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Species: Stackebrandtia nassauensis
Genetic Insert: SnaA
Vector Backbone Description: Vector Backbone:pOSV801; Vector Types:Bacterial Expression; Bacterial Resistance:Apramycin
Defining Citation: PMID:38947204
Comments: encodes biosynthetic genes for threopeptin, including SnaA, SnaB, SnaC, SnaD, SnaE, SnaO, SnaT1, SnaT2
Proper citation: RRID:Addgene_225055 Copy
Genetic Insert: apramycin resistance gene
Vector Backbone Description: Vector Backbone:pSGAb; Vector Types:Bacterial Expression; Bacterial Resistance:Apramycin
Proper citation: RRID:Addgene_226171 Copy
Species: Synthetic
Genetic Insert: T7RNAP Circuit
Vector Backbone Description: Vector Backbone:Custom; Vector Types:Bacterial Expression; Bacterial Resistance:Apramycin
Defining Citation: PMID:35366417
Proper citation: RRID:Addgene_199993 Copy
Species: Synthetic
Genetic Insert: sSpCas9
Vector Backbone Description: Vector Backbone:pKC1139; Vector Types:Bacterial Expression, CRISPR; Bacterial Resistance:Apramycin
Defining Citation: PMID:25458909
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
Defining Citation: PMID:25458909
Proper citation: RRID:Addgene_61737 Copy
Vector Backbone Description: Vector Backbone:pIJ8668; Vector Types:Bacterial Expression, Streptomyces expression; Bacterial Resistance:Apramycin
Proper citation: RRID:Addgene_78576 Copy
Species: E. coli
Genetic Insert: aac
Vector Backbone Description: Backbone Size:2140; Vector Backbone:pSG5; Vector Types:; Bacterial Resistance:Apramycin
Defining Citation: PMID:30191290
Proper citation: RRID:Addgene_69615 Copy
Species: E. coli
Genetic Insert: aac
Vector Backbone Description: Backbone Size:2140; Vector Backbone:pSG5; Vector Types:Bacterial Expression; Bacterial Resistance:Apramycin
Defining Citation: PMID:30191290
Proper citation: RRID:Addgene_69994 Copy
Vector Backbone Description: Backbone Size:6440; Vector Backbone:pUC19; Vector Types:Expression vector for targeted integration; Bacterial Resistance:Apramycin
Defining Citation: PMID:34890250
Comments: For cloning in homology arms, can use Gateway, restriction ligation, or homology-based cloning such as Gibson/Infusion cloning. eGFP can be swapped for your gene of interest or can be fused (recommend homology base cloning).
Original pSelAct backbone was developed here:
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
pSelAct-Express was develop and tested here:
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_174375 Copy
Species: Synthetic
Genetic Insert: attL
Vector Backbone Description: Backbone Marker:Bierman M. et al. Gene 1992 Plasmid cloning vectors for the conjugal transfer of DNA from Escherichia coli to Streptomyces spp.; Backbone Size:3469; Vector Backbone:pOJ260; Vector Types:Bacterial Expression; Bacterial Resistance:Apramycin
Defining Citation: PMID:35456877
Proper citation: RRID:Addgene_172193 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
Defining Citation: PMID:25739462
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
Vector Backbone Description: Vector Backbone:pSynORI; Vector Types:Bacterial Expression; Bacterial Resistance:Apramycin
Defining Citation: PMID:41629321
Comments: J23119-Repressor (F3m1)-RNA Primer (-68nt)-Target (F3m1)-J23110-parDE-parCBA-AprR
Proper citation: RRID:Addgene_253209 Copy
Genetic Insert: SceI
Vector Backbone Description: Vector Backbone:pSW-Apr; Vector Types:Bacterial Expression; Bacterial Resistance:Apramycin
Defining Citation: PMID:36216579
Proper citation: RRID:Addgene_252219 Copy
Species: Acinetobacter baumannii
Genetic Insert: mltB
Vector Backbone Description: Backbone Marker:Addgene; Vector Backbone:pSuiAb; Vector Types:Bacterial Expression; Bacterial Resistance:Apramycin
Proper citation: RRID:Addgene_253612 Copy
Genetic Insert: gRNA targetting desD in Streptomyces
Vector Backbone Description: Backbone Marker:Addgene plasmid #125689 (Weber lab); Backbone Size:12247; Vector Backbone:pCRISPR-cBEST; Vector Types:CRISPR; Bacterial Resistance:Apramycin
Defining Citation: PMID:41942569
Proper citation: RRID:Addgene_219650 Copy
Genetic Insert: Protospacer targeting desD in Streptomyces
Vector Backbone Description: Backbone Marker:Addgene plasmid #125689 (Weber lab); Vector Backbone:pCRISPR-cBEST; Vector Types:CRISPR; Bacterial Resistance:Apramycin
Defining Citation: PMID:41942569
Proper citation: RRID:Addgene_219651 Copy
Species: Synthetic
Genetic Insert: CRISPRt transposon with gRNA expression cassette (mScar-T03 guide)
Vector Backbone Description: Vector Backbone:R6K_; Vector Types:Bacterial Expression, Synthetic Biology, CRISPR Associated Transposition (CAST); Bacterial Resistance:Apramycin
Comments: Please visit https://doi.org/10.1101/2024.03.01.582922 for bioRxiv preprint.
Proper citation: RRID:Addgene_225838 Copy
Species: Synthetic
Genetic Insert: CRISPRtOE transposon with gRNA expression cassette (no guide) and synthetic promoter H
Vector Backbone Description: Vector Backbone:R6K_; Vector Types:Bacterial Expression, Synthetic Biology, CRISPR Associated Transposition (CAST); Bacterial Resistance:Apramycin
Comments: Please visit https://doi.org/10.1101/2024.03.01.582922 for bioRxiv preprint.
Proper citation: RRID:Addgene_225854 Copy
Species: Synthetic
Genetic Insert: CRISPRtOE transposon with gRNA expression cassette (no guide) and no outward facing promoter
Vector Backbone Description: Vector Backbone:R6K_; Vector Types:Bacterial Expression, Synthetic Biology, CRISPR Associated Transposition (CAST); Bacterial Resistance:Apramycin
Comments: Please visit https://doi.org/10.1101/2024.03.01.582922 for bioRxiv preprint.
Proper citation: RRID:Addgene_225853 Copy
Species: Synthetic
Genetic Insert: Tn7 transposon with mScarlet-I expression
Vector Backbone Description: Vector Backbone:R6K_; Vector Types:Bacterial Expression, Synthetic Biology, Tn7 Transposition; Bacterial Resistance:Apramycin
Comments: Please visit https://doi.org/10.1101/2024.03.01.582922 for bioRxiv preprint.
Proper citation: RRID:Addgene_225866 Copy
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