Part:BBa_K3519011

araC-araBAD promoter-mRFP

This part contains the RFP downstream to the arabinose inducible promoter. This has been designed keeping in mind a tightly regulated 'kill switch'. Look at the part BBa_K3519012 for the kill switch. The RFP gene here is expressed in the addition of arabinose to the media. Excess glucose in the media represses the expression of RFP.

Usage and Biology

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Figure 1: The ‘native’ genetic circuit used for characterisation of the designed arabinose inducible promoter system. The araC is constitutively transcribed in the opposite direction.



Figure 2: The ‘active’ genetic circuit. The system here is switched ‘on’ in addition of arabinose to the media. The araC is constitutively transcribed in the opposite direction. The promoter is activated on recruitment of the cAMP-CAP complex and the arabinose-AraC complex to the regulatory regions in the promoter, leading to increase in the transcription of the mRFP gene. The designed arabinose inducible promoter system is characterised by quantifying mRFP using fluorescence measurements.



Figure 3: The ‘inactive’/repressed genetic circuit. The system here is switched ‘off’ in absence of arabinose in the media. The araC gene is constitutively transcribed in the opposite direction. The AraC dimer binds to the regulatory regions of the promoter and represses the transcription of mRFP, leading to very low fluorescence.

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 1043
  Illegal NheI site found at 1066
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 1388
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 1223
  Illegal AgeI site found at 2030
  Illegal AgeI site found at 2142
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI site found at 1205