Difference between revisions of "Part:BBa K801075:Design"

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<br>'''Keywords:'''
 
<br>'''Keywords:'''
 
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caffeine, TEF1, expression cassette
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caffeine, TEF2, expression cassette
  
 
<br>'''Abbreviations:'''
 
<br>'''Abbreviations:'''
 
<!--*used_abbreviation_1 = full_name_of_used_abbreviations_1-->
 
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* CaDXMT1 = coffea arabica 3,7-dimethylxanthine N-methyltransferase 1
  
 
===Design Notes===
 
===Design Notes===
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'''Organism:'''<br>
 
'''Organism:'''<br>
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* Genesequence derived from ''coffea arabica''
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* Genesequence derived from ''Coffea arabica''
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* codonoptimized for ''saccharomyces cerevisiae''
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* codonoptimized for ''Saccharomyces cerevisiae''
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* designed for ''saccharomyces cerevisiae''
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* designed for ''Saccharomyces cerevisiae''
 
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Latest revision as of 10:31, 24 October 2012

CaDXMT1 expression cassette for yeast


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1650
    Illegal BglII site found at 1746
  • 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_K801072 and BBa_K801012
Keywords: caffeine, TEF2, expression cassette


Abbreviations:

  • CaDXMT1 = coffea arabica 3,7-dimethylxanthine N-methyltransferase 1

Design Notes

Related BioBrick: BBa_K801072 BBa_K801077

Quality control measures:

  • Test digest has been performed with Xba1 and Pst1 (see mainpage)
  • Part was totally sequenced

Backbone:

  • Backbone name: pSB1C3'
  • Resistance Cp

Protein coding:

  • CaDXMT1

Enzymatic activity: EC-number [http://enzyme.expasy.org/EC/2.1.1.160 2.1.1.160 ]

Cytotoxicity:
This Biobrick is one part of the pathway, which leads to caffeine (the BioBricks BBa_K801073 and BBa_K801074 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_K801072


Organism:

  • Genesequence derived from Coffea arabica
  • codonoptimized for Saccharomyces cerevisiae
  • designed for Saccharomyces cerevisiae

References

Literature references:

  • [http://www.ncbi.nlm.nih.gov/pubmed/12746542 Pubmed: 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.]
  • [http://www.ncbi.nlm.nih.gov/pubmed/16247553 Pubmed: Uefuji H., Ogita S., Yamaguchi Y., Koizumi N. and Sano H., 2005: Caffeine production in tobacco plants by simultaneous expression of three coffee N-methyltrasferases and its potential as a pest repellant.]
  • [http://www.ncbi.nlm.nih.gov/pubmed/16925551 Pubmed: Kuranda K, Leberre V, Sokol S, Palamarczyk G, François J., 2006: Investigating the caffeine effects in the yeast Saccharomyces cerevisiae brings new insights into the connection between TOR, PKC and Ras/cAMP signalling pathways.]

Database references:

  • NCBI