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Species:
Genetic Insert:
Vector Backbone Description: Vector Backbone:pACEC3GH; Vector Types:Bacterial Expression; Bacterial Resistance:Apramycin
References:
Comments:
Proper citation: RRID:Addgene_110085 Copy
Species:
Genetic Insert:
Vector Backbone Description: Vector Backbone:pKO; Vector Types:Bacterial Knockout Generation; Bacterial Resistance:Apramycin
References:
Comments:
Proper citation: RRID:Addgene_110088 Copy
Species: Synthetic
Genetic Insert: mCherry, aac(3)IV, tsr
Vector Backbone Description: Vector Backbone:pSG5; Vector Types:Bacterial Expression; Bacterial Resistance:Apramycin
References:
Comments: Mobilizable, stable replicating expression vector for Streptomyces, strong constitutive SF14* promoter, codon optimized Strep-tagII-mCherry gene, tsr, aac(3)IV
Proper citation: RRID:Addgene_115683 Copy
Species: Streptomyces
Genetic Insert: gusA, aac(3)IV, tsr
Vector Backbone Description: Vector Backbone:pSG5; Vector Types:Gene knock out; Bacterial Resistance:Apramycin
References:
Comments: Thermosensitive replication (< 37°C) in Streptomyces, Detection of single/double cross over by X-gluc colour reaction , aac(3)IV
Proper citation: RRID:Addgene_115678 Copy
Species: Synthetic
Genetic Insert: sSpCas9
Vector Backbone Description: Vector Backbone:pKC1139; Vector Types:Bacterial Expression, CRISPR; Bacterial Resistance:Apramycin
References:
Comments:
Proper citation: RRID:Addgene_61736 Copy
Species: Synthetic
Genetic Insert: sSpCas9
Vector Backbone Description: Vector Backbone:pKC1139; Vector Types:Bacterial Expression, CRISPR; Bacterial Resistance:Apramycin
References:
Comments:
Proper citation: RRID:Addgene_61737 Copy
Species: artificially synthesized, codon optimized gene from Strep pyogenes
Genetic Insert: Scocas9
Vector Backbone Description: Vector Backbone:pKC1139; Vector Types:Bacterial Expression, CRISPR, Bacterial Genome Editing; Bacterial Resistance:Apramycin
References:
Comments: More information is provided in the supplemental files. Please note that the sequence of the backbone (pKC1139) was obtained via Sanger sequencing and several regions may have discrepancies due to high GC content. These discrepancies have no functional consequences.
Proper citation: RRID:Addgene_62552 Copy
Species: Streptomyces
Genetic Insert: Streptomyces codon optimized the FnCas12a mutant EP16
Vector Backbone Description: Vector Backbone:pCRISPomyces-2; Vector Types:CRISPR; Bacterial Resistance:Apramycin
References:
Comments:
Proper citation: RRID:Addgene_177875 Copy
Species:
Genetic Insert:
Vector Backbone Description: Backbone Size:7373; Vector Backbone:pUC19; Vector Types:Vector for 2-step knockout by homologous recombination; Bacterial Resistance:Apramycin
References:
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
Proper citation: RRID:Addgene_174374 Copy
Species: Homo sapiens
Genetic Insert: Ubiquitin
Vector Backbone Description: Backbone Size:3896; Vector Backbone:pBAD; Vector Types:Bacterial Expression; Bacterial Resistance:Apramycin
References:
Comments:
Proper citation: RRID:Addgene_174517 Copy
Species: M. smegmatis
Genetic Insert: MSMEG_0948
Vector Backbone Description: Vector Backbone:pMSR:Dendra; Vector Types:Bacterial Expression, Integrating Mycobacterial Expression Vector attL5. T7 expression in E. coli; Bacterial Resistance:Apramycin
References:
Comments: 5' cloning site: AseI (destroyed), 3' cloning site: HindIII (not destroyed)
Proper citation: RRID:Addgene_174952 Copy
Species: M. smegmatis
