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

(→‎Design Notes)
(→‎Design Notes)
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'''Cloning details:'''<br>
 
'''Cloning details:'''<br>
−
<!--*Designed in RFC10/RFC23/RFC25/RFC25_N-part-->
+
* Designed in RFC10
 
<!--*Mutation C889G to delete XbaI restriction site-->
 
<!--*Mutation C889G to delete XbaI restriction site-->
 
<!--*Truncation upstream/downstream compared to template, ?explanation?-->
 
<!--*Truncation upstream/downstream compared to template, ?explanation?-->
  
 
'''Quality control measures:'''<br>
 
'''Quality control measures:'''<br>
−
<!--*Test digestion using ?enzyme1? & ?enzyme2?/Not yet performed-->
+
* Test digestion using XbaI & PstI
 
<!--*Sequencing using primer ?primer_name?/Not yet sequenced-->
 
<!--*Sequencing using primer ?primer_name?/Not yet sequenced-->
 
*Part was totally sequenced
 
*Part was totally sequenced
  
 
'''Backbone:'''<br>
 
'''Backbone:'''<br>
−
*Backbone name: pSB1C3
+
* Backbone name: pSB1C3
−
<!--*Resistance: Amp/Cp/Kan/Tet-->
+
* Resistance: Cp
−
<!--*Copynumber: low/medium/high-->
+
* Copynumber: high
  
 
'''Protein coding:'''<br>
 
'''Protein coding:'''<br>
−
<!--*Protein: ?Name_of_gene_product? [Nucleotide 1 to ???]-->
+
* Protein: CaXMT1 [Nucleotide 663 to 1778]
 
<!--*The protein has the amino acid replacements ???99??? to ???99???.-->
 
<!--*The protein has the amino acid replacements ???99??? to ???99???.-->
 
<!--*The protein encoded is posttranslationally modified by ???.-->
 
<!--*The protein encoded is posttranslationally modified by ???.-->
−
<!--*Tag: n-terminally fused/c-terminally fused His5/His6/Strep/Flag/other-->
+
* Tag: c-terminally fused Strep
  
 
'''Enzymatic activity:'''
 
'''Enzymatic activity:'''
−
<!--none/EC-number ?.?.?.?-->
+
* EC-number: 2.1.1.158
  
 
'''Cytotoxicity:'''<br>
 
'''Cytotoxicity:'''<br>

Revision as of 12:56, 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:


Organism:


References

Literature references:

Database references: