Difference between revisions of "Part:BBa K801075:Design"

Revision as of 10:40, 23 October 2012

CaDXMT1 expression cassette for yeast

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 1650
Illegal BglII site found at 1746
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_K801072 and BBa_K801012
Keywords: caffeine, TEF1, expression cassette

Abbreviations:

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

@@ Line 18: / Line 18: @@
 '''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] -->
 <!--*Related BioBricks:[https://parts.igem.org/wiki/index.php?title=Part:BBa_?????? BBa_??????: Name_of_part]
 '''Cloning details:'''<br>
+* Designed in RFC10
 <!--*Designed in RFC10/RFC23/RFC25/RFC25_N-part-->
 <!--*Mutation C889G to delete XbaI restriction site-->
@@ Line 27: / Line 30: @@
 '''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-->
 <!--*Sequencing using primer ?primer_name?/Not yet sequenced-->
@@ Line 32: / Line 37: @@
 '''Backbone:'''<br>
-*Backbone name: pSB1C3'
+* Backbone name: pSB1C3'
+* Resistance Cp
 <!--*Resistance: Amp/Cp/Kan/Tet-->
 <!--*Copynumber: low/medium/high-->
 '''Protein coding:'''<br>
+* CaDXMT1
 <!--*Protein: ?Name_of_gene_product? [Nucleotide 1 to ???]-->
 <!--*The protein has the amino acid replacements ???99??? to ???99???.-->
@@ Line 43: / Line 50: @@
 '''Enzymatic activity:'''
+EC-number [http://enzyme.expasy.org/EC/2.1.1.160  2.1.1.160 ]
 <!--none/EC-number ?.?.?.?-->