Part:BBa_K845000

iGEM 2012 Grenoble Team proposes a new design and application to the pBAD promoter(paraBAD).

In order to create a biological AND gate the 2012 Grenoble team worked on the cAMP-CRP complex and arabinose inductible promoter.

The activity of the promoter was tested under several conditions ( http://2012.igem.org/Team:Grenoble/Biology/Protocols/AND_test ) using a GFP as a reporter (on a pUA66). We tested this activity in BW25113 ΔcyaA in different culture media (Glucose + Acetate; Glycerol; Acetate).

Usage and Biology

paraBAD is a promoter which can be activated by 2 molecules : CRP-cAMP complex and the AraC protein. It has two states; in absence of L-arabinose the paraBAD promoter is repressed by AraC, whereas the paraC promoter is activated (unless an excess of AraC is present). In presence of L-arabinose and the CRP-cAMP complex, the promoter is activated thus enabling the transcription of the downstream elements. This AND gate provides a filter to biological noise.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal XbaI site found at 500
  
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 344
  Illegal XhoI site found at 494
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal XbaI site found at 500
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal XbaI site found at 500
  Illegal AgeI site found at 179
  Illegal AgeI site found at 484
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 1169
  Illegal SapI site found at 161