible to implement a part of the sequence of the gene that will be translated within the sequence that forms secondary hairpin structure. By doing this, leaky expression will be reduced. Because of the typical short sequence of this thermometer and the position of the startcodon, our team decided not to. The position of the startcodon can be found in the provided figure.


Source

L. monocytogenes

References

Johansson, J., Mandin, P., Renzoni, A., Chiaruttini, C., Springer, M., & Cossart, P. (2002, September). An RNA Thermosensor Controls Expression of Virulence Genes in Listeria monocytogenes. Cell, 110(5), 551–561. https://doi.org/10.1016/s0092-8674(02)00905-4