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

 
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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K801073 short</partinfo>
 
<partinfo>BBa_K801073 short</partinfo>
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<partinfo>BBa_K801073 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K801073 SequenceAndFeatures</partinfo>
  
 +
 +
Composite part of BBa_K801010, BBa_K801070, BBa_K801012
 +
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<br>'''Keywords:''' CaXMT1, TEF2, ADH1, caffeine,
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<!--These keywords are necessary to find your part using a fulltext sarch.-->
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<!--keyword_1, keyword_2, keyword_3, keyword_4, keyword_5-->
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<br>'''Abbreviations:'''
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*CaXMT1 = xanthosine methyltransferase
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<!--*used_abbreviation_2 = full_name_of_used_abbreviations_2-->
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* CaXMT1 = coffea arabica xanthosine N-methyltransferase 1
  
 
===Design Notes===
 
===Design Notes===
−
-
 
  
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'''Related BioBrick:'''
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*Other versions:
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**[https://parts.igem.org/Part:BBa_K801070 BBa_K801070: xanthosine methyltransferase CaXMT1-strep]
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*Related BioBricks:
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** [https://parts.igem.org/Part:BBa_K801071 BBa_K801071: 7-methylxanthine N-methyltransferase CaMXMT1-strep]
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** [https://parts.igem.org/Part:BBa_K801072 BBa_K801072: 3,7-dimethylxanthine N-methyltransferase CaDXMT1-strep]
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** [https://parts.igem.org/Part:BBa_K801073 BBa_K801073: CaXMT1 expression cassette for yeast]
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** [https://parts.igem.org/Part:BBa_K801074 BBa_K801074: CaMXMT1 expression cassette for yeast]
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** [https://parts.igem.org/Part:BBa_K801075 BBa_K801075: CaDXMT1 expression cassette for yeast]
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** [https://parts.igem.org/Part:BBa_K801076 BBa_K801076: Yeast expression cassette for CaXMT1 and CaMXMT1]
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** [https://parts.igem.org/Part:BBa_K801077 BBa_K801077: Caffeine Synthesis Pathway pTEF2-XMT1-tADH1-pTEF1-MXMT1-tADH1-pTEF2-DXMT1-tADH1]
  
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'''Cloning details:'''<br>
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* Designed in RFC10
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<!--*Mutation C889G to delete XbaI restriction site-->
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<!--*Truncation upstream/downstream compared to template, ?explanation?-->
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'''Quality control measures:'''<br>
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* Test digestion using XbaI & PstI
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<!--*Sequencing using primer ?primer_name?/Not yet sequenced-->
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*Part was totally sequenced
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'''Backbone:'''<br>
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* Backbone name: pSB1C3
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* Resistance: Cp
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* Copynumber: high
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'''Protein coding:'''<br>
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* Protein: CaXMT1 [Nucleotide 663 to 1778]
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<!--*The protein has the amino acid replacements ???99??? to ???99???.-->
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<!--*The protein encoded is posttranslationally modified by ???.-->
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* Tag: c-terminally fused Strep
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'''Enzymatic activity:'''
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* EC-number: 2.1.1.158
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 +
'''Cytotoxicity:'''<br>
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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.
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<!--none/not known/cytotoxic for ''organism name''-->
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'''Safety notes:'''<br>
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<!--Known and anticipated sefety issues: none/health_risk/environmental_risk/other_risk-->
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<!--Known and anticipated security issues: none/other.-->
 +
 +
'''Intellectual property:'''
 +
<!--Information on patent situation.-->
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<!--Intellectual property claims made by the authors.-->
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'''Corresponding part author/authors:'''
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<!--https://igem.org/User_Information.cgi?user_id=????/email-->
  
 
===Source===
 
===Source===
  
−
Composite part of BBa_K801010, BBa_K801070, BBa_K801012
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'''Source:'''<br>
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* *Preexisting BioBrick BBa_K801070
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<!--*Commercial system: plasmid name, system name, company name-->
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<!--*Plasmid: p???, provided by ?name_of_person?, ?institute/university?, ?country?-->
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<!--*Preexisting BioBrick ?Bba_number?-->
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<!--*cDNA Clone: ?clone_name?, ?company_name?-->
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<!--*Synthesized by ?company_name?.-->
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<!--'''Forward Primer:'''<br><code>5'- ??? - 3'</code><br>-->
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<!--'''Reverse Primer:'''<br><code>5'- ??? - 3'</code><br>-->
 +
 
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'''Organism:'''<br>
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* Genesequence derived from ''coffea arabica''
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* Codonoptimized for ''Saccharomyces cerevisiae''
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* Designed for the following Chassis: ''Saccharomyces cerevisiae''
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<!--*Statement about functionality in other chassis.-->
  
 
===References===
 
===References===
 +
<!-- Here you find templates to insert references to literature and different databases-->
 +
 +
'''Literature references:'''<br>
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* 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.
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'''Database references:'''<br>
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* NCBI
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<!--*[http://www.ncbi.nlm.nih.gov/nuccore/?accessNr? '''GenBank''': ?title?]-->
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<!--*[http://www.ebi.ac.uk/interpro/IEntry?ac=?accessNr? '''Interpro''': ?title?]-->
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<!--*[http://www.uniprot.org/uniprot/?accessNr? '''Uniprot''': ?title?]-->
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<!--*[http://pfam.sanger.ac.uk/family/?accessNr? '''Pfam:''' ?title?]-->
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<!--*[http://www.rcsb.org/pdb/explore/explore.do?structureId=?accessNR? '''PDB:''' ?tile?]-->
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<!--*[http://www.brenda-enzymes.info/php/result_flat.php4?ecno=?accessNr? '''Branda:''' ?title?]-->

Latest revision as of 10:36, 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:

  • *Preexisting BioBrick BBa_K801070


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