Part:BBa_K615003

E. coli strain for TolC one-hybrid selection system

This strain is designed to test zinc finger binding by tying the binding event to cell survival, specifically the expression of TolC, a membrane pump that removes toxins from the cell. The omega subunit of RNA polymerase has been knocked out using lambda red to replace the gene with a zeocin cassette, and the strain is designed to be used in conjunction with a zinc finger-omega subunit fusion protein. See below for a summary of features and techniques.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 632
  Illegal PstI site found at 952
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 632
  Illegal PstI site found at 952
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 845
  Illegal BglII site found at 965
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 632
  Illegal PstI site found at 952
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 632
  Illegal PstI site found at 952
  Illegal NgoMIV site found at 181
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI site found at 121
  Illegal SapI site found at 331
  

Functional Parameters