Part:BBa_K2725005

Triclosan Resistant Backbone with RFP (pSB1Tcs1)

FabV is an enoyl acyl-carrier-protein reductase of Vibrio species, which is involved in type 2 fatty acid synthesis. It is a target for the biocide triclosan, which suppresses growth of E. coli DH5-alpha on agar plates at concentrations above 1 μM. When heterologously expressed in E. coli, this confers resistance to as it binds to triclosan, so the enoyl ACP reductase of E. coli (FabI) is free to function. Using Gibson Assembly, we replaced the chloramphenicol acetyltransferase gene of pSB1C3 with the fabV gene of Vibrio fischeri, creating a new plasmid backbone which confers resistance to triclosan.

Sequence and Features

Assembly Compatibility:

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

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 2592
  Illegal SpeI site found at 2
  Illegal PstI site found at 16
  Illegal NotI site found at 9
  Illegal NotI site found at 2598
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 2592
  Illegal XhoI site found at 1033
  Illegal XhoI site found at 2468
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal prefix found at 2592
  Illegal suffix found at 2
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal prefix found at 2592
  Plasmid lacks a suffix.
  Illegal XbaI site found at 2607
  Illegal SpeI site found at 2
  Illegal PstI site found at 16
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.