Difference between revisions of "Part:BBa K801073:Design"
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* Genesequence derived from ''coffea arabica'' | * Genesequence derived from ''coffea arabica'' | ||
* Codonoptimized for ''Saccharomyces cerevisiae'' | * Codonoptimized for ''Saccharomyces cerevisiae'' | ||
| − | * Designed for the following Chassis: ''Saccharomyces cerevisiae'' | + | * Designed for the following Chassis: ''Saccharomyces cerevisiae'' |
<!--*Statement about functionality in other chassis.--> | <!--*Statement about functionality in other chassis.--> | ||
Revision as of 10:14, 23 October 2012
CaXMT1 expression cassette for yeast
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 138
- 1000COMPATIBLE WITH RFC[1000]
Composite part of BBa_K801010, BBa_K801070, BBa_K801012
Keywords: CaMXMT1, TEF2 promoter, ADH1 terminator, caffeine, caffeine pathway, caffeine synthesis pathway
Abbreviations:
- CaXMT1 = xanthosine methyltransferase
Design Notes
Related BioBrick:
- Other versions:
- Related BioBricks:
- BBa_K801071: 7-methylxanthine N-methyltransferase CaMXMT1-strep
- BBa_K801072: 3,7-dimethylxanthine N-methyltransferase CaDXMT1-strep
- BBa_K801073: CaXMT1 expression cassette for yeast
- BBa_K801074: CaMXMT1 expression cassette for yeast
- BBa_K801075: CaDXMT1 expression cassette for yeast
- BBa_K801076: Yeast expression cassette for CaXMT1 and CaMXMT1
- BBa_K801077: Caffeine Synthesis Pathway pTEF2-XMT1-tADH1-pTEF1-MXMT1-tADH1-pTEF2-DXMT1-tADH1
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: