Difference between revisions of "Part:BBa K510013:Design"

 
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__NOTOC__
 
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<partinfo>BBa_K510013 short</partinfo>
 
<partinfo>BBa_K510013 short</partinfo>
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===Design Notes===
 
===Design Notes===
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The pUC18R6KT-mini-Tn7-Cm is a derivative of pUC18R6KT-mini-Tn7-Gm (BBa_K510012)
 
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by replacing the gentamycin resistance cassette by a chloramphenicol resistance cassette amplified from pSB1C3 with these primers: tatGCATGCCCATGGaacttggtctgacagctcgag and tatGCATGCCCATGGtcgggcacgtaagaggttcc.
 
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===Source===
 
===Source===

Revision as of 19:07, 18 September 2011

pUC18R6KT-miniTn7BB-Cm


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found in sequence at 4640
    Illegal suffix found in sequence at 1
  • 12
    INCOMPATIBLE 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
  • 21
    INCOMPATIBLE 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
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found in sequence at 4640
    Illegal suffix found in sequence at 2
  • 25
    INCOMPATIBLE 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
  • 1000
    INCOMPATIBLE 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.

References