DNA

Part:BBa_J100115

Designed by: Phoebe Parrish   Group: Campbell M Lab   (2013-07-13)


eCDM8 and GFP Riboswitch into pSB1A8

High promoter/RBS combination with eCDM8 (part J119303) plus promoter/riboswitch/GFP construct (J100079), 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 GFP mRNA, producing green fluorescent protein as a reporter of theophylline production.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 237
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 196
    Illegal AgeI site found at 857
    Illegal AgeI site found at 3371
    Illegal AgeI site found at 3455
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 3443


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Categories
Parameters
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