Composite

Part:BBa_J100307

Designed by: Monica Prudencio   Group: Campbell M Lab   (2016-10-08)


Variant of repClone Red (J100205)

This part is a variant of repClone Red (J100205), with some differences in the TetR and GFP.

We designed repClone Red to test the function of repressors. Internal BsaI sites allow the insertion of the Ptet promoter and removal of the backwards P2 promoter pointed towards GFP. Successful cloning of the Ptet promoter will change colonies from green to not green. Variations on the Ptet promoter, or introduction of anhydrotetrocycline (aTc), will affect the activity of the repressor, as indicated by the fluorescence of colonies containing the plasmid. This can be used in synthetic biology laboratories or for undergraduate introductory biology lab modules.

Variations from the registry sequence for TetR and GFP were sequence verified, and functionality was experimentally verified.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal SpeI site found at 9
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 948
    Illegal SpeI site found at 9
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal SpeI site found at 9
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal SpeI site found at 9
    Illegal AgeI site found at 2288
    Illegal AgeI site found at 2400
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1638
    Illegal BsaI.rc site found at 1517


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