Part:BBa_K615000

E. coli strain for His3-URA3 one-hybrid selection system

This strain is designed to test zinc finger binding using a one-hybrid metabolic selection system. The endogenous genes HisB and PyrF have been knocked out and replaced with the yeast versions His3 and URA3 under the control of a Zif268 binding site. The gene coding for the omega subunit of RNA polymerase (rpoZ) was knocked out by replacing it with a Zeocin cassette, and the strain is designed to be used in conjunction with a zinc finger-omega subunit expression construct.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 632
  Illegal PstI site found at 952
  Illegal PstI site found at 1762
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 632
  Illegal PstI site found at 952
  Illegal PstI site found at 1762
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 845
  Illegal BglII site found at 965
  Illegal BglII site found at 1478
  Illegal BglII site found at 1538
  Illegal BamHI site found at 1768
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 632
  Illegal PstI site found at 952
  Illegal PstI site found at 1762
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 632
  Illegal PstI site found at 952
  Illegal PstI site found at 1762
  Illegal NgoMIV site found at 181
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 2458
  Illegal SapI site found at 121
  Illegal SapI site found at 331
  Illegal SapI.rc site found at 2305
  

Functional Parameters