Translational_Unit

Part:BBa_K734000:Design

Designed by: Erik Quandt   Group: iGEM12_Austin_Texas   (2012-08-07)

Caffeine demethylation pathway


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1229
    Illegal XbaI site found at 1126
    Illegal XbaI site found at 2229
    Illegal XbaI site found at 3109
    Illegal PstI site found at 548
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1229
    Illegal NheI site found at 8
    Illegal NheI site found at 31
    Illegal NheI site found at 1730
    Illegal NheI site found at 1925
    Illegal PstI site found at 548
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1229
    Illegal BglII site found at 2687
    Illegal BglII site found at 3181
    Illegal BglII site found at 3652
    Illegal XhoI site found at 4793
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1229
    Illegal XbaI site found at 1126
    Illegal XbaI site found at 2229
    Illegal XbaI site found at 3109
    Illegal PstI site found at 548
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1229
    Illegal XbaI site found at 1126
    Illegal XbaI site found at 2229
    Illegal XbaI site found at 3109
    Illegal PstI site found at 548
    Illegal NgoMIV site found at 5491
    Illegal NgoMIV site found at 5604
    Illegal AgeI site found at 2224
    Illegal AgeI site found at 3320
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 984
    Illegal BsaI.rc site found at 2221


Design Notes

Genes and were amplified and assembled using Gibson isothermal assembly. BBa_K515105 was used as a source for the J23100 promoter + pSB1C3 backbone.

Source

Methylxanthine N-demethylases NdmA,NdmB,NdmC and associated reductase NdmD were amplified from Psuedomonas putida CBB5 genomic DNA.

gst9 from Janthinobacterium sp. Marseille was synthesized.

References

<biblio>

  1. Summers2012 pmid=22328667
  2. Summers2011 pmid=20966097
  3. Yu2009 pmid=19447909

</biblio>