Plasmid
Part:BBa_K510013:Design
Designed by: David Caballero Group: iGEM11_UPO-Sevilla (2011-09-18)
pUC18R6KT-miniTn7BB-Cm
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal prefix found in sequence at 4640
Illegal suffix found in sequence at 1 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 4640
Illegal SpeI site found at 2
Illegal PstI site found at 16
Illegal NotI site found at 9
Illegal NotI site found at 4646 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 4640
Illegal BglII site found at 3212
Illegal BglII site found at 3483
Illegal XhoI site found at 3569 - 23INCOMPATIBLE WITH RFC[23]Illegal prefix found in sequence at 4640
Illegal suffix found in sequence at 2 - 25INCOMPATIBLE WITH RFC[25]Illegal prefix found in sequence at 4640
Illegal XbaI site found at 4655
Illegal SpeI site found at 2
Illegal PstI site found at 16 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1854
Illegal SapI site found at 518
Design Notes
Source
The pUC18R6KT vector was amplified by PCR using as template a pTNS2 plasmid (GenBank accession number: AY884833) provided by Herbert P. Schweizer. The primers were designed to flanked the pUC18R6KT with SfiI restriction sites and allow the insertion of the mini-Tn7-Gm. Primers: attcGGCCTAGGCGGCCgtcgttttacaacgtcgtgac and attcGGCCGCCTAGGCCggaagcataaagtgtaaagcctg. The miniTn7BB-Gm minitransposon was synthesized commercially, and then digested at the flanking SfiI sites and cloned into SfiI-digested pUC18R6KT PCR products. In order to change the resistance cassettes NcoI digestions were made.