Part:BBa_K783067

pBad-pTetR inverter

This is an inverter with pBad driving tetR with GFP as a reporter. pTetR has RFP has a reporter. We used a weaker RBS (B0032) to control the tetR expression and it helped the inverter function properly.

We used flow cytometry to measure the function of our inverter. As arabinose concentration increases, tetR and GFP also increases. As the tetR amount increases, the RFP decreases as tetR represses the pTetR promoter controlling RFP.

This result showed an improvement over an inverter we built with the B0034 RBS controlling the tetR gene (shown below). The gradual change in GFP and RFP seen above shows a finer control over the circuit than the dramatic drop off of RFP below. The GFP below is also very erratic.

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 1247
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 1187
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 979
  Illegal AgeI site found at 3490
  Illegal AgeI site found at 3602
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 2634
  Illegal SapI site found at 961