Regulatory

Part:BBa_K801020:Design

Designed by: Simon Heinze   Group: iGEM12_TU_Munich   (2012-09-21)
Revision as of 14:32, 24 September 2012 by Simon Heinze (Talk | contribs) (→‎Design Notes)

KlADH4 yeast promoter, ethanol inducible


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 753
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 590


Design Notes

To remove a naturally occuring SpeI-restriction site, the naturally occuring "A" at position 757 was changed to "T" using quickchange mutagenesis.

The feature annotation in the UASe region (Upstream activating sequence "ethanol") is taken from:

  • Mazzoni, C., Santori, F., Saliola, M. & Falcone, C. (2000) ‚Molecular analysis of UASE, a cis element containing stress response elements responsible for ethanol induction of the KlADH4 gene of Kluyveromyces lactis’, Res. Microbiol. 151, 19-28. PMID: 10724480

Source

Extracted from genomic DNA of K. lactis using PCR.

References