Part:BBa_K2442101:Design

Minimal pBAD promoter

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 241
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 76
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI site found at 58
  

Design Notes

In absence of L-arabinose the AraC dimer binds to pBAD operator half-sites O₂ and I₁ and represses transcription by excluding RNA polymerase from binding to pBAD or PC. Binding of L-arabinose causes a conformational change in the protein such that the DNA-binding domains of the dimer bind to adjacent I_{1</sub and I2 half-sites, resulting in transcription activation of downstream genes (Fig. 1).}

Figure 1: Regulation of the L-arabinose operon by arabinose. In the absence of L-arabinose, AraC dimer binds to O₂ and I₁ half sites causing DNA looping, which prevents RNA polymerase from accessing P_BAD. Upon binding of L–arabinose, the AraC dimer binds I₁ and I₂ half sites instead, allowing transcription of the polycistronic araBAD mRNA. araC is transcribed in opposite direction from araBAD, and is under control of the PC promoter. RNA polymerase and AraC compete for binding at O₁ and P_C. Adapted from Shleif (2000)

Source

PCR amplification from BBa_I0500.

===References===