Part:BBa_K510012:Design
pUC18R6KT-miniTn7BB-Gm
- 10INCOMPATIBLE WITH RFC[10]Illegal prefix found in sequence at 4528
Illegal suffix found in sequence at 1 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 4528
Illegal SpeI site found at 2
Illegal PstI site found at 16
Illegal NotI site found at 9
Illegal NotI site found at 4534 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 4528
Illegal BglII site found at 3212
Illegal BglII site found at 3483
Illegal BglII site found at 3769 - 23INCOMPATIBLE WITH RFC[23]Illegal prefix found in sequence at 4528
Illegal suffix found in sequence at 2 - 25INCOMPATIBLE WITH RFC[25]Illegal prefix found in sequence at 4528
Illegal XbaI site found at 4543
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
More information about [http://2011.igem.org/Team:UPO-Sevilla/Foundational_Advances/MiniTn7/Experimental_Results/Design_and_Synthesis Design and Synthesis] and [http://2011.igem.org/Team:UPO-Sevilla/Foundational_Advances/MiniTn7/Experimental_Results/Constructions_of_Vectors Construction of delivery vectors for miniTn7BB] in these links.
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.
References
Kyoung-Hee Choi, Jared B Gaynor, Kimberly G White, Carolina Lopez, Catharine M Bosio, RoxAnn R Karkhoff-Schweizer & Herbert P Schweizer (2005). A Tn7-based broad-range bacterial cloning and expression system. Nature Methods, vol.2 NO.6, June 2005, 443.