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

Revision as of 12:59, 3 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: CaMXMT1, TEF2 promoter, ADH1 terminator, caffeine, caffeine pathway, caffeine synthesis pathway

Abbreviations:

CaXMT1 = xanthosine methyltransferase

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:

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 '''Literature references:'''<br>
-<!--*[http://www.ncbi.nlm.nih.gov/pubmed/?PMID? '''Pubmed:''' ?Author(s)?, ?year?: ?title?]-->
+* 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>