Plasmid

Part:BBa_K510002:Design

Designed by: Fernando Govantes   Group: iGEM11_UPO-Sevilla   (2011-09-16)
Revision as of 08:07, 22 September 2011 by Dcabpra (Talk | contribs) (→‎Design Notes)

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pUC18SfiI-miniTn7BB-Km


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found in sequence at 4638
    Illegal suffix found in sequence at 1
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 4638
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal NotI site found at 9
    Illegal NotI site found at 4644
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 4638
    Illegal BglII site found at 3051
    Illegal BglII site found at 3322
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found in sequence at 4638
    Illegal suffix found in sequence at 2
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found in sequence at 4638
    Illegal XbaI site found at 4653
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1681
    Illegal SapI.rc site found at 2763


Design Notes

NcoI and SphI sites were added at the ends of the kanamycin resistance cassette to facilitate cloning For more information: [http://2011.igem.org/Team:UPO-Sevilla/Foundational_Advances/MiniTn7/Experimental_Results/miniTn7BB_derivatives Construction of additional miniTn7BB derivatives].

Source

The pUC18Sfi-miniTn7BB-Km was created by replacing the gentamycin resistance cassette of pUC18Sfi-miniTn7-Gm (K510000) by a kanamycin resistance cassette amplified from pSB4K5 with the following primers: tatGCATGCCCATGGattggggctcactcaaagg and tatGCATGCCCATGGtcgacaatgtaactactag

References