Part:BBa_K814013
DXMT1 generator
The induction of this novel pathway in S. cerevisiae requires two additional enzymes from Coffea canephora: XMT1 and DXMT1. S. cerevisiae will produce the required metabolic precursors to Xanthosine, where after the XMT1 enzyme from C. canephora will use the substrate to synthesize 7-methylxanthosine. DXMT1 will continue the synthesis by converting to 7-Methylxanthine, where in XMT1 will again convert it to Theobromine. Finally DXMT1 will convert the metabolite to caffeine. Both the XMT1 and DXMT1 genes produce bifunctional enzymes and this particular pathway was chosen so that one less enzyme would be required in the synthesis (compared to three in C. arabica).
This part includes the yeast pCyc promoter, a yeast Kozak sequence, the Coffea canephora XMT1 open reading frame which has been codon-optimized for yeast, and the tCycE1 terminator.
Figure 1. DXMT generator cassette assembled with IDT gBlocks. The cassette consists of the ADH1 promoter, the DXMT ORF and the ADH1 temrinator.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NotI site found at 1873
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 688
Illegal BamHI site found at 694
Illegal XhoI site found at 1861 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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