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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+
−
 
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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.
+
−
 
+
−
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
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<br>'''Keywords:'''
 
<br>'''Keywords:'''
 
<!--These keywords are necessary to find your part using a fulltext sarch.-->
 
<!--These keywords are necessary to find your part using a fulltext sarch.-->
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ethanol inducible, KlADH4
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ethanol inducible, KlADH4, yeast, promoter, inducible, ''K. lactis'', ''S. cerevisiae''
 
<br>'''Abbreviations:'''
 
<br>'''Abbreviations:'''
 
<!--*used_abbreviation_1 = full_name_of_used_abbreviations_1-->
 
<!--*used_abbreviation_1 = full_name_of_used_abbreviations_1-->
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'''Related BioBrick:'''
 
'''Related BioBrick:'''
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<!--*Other versions:[https://parts.igem.org/wiki/index.php?title=Part:BBa_?????? BBa_??????: Name_of_part] -->
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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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*Designed in RFC25
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* Designed in RFC25
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<!--*Mutation C889G to delete XbaI restriction site-->
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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
−
<!--*Truncation upstream/downstream compared to template, ?explanation?-->
+
  
 
'''Quality control measures:'''<br>
 
'''Quality control measures:'''<br>
−
<!--*Test digestion using ?enzyme1? & ?enzyme2?/Not yet performed-->
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*Test digestion using EcoRI and SpeI
 
<!--*Sequencing using primer ?primer_name?/Not yet sequenced-->
 
<!--*Sequencing using primer ?primer_name?/Not yet sequenced-->
 
*Part was totally sequenced
 
*Part was totally sequenced
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'''Backbone:'''<br>
 
'''Backbone:'''<br>
 
*Backbone name: pSB1C3'
 
*Backbone name: pSB1C3'
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<!--*Resistance: Amp/Cp/Kan/Tet-->
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*Resistance: Cam
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<!--*Copynumber: low/medium/high-->
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*Copynumber: high
  
 
'''Protein coding:'''<br>
 
'''Protein coding:'''<br>
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<!--*Protein: ?Name_of_gene_product? [Nucleotide 1 to ???]-->
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-
 
<!--*The protein has the amino acid replacements ???99??? to ???99???.-->
 
<!--*The protein has the amino acid replacements ???99??? to ???99???.-->
 
<!--*The protein encoded is posttranslationally modified by ???.-->
 
<!--*The protein encoded is posttranslationally modified by ???.-->
 
<!--*Tag: n-terminally fused/c-terminally fused His5/His6/Strep/Flag/other-->
 
<!--*Tag: n-terminally fused/c-terminally fused His5/His6/Strep/Flag/other-->
  
−
'''Enzymatic activity:'''
+
'''Enzymatic activity:''' <br>
 +
-
 
<!--none/EC-number ?.?.?.?-->
 
<!--none/EC-number ?.?.?.?-->
  
 
'''Cytotoxicity:'''<br>
 
'''Cytotoxicity:'''<br>
 +
* none
 
<!--none/not known/cytotoxic for ''organism name''-->
 
<!--none/not known/cytotoxic for ''organism name''-->
  
 
'''Safety notes:'''<br>
 
'''Safety notes:'''<br>
−
<!--Known and anticipated sefety issues: none/health_risk/environmental_risk/other_risk-->
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* Known and anticipated sefety issues: none
−
<!--Known and anticipated security issues: none/other.-->
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* Known and anticipated security issues: none
  
 
'''Intellectual property:'''
 
'''Intellectual property:'''
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'''Source:'''<br>
 
'''Source:'''<br>
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<!--*Commercial system: plasmid name, system name, company name-->
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* Cloned from genomic DNA of ''Kluyveromyces lactis'' strain WSYC115
−
<!--*Plasmid: p???, provided by ?name_of_person?, ?institute/university?, ?country?-->
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* '''Forward Primer:'''<br><code>5'- AGT CAG TCT AGA ACG ACG GTA CAT AAG AAT GAG G - 3'</code><br>
−
<!--*Preexisting BioBrick ?Bba_number?-->
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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>
−
<!--*cDNA Clone: ?clone_name?, ?company_name?-->
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−
<!--*Synthesized by ?company_name?.-->
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−
 
+
−
<!--'''Forward Primer:'''<br><code>5'- ??? - 3'</code><br>-->
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−
<!--'''Reverse Primer:'''<br><code>5'- ??? - 3'</code><br>-->
+
  
 
'''Organism:'''<br>
 
'''Organism:'''<br>
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<!--*Genesequence derived from ''?organism_name?''-->
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* Sequence derived from ''Kluyveromyces lactis''
 
<!--*Codonoptimized for ''?organism_name?''-->
 
<!--*Codonoptimized for ''?organism_name?''-->
 
<!--*Designed for the following Chassis: ''?organism-name?''-->
 
<!--*Designed for the following Chassis: ''?organism-name?''-->
 
<!--*Statement about functionality in other chassis.-->
 
<!--*Statement about functionality in other chassis.-->
−
 
  
 
===References===
 
===References===
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'''Literature references:'''<br>
 
'''Literature references:'''<br>
−
<!--*[http://www.ncbi.nlm.nih.gov/pubmed/?PMID? '''Pubmed:''' ?Author(s)?, ?year?: ?title?]-->
+
* [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]
  
−
'''Database references:'''<br>
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* [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]
−
<!--*[http://www.ncbi.nlm.nih.gov/nuccore/?accessNr? '''GenBank''': ?title?]-->
+
−
<!--*[http://www.ebi.ac.uk/interpro/IEntry?ac=?accessNr? '''Interpro''': ?title?]-->
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−
<!--*[http://www.uniprot.org/uniprot/?accessNr? '''Uniprot''': ?title?]-->
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−
<!--*[http://pfam.sanger.ac.uk/family/?accessNr? '''Pfam:''' ?title?]-->
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<!--*[http://www.rcsb.org/pdb/explore/explore.do?structureId=?accessNR? '''PDB:''' ?tile?]-->
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<!--*[http://www.brenda-enzymes.info/php/result_flat.php4?ecno=?accessNr? '''Branda:''' ?title?]-->
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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]