Difference between revisions of "Part:BBa K801020:Design"
Line 18: Line 18:
 
<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.-->
−
ethanol inducible, KlADH4
+
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:'''
−
<!--*Other versions:[https://parts.igem.org/wiki/index.php?title=Part:BBa_?????? BBa_??????: Name_of_part] -->
+
* Other versions:[https://parts.igem.org/wiki/index.php?title=Part:BBa_K678001 BBa_K678001: PalcA, inducible ''Aspergillus nidulans'' promoter]
 
<!--*Related BioBricks:[https://parts.igem.org/wiki/index.php?title=Part:BBa_?????? BBa_??????: Name_of_part]
 
<!--*Related BioBricks:[https://parts.igem.org/wiki/index.php?title=Part:BBa_?????? BBa_??????: Name_of_part]
  
 
'''Cloning details:'''<br>
 
'''Cloning details:'''<br>
−
*Designed in RFC25
+
* Designed in RFC25
−
<!--*Mutation C889G to delete XbaI restriction site-->
+
* 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-->
+
*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
Line 41: Line 40:
 
'''Backbone:'''<br>
 
'''Backbone:'''<br>
 
*Backbone name: pSB1C3'
 
*Backbone name: pSB1C3'
−
<!--*Resistance: Amp/Cp/Kan/Tet-->
+
*Resistance: Cam
−
<!--*Copynumber: low/medium/high-->
+
*Copynumber: high
  
 
'''Protein coding:'''<br>
 
'''Protein coding:'''<br>
−
<!--*Protein: ?Name_of_gene_product? [Nucleotide 1 to ???]-->
+
-
 
<!--*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 ???.-->
Line 51: Line 50:
  
 
'''Enzymatic activity:'''
 
'''Enzymatic activity:'''
 +
-
 
<!--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-->
+
* Known and anticipated sefety issues: none
−
<!--Known and anticipated security issues: none/other.-->
+
* Known and anticipated security issues: none
  
 
'''Intellectual property:'''
 
'''Intellectual property:'''
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'''Source:'''<br>
 
'''Source:'''<br>
−
<!--*Commercial system: plasmid name, system name, company name-->
+
* Cloned from genomic DNA of ''Kluyveromyces lactis'' strain WSYC115
−
<!--*Plasmid: p???, provided by ?name_of_person?, ?institute/university?, ?country?-->
+
* '''Forward Primer:'''<br><code>5'- AGT CAG TCT AGA ACG ACG GTA CAT AAG AAT GAG G - 3'</code><br>
−
<!--*Preexisting BioBrick ?Bba_number?-->
+
<!--'''Reverse Primer:'''<br><code>5'- GGC TAA GGT ACC CAT ATG GTG TGT TGT ATG TTG
−
<!--*cDNA Clone: ?clone_name?, ?company_name?-->
+
TGT TGT TGG - 3'</code><br>-->
−
<!--*Synthesized by ?company_name?.-->
+
−
 
+
−
<!--'''Forward Primer:'''<br><code>5'- ??? - 3'</code><br>-->
+
−
<!--'''Reverse Primer:'''<br><code>5'- ??? - 3'</code><br>-->
+
  
 
'''Organism:'''<br>
 
'''Organism:'''<br>
−
<!--*Genesequence derived from ''?organism_name?''-->
+
*Genesequence 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?''-->

Revision as of 15:46, 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



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'

Organism:

  • Genesequence derived from Kluyveromyces lactis


References

Literature references:

Database references: