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
  
<br>'''Keywords:''' CaMXMT1, TEF2 promoter, ADH1 terminator, caffeine, caffeine pathway, caffeine synthesis pathway
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<br>'''Keywords:''' CaXMT1, TEF2, ADH1, caffeine,
 
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*CaXMT1 = xanthosine methyltransferase
 
*CaXMT1 = xanthosine methyltransferase
 
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* CaXMT1 = coffea arabica xanthosine N-methyltransferase 1
  
 
===Design Notes===
 
===Design Notes===

Revision as of 10:34, 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:

  • CaXMT1 was synthesized by IDT


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