Part:BBa_K801077

Caffeine Synthesis Pathway pTEF2-XMT1-tADH1-pTEF1-MXMT1-tADH1-pTEF2-DXMT1-tADH1

This BioBrick is the final "caffeine synthesis device". It contains all three necessary enzymes: CaXMT1, CaMXMT1 and CaDXMT1, i.e. it is made up of the three single expression cassettes for each enzyme (see also BBa_K801073, BBa_K801074 and [https://parts.igem.org/Part:BBa_K801075 BBa_K801075). It can be directly transformed into competent yeast cells or cloned in an adequate yeast genome integration vector.

The expression of the enzymes CaXMT1 (Coffea arabica xanthosine-N-methyl-transferase 1, BBa_K801070) and CaDXMT1 (Coffea arabica dimethyl-xanthine N-methyl transferase 1, = caffeine synthase, BBa_K801072) is controlled by the TEF2-promoter (BBa_K801010) and the ADH1-terminator (BBa_K801012)

The expression of CaMXMT1 (Coffea arabica methyl-xanthine N-methyl transferase 1, BBa_K801071) is controlled by the TEF1 promoter (BBa_K319003) and the ADH1-terminator (BBa_K801012)

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 3613
  Illegal BglII site found at 5829
  Illegal BglII site found at 5925
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 138
  Illegal NgoMIV site found at 4317
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 2385