Cell

Part:BBa_K615003:Design

Designed by: Naomi Genuth   Group: iGEM11_Harvard   (2011-09-26)

E. coli strain for TolC one-hybrid selection system


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 632
    Illegal PstI site found at 952
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 632
    Illegal PstI site found at 952
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 845
    Illegal BglII site found at 965
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 632
    Illegal PstI site found at 952
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 632
    Illegal PstI site found at 952
    Illegal NgoMIV site found at 181
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 121
    Illegal SapI site found at 331


Design Notes

rpoZ knocked out using lambda red

Kanamycin cassette-Zif268 binding site-weak promoter placed upstream of endogenous TolC

Source

Based off EcNR2 selection strain (see Wang et al, 2009)

References

Harris H. Wang, Farren J. Isaacs, Peter A. Carr, Zachary Z. Sun, George Xu, Craig R. Forest, George M. Church. Programming cells by multiplex genome engineering and accelerated evolution. (2009). Nature, 460(7257):894-8.