Part:BBa_K389010

VirA receptor (expressed)

This BioBrick contains a VirA receptor from Agrobacterium tumefaciens C58 under the control of a medium strong constitutive promoter.

Usage and Biology

We used this BioBrick to mutate virA by error-prone PCR so the mutated VirA receptor is expressed and ready to be screened for several substances.

The screening system for a mutated virA gene contains this BioBrick in a plasmid with ColE1 ori and the BioBrick BBa_K389011 in a plasmid with R6K ori. Both plasmids will be transformed to and screened in E. coli EC100D. Plasmids with R6K ori are only replicated in pir+ or pir116 E. coli strains. Once a receptor with high sensitivity for a screened substance is found, the plasmids are isolated and transformed to e.g. E. coli TOP10. Because the R6K ori does not work in this strain because it is not a pir+ / pir116 strain, it is easy to separate the mutated virA BioBrick from the screening plasmid.

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  Illegal NheI site found at 647
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 1632
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 1768