Difference between revisions of "Part:BBa K801075:Design"
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<br>'''Keywords:''' | <br>'''Keywords:''' | ||
<!--These keywords are necessary to find your part using a fulltext sarch.--> | <!--These keywords are necessary to find your part using a fulltext sarch.--> | ||
| − | caffeine, | + | caffeine, TEF2, expression cassette |
<br>'''Abbreviations:''' | <br>'''Abbreviations:''' | ||
<!--*used_abbreviation_1 = full_name_of_used_abbreviations_1--> | <!--*used_abbreviation_1 = full_name_of_used_abbreviations_1--> | ||
<!--*used_abbreviation_2 = full_name_of_used_abbreviations_2--> | <!--*used_abbreviation_2 = full_name_of_used_abbreviations_2--> | ||
| + | * CaDXMT1 = coffea arabica 3,7-dimethylxanthine N-methyltransferase 1 | ||
===Design Notes=== | ===Design Notes=== | ||
'''Related BioBrick:''' | '''Related BioBrick:''' | ||
| + | [https://parts.igem.org/Part:BBa_K801075 BBa_K801072] | ||
| + | [https://parts.igem.org/Part:BBa_K801077 BBa_K801077] | ||
<!--*Other versions:[https://parts.igem.org/wiki/index.php?title=Part:BBa_?????? BBa_??????: Name_of_part] --> | <!--*Other versions:[https://parts.igem.org/wiki/index.php?title=Part:BBa_?????? BBa_??????: Name_of_part] --> | ||
<!--*Related BioBricks:[https://parts.igem.org/wiki/index.php?title=Part:BBa_?????? BBa_??????: Name_of_part] | <!--*Related BioBricks:[https://parts.igem.org/wiki/index.php?title=Part:BBa_?????? BBa_??????: Name_of_part] | ||
'''Cloning details:'''<br> | '''Cloning details:'''<br> | ||
| + | * Designed in RFC10 | ||
<!--*Designed in RFC10/RFC23/RFC25/RFC25_N-part--> | <!--*Designed in RFC10/RFC23/RFC25/RFC25_N-part--> | ||
<!--*Mutation C889G to delete XbaI restriction site--> | <!--*Mutation C889G to delete XbaI restriction site--> | ||
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'''Quality control measures:'''<br> | '''Quality control measures:'''<br> | ||
| + | * Test digest has been performed with Xba1 and Pst1 (see mainpage) | ||
| + | * Part was totally sequenced | ||
<!--*Test digestion using ?enzyme1? & ?enzyme2?/Not yet performed--> | <!--*Test digestion using ?enzyme1? & ?enzyme2?/Not yet performed--> | ||
<!--*Sequencing using primer ?primer_name?/Not yet sequenced--> | <!--*Sequencing using primer ?primer_name?/Not yet sequenced--> | ||
| Line 32: | Line 38: | ||
'''Backbone:'''<br> | '''Backbone:'''<br> | ||
| − | *Backbone name: pSB1C3' | + | * Backbone name: pSB1C3' |
| + | * Resistance Cp | ||
<!--*Resistance: Amp/Cp/Kan/Tet--> | <!--*Resistance: Amp/Cp/Kan/Tet--> | ||
<!--*Copynumber: low/medium/high--> | <!--*Copynumber: low/medium/high--> | ||
'''Protein coding:'''<br> | '''Protein coding:'''<br> | ||
| + | * CaDXMT1 | ||
<!--*Protein: ?Name_of_gene_product? [Nucleotide 1 to ???]--> | <!--*Protein: ?Name_of_gene_product? [Nucleotide 1 to ???]--> | ||
<!--*The protein has the amino acid replacements ???99??? to ???99???.--> | <!--*The protein has the amino acid replacements ???99??? to ???99???.--> | ||
| Line 43: | Line 51: | ||
'''Enzymatic activity:''' | '''Enzymatic activity:''' | ||
| + | EC-number [http://enzyme.expasy.org/EC/2.1.1.160 2.1.1.160 ] | ||
<!--none/EC-number ?.?.?.?--> | <!--none/EC-number ?.?.?.?--> | ||
| Line 75: | Line 84: | ||
'''Organism:'''<br> | '''Organism:'''<br> | ||
| − | * Genesequence derived from '' | + | * Genesequence derived from ''Coffea arabica'' |
| − | * codonoptimized for '' | + | * codonoptimized for ''Saccharomyces cerevisiae'' |
| − | * designed for '' | + | * designed for ''Saccharomyces cerevisiae'' |
<!--*Genesequence derived from ''?organism_name?''--> | <!--*Genesequence derived from ''?organism_name?''--> | ||
<!--*Codonoptimized for ''?organism_name?''--> | <!--*Codonoptimized for ''?organism_name?''--> | ||
Latest revision as of 10:31, 24 October 2012
CaDXMT1 expression cassette for yeast
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1650
Illegal BglII site found at 1746 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 138
- 1000COMPATIBLE 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