Composite
Part:BBa_K510042:Design
Designed by: David Caballero, Fernando Govantes Group: iGEM11_UPO-Sevilla (2011-10-19)
pUC18Sfi-miniTn7BB-Gm-Lux
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal prefix found in sequence at 4367
Illegal suffix found in sequence at 1 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 4367
Illegal NheI site found at 5601
Illegal NheI site found at 8567
Illegal SpeI site found at 2
Illegal PstI site found at 16
Illegal NotI site found at 9
Illegal NotI site found at 4373 - 21INCOMPATIBLE 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 7565
Illegal BamHI site found at 5540
Illegal XhoI site found at 8395 - 23INCOMPATIBLE WITH RFC[23]Illegal prefix found in sequence at 4367
Illegal suffix found in sequence at 2 - 25INCOMPATIBLE 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 5375 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 1681
Illegal BsaI site found at 9954
Illegal BsaI.rc site found at 6963
Illegal SapI site found at 5357
Illegal SapI.rc site found at 2763
Illegal SapI.rc site found at 10279
Design Notes
The BBa_K325909 BioBrick was inserted within the BCS of pUC18Sfi-miniTn7BB-Gm 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.