Part:BBa_K1627006

Device to demethylate caffeine

This part contains genes that allow for the demethylation of caffeine. The part contains a synthetic operon consisting of ndmA, ndmB, ndmC, and ndmD from Pseudomonas putida CBB5 and gst9 from Janthinobacterium sp. Marseille under the control of a constitutive promoter, J23110.

This part can be used in a ∆guaB strain of E. coli to measure the amount of caffeine in a solution. By growing the ∆guaB strain with this device in a minimal media with different known concentrations of caffeine, a growth curve can be made that correlates culture growth with concentration of caffeine. Then if the strain is grown in a minimal media with an unknown concentration of caffeine, the growth can be compared to the growth curve to calculate the initial concentration of caffeine in the media.

This part is an improved version of BBa_734000 that has the same function. Further information on the changes made can be found on the page for BBa_K734000.

See the [http://2015.igem.org/Team:Austin_UTexas/Project/Caffeine 2015 Austin UTexas wiki].

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  Illegal NheI site found at 1733
  Illegal NheI site found at 1928
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 2684
  Illegal BglII site found at 3180
  Illegal BglII site found at 3651
  Illegal XhoI site found at 4792
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 5490
  Illegal AgeI site found at 3319
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 983