Part:BBa_K801074

CaMXMT1 expression cassette for yeast

Yeast expression cassette for CaMXMT1 (BBa_K801071) controlled by the yeast TEF1 promoter (BBa_K319003) and the yeast ADH1 terminator (BBa_K801012).

This BioBrick is the generator for the enzyme 7-methylxanthine N-methyltransferase 1(CaMXMT1) (= theobromine synthase) and part of the caffeine synthesis pathway. It is regulated by the constitutive promoter Tef1, which is one of the strongest yeast promoters. The used terminator is Adh1, as it is among the other expression cassettes. The choice of the strong Tef1 promoter was made, because the in vivo biosynthesis of caffeine is an irreversible reaction, which is ensured by continously increasing Km and vmax values of the single enzymes. As soon as a certain amount of theobromine is available, the caffeine synthesis can go on. To support this irreversible, stepwise caffeine synthesis, we used the strongest promoter for this enzyme, to establish high concentrations of the caffeine precursor theobromine.

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 1433
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 205