Part:BBa_K3040112

pFadBA-Lac-2

We create a hybrid Fatty acid/acyl-CoA-regulated promoter by the combination of pLac with a modified promoter pFadBA. It is pointed out by the previous iGEM team, UPF CRG Barcelona_2018 iGEM, that the native promoter pFadBA has great leakage and is a weak promoter that isn’t ideal enough for our system. Therefore, to enhance the sensitivity and reduce the leakage of the promoter pFadBA for making it more suitable for our system, we improved our promoter based on the previous pfadBA DNA sequence from NTU_Taida 2012. pFL2 promoter has two fadR recognition sites, one between its -10 and -35 region, and the other lies just before the -35 region, with a promoter pLac behind -10 region. pFL2 promoter is designed to respond to changes of both fatty acid and an exogenous inducer, IPTG. The hybrid promoter pFL2 is fully activated only when both the fatty acids and IPTG are present.

Reference

Design of a dynamic sensor-regulator system for production of chemicals and fuels derived from fatty acids

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal prefix found in sequence at 30
  Illegal suffix found in sequence at 129
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 30
  Illegal SpeI site found at 130
  Illegal PstI site found at 144
  Illegal NotI site found at 36
  Illegal NotI site found at 137
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 30
  Illegal BglII site found at 50
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal prefix found in sequence at 30
  Illegal suffix found in sequence at 130
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal prefix found in sequence at 30
  Illegal XbaI site found at 45
  Illegal SpeI site found at 130
  Illegal PstI site found at 144
  
• 1000
  COMPATIBLE WITH RFC[1000]