Part:BBa_K4761234

report-U9

Temperature-sensitive reporter unit U9. Activates at around 37℃.

We inserted the RNA thermometer element into the plasmid encoding the EGFP gene by enzyme cleavage and enzyme link, and the specific position was between the promoter and the start codon to obtain the combined part REPORT U9[1][2]. Through the construction of this combination part, we can use the EGFP gene expression strength to characterize the success of the RNA thermometer. The strains transferred to the plasmid were cultured at different temperatures, and the expression intensity of EGFP was obtained at different temperatures, which reflected the opening temperature of the RNA thermometer.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 1
  Illegal SpeI site found at 20
  Illegal PstI site found at 26
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 1
  Illegal SpeI site found at 20
  Illegal PstI site found at 26
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 1
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 1
  Illegal SpeI site found at 20
  Illegal PstI site found at 26
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 1
  Illegal SpeI site found at 20
  Illegal PstI site found at 26
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 12
  

References

[1]Neupert, J., Bock, R. Designing and using synthetic RNA thermometers for temperature-controlled gene expression in bacteria. Nat Protoc 4, 1262–1273 (2009). [2]Juliane Neupert, Daniel Karcher, Ralph Bock, Design of simple synthetic RNA thermometers for temperature-controlled gene expression in Escherichia coli, Nucleic Acids Research, Volume 36, Issue 19, 1 November 2008, Page e124, https://doi.org/10.1093/nar/gkn545