Coding

Part:BBa_K1129019

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

Complete caffeine synthesis pathway under pTET constitutive promoter

We combined the three caffeine synthesis genes in the Registry by standard assembly into a pSB1C3 vector. This vector had a constitutive promoter, BBa_J23118, and a bacterial consensus sequence cloned into it previously, so that the completed construct would allow constitutive expression of the entire caffeine biosynthesis pathway when the substrate xanthosine was present. The strep tags were not removed to allow for detection of the protein products.

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 7
    Illegal NheI site found at 30
    Illegal NheI site found at 1373
    Illegal NheI site found at 1396
    Illegal NheI site found at 2757
    Illegal NheI site found at 2780
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2405
    Illegal BglII site found at 3807
    Illegal BglII site found at 3903
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None