Part:BBa_K2555003

pBAD-RBS-sfGFP

BBa_K2555002 contains pBAD (L-arabinose inducing promoter) with RBS and CueR ( a transcription factor that can bind on pcopA of BBa_K2555000). E. coli cells use CueR to regulate the cytoplasmic copper concentration. Arabinose concentration controls the expression of CueR in the construct. CueR behaves as a net activator or a net repressor under different copper concentrations(1). BBa_K2555002 and BBa_K2555000 are used for construction of BBa_K2555004. BBa_K2555003 contains PBAD promoter, RBS, and superfolder GFP. The construct is used to indirectly reflect the expression of cueR under different concentration of L-arabinose by measuring green fluorescent intensity with plate reader.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 1311
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 1250
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 1085
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI site found at 1067
  Illegal SapI.rc site found at 1355
  

References

(1)Danya J. Martell, Chandra P. Joshi, Ahmed Gaballa, Ace George Santiago, Tai-Yen Chen, Won Jung, John D. Helmann, and Peng Chen (2015) “Metalloregulator CueR biases RNA polymerase’s kinetic sampling of dead-end or open complex to repress or activate transcription” Proc Natl Acad Sci U S A. 2015 Nov 3; 112(44): 13467–13472.