Part:BBa_K814012
XMT1 protein 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. It can be used to generate XMTI protein in yeast.
Figure 1. XMT generator. The XMT cassette is shown as assembled by IDT gBLocks. The cassette consists of the CYC1 promoter, the XMT ORF and the CYC1 terminator.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NotI site found at 1400
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 251
Illegal BamHI site found at 257
Illegal XhoI site found at 1388 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
| None |