Composite

Part:BBa_K389015:Design

Designed by: Jonas Aretz   Group: iGEM10_Bielefeld-Germany   (2010-09-28)

VirA/G reporter device luc


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
    Illegal NheI site found at 2581
    Illegal NheI site found at 2604
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1632
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 3769
    Illegal NgoMIV site found at 5113
    Illegal NgoMIV site found at 5134
    Illegal AgeI site found at 4837
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 1768
    Illegal SapI.rc site found at 5019


Design Notes

  • The virA gene is under the control a medium strong constitutive promoter so there is enough VirA receptor expressed but not too much so the cell suffers from the expression (VirA is a membrane protein)
  • The virG gene is mutated so it works without an rpoA subunit from Agrobacterium tumefaciens (YC Jung et al., 2004)
  • double terminator (forward) to keep expression of luciferase gene by the constitutive promoter low
  • luciferase gene to show the activity of the vir promoter
  • this BioBrick is for measuring the behaviour of the natural VirA/G system

Source

References

YC Jung et al. (2004) Mutants of Agrobacterium tumefaciens virG Gene That Activate Transcription of vir Promoter in Escherichia coli, Current Microbiol 49:334-340.