Regulatory

Part:BBa_K801020:Design

Designed by: Simon Heinze   Group: iGEM12_TU_Munich   (2012-09-21)
Revision as of 15:49, 22 October 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



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


Extracted from genomic DNA of K. lactis using PCR.


Keywords: ethanol inducible, KlADH4, yeast, promoter, inducible, K. lactis, S. cerevisiae
Abbreviations:

Design Notes

Related BioBrick:

Backbone:

  • Backbone name: pSB1C3'
  • Resistance: Cam
  • Copynumber: high

Protein coding:
-

Enzymatic activity:
-

Cytotoxicity:

  • none

Safety notes:

  • Known and anticipated sefety issues: none
  • Known and anticipated security issues: none

Intellectual property:

Corresponding part author/authors:

Source

Source:

  • Cloned from genomic DNA of Kluyveromyces lactis strain WSYC115
  • Forward Primer:
    5'- AGT CAG TCT AGA ACG ACG GTA CAT AAG AAT GAG G - 3'
  • Reverse Primer:
    5'- GGC TAA GGT ACC CAT ATG GTG TGT TGT ATG TTG TGT TGT TGG - 3'

Organism:

  • Sequence derived from Kluyveromyces lactis

References

Literature references:

  • [1] Saliola, M., Mazzoni, C., Solimando, N., Crisà, A., Falcone, C. & Jung, G. (1999) ‚Use of the KlADH4 promoter for ethanol-dependent production of recombinant human serum albumin in Kluyveromyces lactis’, Appl Environ Microbiol. 65 (1), 53-60. [http://www.ncbi.nlm.nih.gov/pubmed?term=10724480 PMID: 9872759]
  • [2] 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. [http://www.ncbi.nlm.nih.gov/pubmed?term=10724480 PMID: 10724480]