Genetic Insert: MSMEG_0952
Vector Backbone Description: Vector Backbone:pMSR:Dendra; Vector Types:Bacterial Expression, Integrating Mycobacterial Expression Vector attL5. T7 expression in E. coli; Bacterial Resistance:Apramycin
References:
Comments: 5' cloning site: AseI (destroyed), 3' cloning site: HindIII (not destroyed)
Proper citation: RRID:Addgene_174954 Copy
Species: M. smegmatis
Genetic Insert: MSMEG_0953
Vector Backbone Description: Vector Backbone:pMSR:Dendra; Vector Types:Bacterial Expression, Integrating Mycobacterial Expression Vector attL5. T7 expression in E. coli; Bacterial Resistance:Apramycin
References:
Comments: 5' cloning site: AseI (destroyed), 3' cloning site: HindIII (not destroyed)
Proper citation: RRID:Addgene_174955 Copy
Species: M. smegmatis
Genetic Insert: MSMEG_0954
Vector Backbone Description: Vector Backbone:pMSR:Dendra; Vector Types:Bacterial Expression, Integrating Mycobacterial Expression Vector attL5. T7 expression in E. coli; Bacterial Resistance:Apramycin
References:
Comments: 5' cloning site: AseI (destroyed), 3' cloning site: HindIII (not destroyed)
Proper citation: RRID:Addgene_174956 Copy
Species: M. smegmatis
Genetic Insert: MSMEG_0956
Vector Backbone Description: Vector Backbone:pMSR:Dendra; Vector Types:Bacterial Expression, Integrating Mycobacterial Expression Vector attL5. T7 expression in E. coli; Bacterial Resistance:Apramycin
References:
Comments: 5' cloning site: AseI (destroyed), 3' cloning site: HindIII (not destroyed)
Proper citation: RRID:Addgene_174957 Copy
Species: M. smegmatis
Genetic Insert: MSMEG_0970
Vector Backbone Description: Vector Backbone:pMSR:Dendra; Vector Types:Bacterial Expression, Integrating Mycobacterial Expression Vector attL5. T7 expression in E. coli; Bacterial Resistance:Apramycin
References:
Comments: 5' cloning site: AseI (destroyed), 3' cloning site: HindIII (not destroyed)
Proper citation: RRID:Addgene_174959 Copy
Species: M. smegmatis
Genetic Insert: MSMEG_0946
Vector Backbone Description: Vector Backbone:pMSR:Dendra; Vector Types:Bacterial Expression, Integrating Mycobacterial Expression Vector attL5. T7 expression in E. coli; Bacterial Resistance:Apramycin
References:
Comments: 5' cloning site: AseI (destroyed), 3' cloning site: HindIII (not destroyed)
Proper citation: RRID:Addgene_174950 Copy
Species: M. smegmatis
Genetic Insert: MSMEG_0943
Vector Backbone Description: Vector Backbone:pMSR:Dendra; Vector Types:Bacterial Expression, Integrating Mycobacterial Expression Vector attL5. T7 expression in E. coli; Bacterial Resistance:Apramycin
References:
Comments: 5' cloning site: AseI (destroyed), 3' cloning site: HindIII (not destroyed)
Proper citation: RRID:Addgene_174949 Copy
Species: M. smegmatis
Genetic Insert: MSMEG_0933
Vector Backbone Description: Vector Backbone:pMSR:Dendra; Vector Types:Bacterial Expression, Integrating Mycobacterial Expression Vector attL5. T7 expression in E. coli; Bacterial Resistance:Apramycin
References:
Comments: 5' cloning site: AseI (destroyed), 3' cloning site: HindIII (not destroyed)
Proper citation: RRID:Addgene_174941 Copy
Species: M. smegmatis
Genetic Insert: MSMEG_0934
Vector Backbone Description: Vector Backbone:pMSR:Dendra; Vector Types:Bacterial Expression, Integrating Mycobacterial Expression Vector attL5. T7 expression in E. coli; Bacterial Resistance:Apramycin
References:
Comments: 5' cloning site: AseI (destroyed), 3' cloning site: HindIII (not destroyed)
Proper citation: RRID:Addgene_174942 Copy
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