Plasmid

Part:BBa_K734005

Designed by: Ben Slater   Group: iGEM12_Austin_Texas   (2012-10-01)


pReporter for ZombiE.coli

This plasmid contains a lacUV5 promoter between two mutant, inverted loxP sites (lox66 and lox77). When Cre recombinase is present, recombination flips the region between the loxP sites, causing the promoter to be oriented the opposite direction. This recombination should only be capable of occuring once, because after recombination one loxP site becomes wild-type and the other becomes double-mutant, disallowing binding by Cre.

Before recombination, the promoter is oriented to express GFP, signaling healthy, "uninfected" cells. However, if pConverter expresses Cre (due to presence of the zombifying infectious agent AHL), the promoter is flipped to express RFP, signaling an "infected" cell. Downstream of rfp is lasI, which allows infected cells to have the ability to infect others.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 2353
    Illegal PstI site found at 3300
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 2353
    Illegal PstI site found at 3300
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 2394
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 2353
    Illegal PstI site found at 3300
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 2353
    Illegal PstI site found at 3300
    Illegal NgoMIV site found at 5477
    Illegal NgoMIV site found at 5637
    Illegal AgeI site found at 568
    Illegal AgeI site found at 846
    Illegal AgeI site found at 958
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 3474
    Illegal BsaI.rc site found at 70
    Illegal SapI.rc site found at 4556


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