title: The Standard European Vector Architecture (SEVA) plasmid toolkit.
abstract: The Standard European Vector Architecture (SEVA) toolkit is a simple and powerful resource for constructing optimal plasmid vectors based on a backbone and three interchangeable modules flanked by uncommon restriction sites. Functional modules encode several origins of replication, diverse antibiotic selection markers, and a variety of cargoes with different applications. The backbone and DNA modules have been minimized and edited for flaws in their sequence and/or functionality. A protocol for the utilization of the SEVA platform to construct transcriptional and translational fusions between a promoter under study (the arsenic-responsive Pars of Pseudomonas putida KT2440) and the reporter lacZ gene is described. The resulting plasmid collection was instrumental to measure and compare the β-galactosidase activity that report gene expression (i.e., transcription and translation) in different genetic backgrounds.
authors.affiliation: Department of Environmental Biology, Centro de Investigaciones Biológicas, Ramiro de Maeztu 9, 28040, Madrid, Spain.
authors.forename: Gonzalo
authors.initials: G
authors.lastname: Durante-Rodríguez
authors.forename: Víctor
authors.initials: V
authors.lastname: de Lorenzo
authors.forename: Esteban
authors.initials: E
authors.lastname: Martínez-García
mesh_terms: D001483:Base Sequence; D003001:Cloning, Molecular; D017931:DNA Primers / Q000378:metabolism; D005060:Europe; D005822:Genetic Vectors / Q000378:metabolism*; D008967:Molecular Biology / Q000379:methods*; D008969:Molecular Sequence Data; D009876:Operon / Q000235:genetics; D010957:Plasmids / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D016958:Pseudomonas putida / Q000235:genetics; D012015:Reference Standards; D001616:beta-Galactosidase / Q000378:metabolism
chemical_list: D017931:DNA Primers; D001616:beta-Galactosidase