Composite

Part:BBa_K389011:Design

Designed by: Jonas Aretz   Group: iGEM10_Bielefeld-Germany   (2010-08-10)

VirA screening device


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

  • medium strong constitutive promoter
  • 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 kanamycin resistance by the constitutive promoter low
  • Kanamycin resistance for screening with agar plates - the more the vir promoter is activated by VirG the more resistant the cells become to kanamycin.

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.