Coding

Part:BBa_K1129017

Designed by: UBC iGEM 2013   Group: iGEM13_British_Columbia   (2013-08-30)

CaDXMT1 under arabinose-induced promoter

CaDXMT1 is the gene coding for the third and last enzyme of the caffeine biosynthesis pathway, known as 3,7-dimethylxanthine N-methyltransferase or caffeine synthase, when starting with xanthosine as substrate. TU Munich's 2012 team isolated this gene and added a strep tag at the end, in addition to transferring it all into yeast. We modified it further by removing the yeast consensus sequence and placing the gene and strep tag behind an arabinose-inducible promoter and bacterial consensus sequence. This system was transformed into a prokaryote, specifically E. coli strain K12.

Biosynthesis

Modifications

Characterization


Figure 1. 12% SDS-PAGE gel with protein samples from extracted from E. cloni® 10G cells containing each of the caffeine constructs (under pBAD) induced with 2% arabinose for 3 hours at 37°C. No additional bands are visible in the expected range when compared to the uninduced control.
Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 125
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1152
    Illegal BglII site found at 1248
    Illegal BamHI site found at 65
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None