Part:BBa_K801073

CaXMT1 expression cassette for yeast

Yeast expression cassette for CaXMT1 (BBa_K801070) controlled by the yeast TEF2 promoter (BBa_K801010) and the yeast ADH1 terminator (BBa_K801012).

This BioBrick is the generator for the enzyme xanthosine N-methyltransferase 1 (CaXMT1) and part of the caffeine synthesis pathway. For creating an expression cassette with all three enzymes of the caffeine synthesis pathway BBa_K801077 based on the substrate Xanthosine different promoters and terminators were assembled to each enzyme. CaXMT1 is regulated by the constitutive promoter Tef2, which is a strong yeast promoter. The used terminator Adh1, is a widely used yeast terminator. The Tef2 promoter was prefered compared to the Tef1 promoter (which is even stronger) in order to limit metabolic stress, which could result in a positive selection of natural mutants (with regard to genome integration).

Cloning into pSB1C3

The cloning into pSB1C3 was proved by performing an analytical digest.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

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