Part:BBa_K4761236

report-U6-2(new)

Temperature-sensitive reporter unit U6-2 for pET28a vector. 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 U6-2(new)[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 XbaI site found at 4
  Illegal SpeI site found at 18
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal SpeI site found at 18
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 47
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal XbaI site found at 4
  Illegal SpeI site found at 18
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal XbaI site found at 4
  Illegal SpeI site found at 18
  
• 1000
  COMPATIBLE WITH RFC[1000]

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