Plasmid

Part:BBa_K510000:Design

Designed by: David Caballero, Fernando Govantes   Group: iGEM11_UPO-Sevilla   (2011-09-16)
Revision as of 12:26, 17 September 2011 by Fgovrom (Talk | contribs)

pUC18Sfi-miniTn7BB-Gm


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found in sequence at 4367
    Illegal suffix found in sequence at 1
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 4367
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal NotI site found at 9
    Illegal NotI site found at 4373
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 4367
    Illegal BglII site found at 3051
    Illegal BglII site found at 3322
    Illegal BglII site found at 3608
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found in sequence at 4367
    Illegal suffix found in sequence at 2
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found in sequence at 4367
    Illegal XbaI site found at 4382
    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 inserted at the ends of the kanamycin resistance cassette to facilitate cloning


Source

The miniTn7BB-Gm minitransposon was synthesized commercially, and then digested at the flanking SfiI sites and cloned into SfiI-digested pUC18-Sfi. This results in the elimination of the complete pUC18-SfiI multi-cloning site, except for the duplicated SfiI sites that remain on both sides of the transposon, thus facilitating its transfer to other vectors.

References