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Part:BBa_J100111

Designed by: Phoebe Parrish   Group: Campbell M Lab   (2013-06-28)
Revision as of 13:41, 28 June 2013 by Phparrish (Talk | contribs)

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eCDM8 and Tetracycline Resistance Riboswitch into pSB1A8

High promoter/RBS combination with eCDM8 (part J100101) plus promoter/riboswitch/tetA gene construct (J119140), ligated into pSB1A8 (J119043). This part was constructed using iPCR and PCR to amplify the desired sequences and add BsaI sites, and Golden Gate Assembly to ligate the sequences together. eCDM8 demethylates caffeine to produce theophylline, which in turn binds to the riboswitch, allowing the translation of tetA mRNA, producing a protein that gives cells with this plasmid tetracycline resistance. Therefore cell growth can be used as a reporter of theophylline production.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 3578
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 237
    Illegal BamHI site found at 3724
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 3750
    Illegal NgoMIV site found at 4118
    Illegal NgoMIV site found at 4278
    Illegal AgeI site found at 196
    Illegal AgeI site found at 857
    Illegal AgeI site found at 3372
  • 1000
    COMPATIBLE WITH RFC[1000]


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