Composite

Part:BBa_K510043:Design

Designed by: David Caballero   Group: iGEM11_UPO-Sevilla   (2011-10-21)

pUC18Sfi-miniTn7BB-Gm-bistable_switch


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
    Illegal BglII site found at 7454
  • 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
    Illegal AgeI site found at 8041
    Illegal AgeI site found at 8153
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1681
    Illegal BsaI.rc site found at 6010
    Illegal SapI.rc site found at 2763


Design Notes

The BBa_K177038 BioBrick was inserted within the BCS of pUC18Sfi-miniTn7BB-Gm (BBa K510000) by EcoRI and PstI clonning. In order to construct the pUC18Sfi-miniTn7BB-Gm delivery plasmid, the miniTn7BB-Gm transposon was cleaved from the commercial plasmid pMA (Mr. Gene) with SfiI and ligated to SfiI-digested pUC18Sfi. This strategy removes all multi-cloning restriction sites from pUC18Sfi, except for the flanking SfiI, thus guaranteeing that the unique sites in the transposon are not duplicated. The remove of the multi-cloning restriction sites was verified by analytic digestions.

Source

The miniTn7BB-Gm minitransposon was synthesized commercially and pUC18SfiI is a commercial vector.

References