Plasmid

Part:BBa_K510014:Design

Designed by: David Caballero   Group: iGEM11_UPO-Sevilla   (2011-09-18)

pUC18R6KT-miniTn7BB-Km


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found in sequence at 4799
    Illegal suffix found in sequence at 1
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 4799
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal NotI site found at 9
    Illegal NotI site found at 4805
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 4799
    Illegal BglII site found at 3212
    Illegal BglII site found at 3483
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found in sequence at 4799
    Illegal suffix found in sequence at 2
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found in sequence at 4799
    Illegal XbaI site found at 4814
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1854
    Illegal SapI site found at 518


Design Notes

NcoI and SphI sites were added at the ends of the chloramphenicol resistance cassette to facilitate cloning

Source

This construct is the result of replacing the gentamycin resistance cassette in pUC18R6KT-miniTn7BB-Gm (BBa_K510012) by a kanamycin resistance cassette obtained from pBS4K5 by PCR amplification with these primers: tatGCATGCCCATGGattggggctcactcaaagg and tatGCATGCCCATGGtcgacaatgtaactactag.

References

Kyoung-Hee Choi, Jared B Gaynor, Kimberly G White, Carolina Lopez, Catharine M Bosio, RoxAnn R Karkhoff-Schweizer & Herbert P Schweizer (2005). A Tn7-based broad-range bacterial cloning and expression system. Nature Methods, vol.2 NO.6, June 2005, 443.