Composite

Part:BBa_K389014:Design

Designed by: Jonas Aretz   Group: iGEM10_Bielefeld-Germany   (2010-09-28)
Revision as of 15:14, 16 October 2010 by Jaretz (Talk | contribs)

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VirA/G screening system test


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
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 1768


Design Notes

  • The virA gene is under the control of 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 kanamycin resistance by the constitutive promoter low
  • only for testing the screening principle of mutated virA screening

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.