Difference between revisions of "Part:BBa K801073:Design"

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Composite part of BBa_K801010, BBa_K801070, BBa_K801012
 
Composite part of BBa_K801010, BBa_K801070, BBa_K801012
  
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<br>'''Keywords:'''
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<br>'''Keywords:''' CaXMT1, TEF2, ADH1, caffeine,
 
<!--These keywords are necessary to find your part using a fulltext sarch.-->
 
<!--These keywords are necessary to find your part using a fulltext sarch.-->
 
<!--keyword_1, keyword_2, keyword_3, keyword_4, keyword_5-->
 
<!--keyword_1, keyword_2, keyword_3, keyword_4, keyword_5-->
  
 
<br>'''Abbreviations:'''
 
<br>'''Abbreviations:'''
−
<!--*used_abbreviation_1 = full_name_of_used_abbreviations_1-->
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*CaXMT1 = xanthosine methyltransferase
 
<!--*used_abbreviation_2 = full_name_of_used_abbreviations_2-->
 
<!--*used_abbreviation_2 = full_name_of_used_abbreviations_2-->
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* CaXMT1 = coffea arabica xanthosine N-methyltransferase 1
  
 
===Design Notes===
 
===Design Notes===
  
 
'''Related BioBrick:'''
 
'''Related BioBrick:'''
−
<!--*Other versions:[https://parts.igem.org/wiki/index.php?title=Part:BBa_?????? BBa_??????: Name_of_part] -->
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*Other versions:
−
<!--*Related BioBricks:[https://parts.igem.org/wiki/index.php?title=Part:BBa_?????? BBa_??????: Name_of_part]
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**[https://parts.igem.org/Part:BBa_K801070 BBa_K801070: xanthosine methyltransferase CaXMT1-strep]
 +
*Related BioBricks:
 +
** [https://parts.igem.org/Part:BBa_K801071 BBa_K801071: 7-methylxanthine N-methyltransferase CaMXMT1-strep]
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** [https://parts.igem.org/Part:BBa_K801072 BBa_K801072: 3,7-dimethylxanthine N-methyltransferase CaDXMT1-strep]
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** [https://parts.igem.org/Part:BBa_K801073 BBa_K801073: CaXMT1 expression cassette for yeast]
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** [https://parts.igem.org/Part:BBa_K801074 BBa_K801074: CaMXMT1 expression cassette for yeast]
 +
** [https://parts.igem.org/Part:BBa_K801075 BBa_K801075: CaDXMT1 expression cassette for yeast]
 +
** [https://parts.igem.org/Part:BBa_K801076 BBa_K801076: Yeast expression cassette for CaXMT1 and CaMXMT1]
 +
** [https://parts.igem.org/Part:BBa_K801077 BBa_K801077: Caffeine Synthesis Pathway pTEF2-XMT1-tADH1-pTEF1-MXMT1-tADH1-pTEF2-DXMT1-tADH1]
  
 
'''Cloning details:'''<br>
 
'''Cloning details:'''<br>
−
<!--*Designed in RFC10/RFC23/RFC25/RFC25_N-part-->
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* Designed in RFC10
 
<!--*Mutation C889G to delete XbaI restriction site-->
 
<!--*Mutation C889G to delete XbaI restriction site-->
 
<!--*Truncation upstream/downstream compared to template, ?explanation?-->
 
<!--*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 XbaI & PstI
 
<!--*Sequencing using primer ?primer_name?/Not yet sequenced-->
 
<!--*Sequencing using primer ?primer_name?/Not yet sequenced-->
−
<!--*Part was partly sequenced/Part was totally sequenced-->
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*Part was totally sequenced
  
 
'''Backbone:'''<br>
 
'''Backbone:'''<br>
−
<!--*Backbone name: pSB1C3'/?backbone_name?-->
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* Backbone name: pSB1C3
−
<!--*Resistance: Amp/Cp/Kan/Tet-->
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* Resistance: Cp
−
<!--*Copynumber: low/medium/high-->
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* Copynumber: high
  
 
'''Protein coding:'''<br>
 
'''Protein coding:'''<br>
−
<!--*Protein: ?Name_of_gene_product? [Nucleotide 1 to ???]-->
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* Protein: CaXMT1 [Nucleotide 663 to 1778]
 
