Part:BBa_K1627000

Device to demethylate 1-methylxanthine

This part contains genes that should allow for the demethylation of 1-methylxanthine. The part contains a synthetic operon consisting of ndmA, ndmD from Pseudomonas putida CBB5 and gst9 from Janthinobacterium sp. Marseille under the control of a constitutive promoter, J23110.

While this device should be able to demethylate the 1-methyl group on methylxanthines, the ndmA gene lacks specificity to 1-methylxanthine. Thus, this part is not actually able to be used as designed. However, this part was important for the construction of parts BBa_K1627003, BBa_K1627004, and BBa_K1627006, which are able to demethylate the 1-methyl group from theophylline, paraxanthine, and caffeine, respectively.

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

Usage and Biology

This device was grown in the ∆guaB strain in minimal media cultures with different concentrations of 1-methylxanthine. No matter the concentration of 1-methylxanthine, the strain does not grow any substantial amount.

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
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 1199
  Illegal BglII site found at 1670
  Illegal XhoI site found at 2811
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 3509
  Illegal AgeI site found at 1338
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 983