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URL: http://www.addgene.org/174374
Proper Citation: RRID:Addgene_174374
Bacterial Resistance: Apramycin
Defining Citation: PMID:34890250
Vector Backbone Description: Backbone Size:7373; Vector Backbone:pUC19; Vector Types:Vector for 2-step knockout by homologous recombination; Bacterial Resistance:Apramycin
Comments: Also known as pSelAct. For cloning in homology arms, can use Gateway, restriction-ligation, or homology-based cloning such as Gibson/Infusion cloning. Please cite at minimum paper 1) and a secondary paper that is most appropriate to your system of interest (2 or 3). 1. First development of pSelAct for knockouts in Rhodococcus equi: Geize, R. van der, Jong, W. de, Hessels, G. I., Grommen, A. W. F., Jacobs, A. A. C., and Dijkhuizen, L. 2008. A novel method to generate unmarked gene deletions in the intracellular pathogen Rhodococcus equi using 5-fluorocytosine conditional lethality. Nucleic Acids Res. 36:e151 2. First development of pSelAct with Gateway cassette for knockouts in Rhodococcus fasicans: Savory, E. A., Weisberg, A. J., Stevens, D. M., Creason, A. L., Fuller, S. L., Pearce, E. M., and Chang, J. H. 2020. Phytopathogenic Rhodococcus Have Diverse Plasmids With Few Conserved Virulence Functions. Front Microbiol. 11:1022 3. Later referred to as pSelAct-KO after optimization for knockouts in Clavibacter: Stevens, D. M., Tang, A., and Coaker, G. 2021. A Genetic Toolkit for Investigating Clavibacter Species: Markerless Deletion, Permissive Site Identification, and an Integrative Plasmid. Mol Plant-microbe Interactions. 34:1336–1345
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Source: Addgene