Coding

Part:BBa_K1129020

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

Complete caffeine synthesis pathway under arabinose-induced promoter

We combined the three caffeine synthesis genes in the Registry by standard assembly into a pSB1C3 vector. This vector had an arabinose-inducible promoter and a bacterial consensus sequence cloned into it previously, so that the completed construct would allow expression of the entire caffeine biosynthesis pathway when the substrate xanthosine and inducer arabinose was present.

Biosynthesis

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
    Illegal NheI site found at 1586
    Illegal NheI site found at 3065
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2595
    Illegal BglII site found at 4092
    Illegal BglII site found at 4188
    Illegal BamHI site found at 65
    Illegal BamHI site found at 1526
    Illegal BamHI site found at 3005
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None