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URL: http://www.addgene.org/65220
Proper Citation: RRID:Addgene_65220
Insert Name: ndmA
Organism: Pseudomonas putida strain CBB5
Bacterial Resistance: Chloramphenicol and Kanamycin
Defining Citation: PMID:23654268
Vector Backbone Description: Backbone Marker:igem parts registry; Backbone Size:2070; Vector Backbone:pSB1C3; Vector Types:Bacterial Expression, Synthetic Biology; Bacterial Resistance:Chloramphenicol and Kanamycin
Comments: The sequence of this plasmid is updated from the GenBank entry for pDCAF3 (KC619530.1) to include an IS5 insertion that occurred in the promoter region of the caffeine demethylation genes during passaging of the strain and a base change in the sequence of the gst9 gene. This plasmid still functions as described in the publication for pDCAF3. Vector constitutively expresses N-demethylase genes under the control of the BBa_J23100 promoter (http://parts.igem.org/Part:BBa_J23100). N-demethylases are only functional when grown at 30°C and require iron supplementation in minimal media. See: Summers, R. M., Louie, T. M., Yu, C.-L., Gakhar, L., Louie, K.C., and Subramanian, M. (2012) Novel, highly specific Ndemethylases enable bacteria to live on caffeine and related purine alkaloids. J. Bacteriol. 194, 2041−2049. For the quantification of caffeine (or other methylxanthine) content it is recommended to grow cells in M9 minimal medium supplemented with 2 g/L glucose and 2 g/L casein (M9GC). Please note that this plasmid may require a unique bacterial strain, so make sure to confirm that you can also obtain the appropriate growth strain. Please contact us at [email protected] or contact our distributors if you have any questions.
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Source: Addgene
Data set from Kravitz, Owen and Kretizer's 2012 paper "Optogenetic identification of striatal projection neuron subtypes during in vivo recordings."