<!--*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-->
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* Tag: c-terminally fused Strep
  
 
'''Enzymatic activity:'''
 
'''Enzymatic activity:'''
−
<!--none/EC-number ?.?.?.?-->
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* EC-number: 2.1.1.158
  
 
'''Cytotoxicity:'''<br>
 
'''Cytotoxicity:'''<br>
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'''Source:'''<br>
 
'''Source:'''<br>
 +
* *Preexisting BioBrick BBa_K801070
 +
 
<!--*Commercial system: plasmid name, system name, company name-->
 
<!--*Commercial system: plasmid name, system name, company name-->
 
<!--*Plasmid: p???, provided by ?name_of_person?, ?institute/university?, ?country?-->
 
<!--*Plasmid: p???, provided by ?name_of_person?, ?institute/university?, ?country?-->
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'''Organism:'''<br>
 
'''Organism:'''<br>
−
<!--*Genesequence derived from ''?organism_name?''-->
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* Genesequence derived from ''coffea arabica''
−
<!--*Codonoptimized for ''?organism_name?''-->
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* Codonoptimized for ''Saccharomyces cerevisiae''
−
<!--*Designed for the following Chassis: ''?organism-name?''-->
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* Designed for the following Chassis: ''Saccharomyces cerevisiae''
 
<!--*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?]-->
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* Uefuji, H., Ogita, S., Yamaguchi, Y., Koizumi, N., and Sano, H. (2003). Molecular cloning and functional characterization of three distinct n-methyltransferases involved in the caffeine biosynthetic pathway in coffee plants. Plant Physiol, 132(1):372–80.
  
 
'''Database references:'''<br>
 
'''Database references:'''<br>
 +
* NCBI
 
<!--*[http://www.ncbi.nlm.nih.gov/nuccore/?accessNr? '''GenBank''': ?title?]-->
 
<!--*[http://www.ncbi.nlm.nih.gov/nuccore/?accessNr? '''GenBank''': ?title?]-->
 
<!--*[http://www.ebi.ac.uk/interpro/IEntry?ac=?accessNr? '''Interpro''': ?title?]-->
 
<!--*[http://www.ebi.ac.uk/interpro/IEntry?ac=?accessNr? '''Interpro''': ?title?]-->

Latest revision as of 10:36, 24 October 2012

CaXMT1 expression cassette for yeast


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 138
  • 1000
    COMPATIBLE WITH RFC[1000]


Composite part of BBa_K801010, BBa_K801070, BBa_K801012


Keywords: CaXMT1, TEF2, ADH1, caffeine,


Abbreviations:

  • CaXMT1 = xanthosine methyltransferase
  • CaXMT1 = coffea arabica xanthosine N-methyltransferase 1

Design Notes

Related BioBrick:

Cloning details:

  • Designed in RFC10

Quality control measures:

  • Test digestion using XbaI & PstI
  • Part was totally sequenced

Backbone:

  • Backbone name: pSB1C3
  • Resistance: Cp
  • Copynumber: high

Protein coding:

  • Protein: CaXMT1 [Nucleotide 663 to 1778]
  • Tag: c-terminally fused Strep

Enzymatic activity:

  • EC-number: 2.1.1.158

Cytotoxicity:
This Biobrick is one part of the pathway, which leads to caffeine (the BioBricks BBa_K801074 and BBa_K801075 are further needed). We investigated the toxicity of caffeine and found out, that the growth of yeast strains was inhibited at concentrations of 10 mM. Caffeine with a concentration of 100 mM even showed a lethal effect on yeast cells. Lower concentrations of caffeine (range of micro molar) did not show any growth inhibition or effect.

Safety notes:

Intellectual property:

Corresponding part author/authors:

Source

Source:

  • *Preexisting BioBrick BBa_K801070


Organism:

  • Genesequence derived from coffea arabica
  • Codonoptimized for Saccharomyces cerevisiae
  • Designed for the following Chassis: Saccharomyces cerevisiae

References

Literature references:

  • Uefuji, H., Ogita, S., Yamaguchi, Y., Koizumi, N., and Sano, H. (2003). Molecular cloning and functional characterization of three distinct n-methyltransferases involved in the caffeine biosynthetic pathway in coffee plants. Plant Physiol, 132(1):372–80.

Database references:

  • NCBI