Part:BBa_K510046:Design
pUC18R6KT-miniTn7BB-Gm-improved_flipflop_(module I)
- 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 NheI site found at 7687
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
Illegal BamHI site found at 5497 - 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
Illegal AgeI site found at 8949
Illegal AgeI site found at 9061 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 4828
Illegal BsaI.rc site found at 1854
Illegal SapI site found at 518
Design Notes
The BBa_K510019 BioBrick was inserted within the BCS of pUC18R6KT-miniTn7BB-Gm (K510012 BBa_K510012) by EcoRI and PstI clonning.
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. The improved flip-flop (module I) was synthesized commercially too.