Difference between revisions of "Part:BBa K801020:Design"

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<partinfo>BBa_K801020 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K801020 SequenceAndFeatures</partinfo>
  
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The feature annotation in the UASe region (Upstream activating sequence "ethanol") is taken from Mazzoni et al.,2000.
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To remove a naturally occuring SpeI-restriction site, the naturally occuring "A" at position 757 was changed to "T" using quickchange mutagenesis.
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The feature annotation in the UASe region (Upstream activating sequence "ethanol") is taken from:
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* 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
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'''Related BioBrick:'''
 
'''Related BioBrick:'''
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* Other versions:[https://parts.igem.org/wiki/index.php?title=Part:BBa_K678001 BBa_K678001: PalcA, inducible ''Aspergillus nidulans'' promoter]
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* Related biobrick: [https://parts.igem.org/wiki/index.php?title=Part:BBa_K678001 BBa_K678001]: PalcA, inducible ''Aspergillus nidulans'' promoter
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<!--*Related BioBricks:[https://parts.igem.org/wiki/index.php?title=Part:BBa_?????? BBa_??????: Name_of_part]
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<!--*Related BioBricks:[https://parts.igem.org/wiki/index.php?title=Part:BBa_?????? BBa_??????: Name_of_part]--->
  
 
'''Cloning details:'''<br>
 
'''Cloning details:'''<br>
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<!--*Tag: n-terminally fused/c-terminally fused His5/His6/Strep/Flag/other-->
 
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'''Enzymatic activity:'''
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'''Enzymatic activity:''' <br>
 
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* Cloned from genomic DNA of ''Kluyveromyces lactis'' strain WSYC115
 
* Cloned from genomic DNA of ''Kluyveromyces lactis'' strain WSYC115
 
* '''Forward Primer:'''<br><code>5'- AGT CAG TCT AGA ACG ACG GTA CAT AAG AAT GAG G - 3'</code><br>
 
* '''Forward Primer:'''<br><code>5'- AGT CAG TCT AGA ACG ACG GTA CAT AAG AAT GAG G - 3'</code><br>
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* '''Reverse Primer:'''<br><code>5'- GGC TAA GGT ACC CAT ATG GTG TGT TGT ATG TTG  
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* '''Reverse Primer:'''<br><code>5'- GGC TAA GGT ACC CAT ATG GTG TGT TGT ATG TTG TGT TGT TGG - 3'</code><br>
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TGT TGT TGG - 3'</code><br>
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'''Organism:'''<br>
 
'''Organism:'''<br>

Latest revision as of 15:51, 22 October 2012

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

The feature annotation in the UASe region (Upstream activating sequence "ethanol") is taken from Mazzoni et al.,2000.


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:

  • Related biobrick: BBa_K678001: PalcA, inducible Aspergillus nidulans promoter

Cloning details:

  • Designed in RFC25
  • To remove a naturally occuring SpeI-restriction site, the naturally occuring "A" at position 757 was changed to "T" using quickchange mutagenesis

Quality control measures:

  • Test digestion using EcoRI and SpeI
  • Part was totally sequenced

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]