Part:BBa_K1627002

Device to demethylate 7-methylxanthine

This part contains genes that allow for the demethylation of 7-methylxanthine. The part contains a synthetic operon consisting of ndmC, 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 7-methylxanthine in a solution. By growing the ∆guaB strain with this device in a minimal media with different known concentrations of 7-methylxanthine, a growth curve can be made that correlates culture growth with concentration of 7-methylxanthine. Then if the strain is grown in a minimal media with an unknown concentration of 7-methylxanthine, the growth can be compared to the growth curve to calculate the initial concentration of 7-methylxanthine in the media.

Usage and Biology

This device was grown in the ∆guaB strain in minimal media cultures with different concentrations of theophylline. As the concentration of theophylline increases (up to the point of saturation, after which the strain won't grow further), the culture density increases linearly. By comparing growth from unknown concentrations of theophylline to the standard curve, the concentration of theophylline can be measured.

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 498
  Illegal BglII site found at 994
  Illegal BglII site found at 1465
  Illegal XhoI site found at 2606
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 3304
  Illegal AgeI site found at 1133
  
• 1000
  COMPATIBLE WITH RFC[1000]