Intermediate

Part:BBa_K558000:Design

Designed by: Jovanna Casas   Group: iGEM11_Nevada   (2011-08-25)

PDC/ADH coding regions


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NotI site found at 1727
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 1735
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1031
    Illegal AgeI site found at 1178
    Illegal AgeI site found at 1556
    Illegal AgeI site found at 2327
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 65
    Illegal BsaI site found at 2678
    Illegal SapI site found at 479
    Illegal SapI site found at 1142


Design Notes

A stuffer region between the PDC and ADH coding regions includes restriction digest sites for NotI and XhoI.

Under a constitutive promoter (Sigma 70 iGEM part J23101), this part has been shown to produce aldehydes (pdc) and 0.02% ethanol (adh).

Source

The sequence for these genes is based on Zymomonas mobilis, a microorganism that naturally produces ethanol.

References

Conway T, Sewell G W, Osman Y A, Ingram L O. Cloning and sequencing of the alcohol dehydrogenase II gene from Zymomonas mobilis. J Bacteriol. 1987;169:2591–2597.

Ingram, L.O., Conway, T., Clark, D.P., Sewell, G.W. and Preston, J.F. (1987, Oct) Genetic Engineering of Ethanol Production in Escherichia Coli. Applied and Environmental Microbiology: 2420-2425.