Coding
THI3

Part:BBa_J45009:Design

Designed by: Veena Venkatachalam   Group: iGEM06_MIT   (2006-06-07)

ketoisocaproate decarboxylase (THI3); converts alpha-ketoisocaproate to 3-methylbutanal


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 3
    Illegal EcoRI site found at 904
    Illegal EcoRI site found at 1603
    Illegal EcoRI site found at 1789
    Illegal SpeI site found at 697
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 3
    Illegal EcoRI site found at 904
    Illegal EcoRI site found at 1603
    Illegal EcoRI site found at 1789
    Illegal SpeI site found at 697
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 3
    Illegal EcoRI site found at 904
    Illegal EcoRI site found at 1603
    Illegal EcoRI site found at 1789
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 3
    Illegal EcoRI site found at 904
    Illegal EcoRI site found at 1603
    Illegal EcoRI site found at 1789
    Illegal SpeI site found at 697
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 3
    Illegal EcoRI site found at 904
    Illegal EcoRI site found at 1603
    Illegal EcoRI site found at 1789
    Illegal SpeI site found at 697
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

  • See [http://pathway.yeastgenome.org:8555/YEAST/NEW-IMAGE?type=PATHWAY&object=PWY3O-4112 biosynthetic pathway from leucine to isoamyl alcohol] (the precursor to isoamyl acetate or banana odor)
  • Constructed by PCR

Template

Saccharomyces cerevisiae strain ACLY387 genome

Forward primer

5'- GTT TCT TCG AAT TCG CGG CCG CTT CTA Gat gaa ttc tag cta tac aca gag -3'

Reverse primer

5'- GTT TCT TCC TGC AGC GGC CGC TAC TAG TAT TAT TAg tat cca act tga ttt ttt ttt aga ag -3'

Source

  • THI3 from Saccharomyces cerevisiae (GenBank [http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?val=1431099&from=520&to=2349&view=gbwithparts Z74128 REGION: 520..2349])
  • [http://db.yeastgenome.org/cgi-bin/locus.pl?sgdid=S000002238 THI3 SGD entry]

References

Refers only indirectly to THI3 Dickinson-JBC-1997. <biblio>

  1. Dickinson-JBC-1997 pmid=9341119
  2. Yoshimoto-ApplMicrobiolBiotechnol-2002 pmid=12172617
  3. Nishimura-1992 pmid=1624458
  4. Lilly--2006 pmid=16879424

</biblio>