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http://www.addgene.org/105134

Species: C. difficile
Genetic Insert: Upstream & Downstream pyrE deletion region
Vector Backbone Description: Backbone Size:6324; Vector Backbone:pJS116; Vector Types:E. coli - C. difficile shuttle vector; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:29116155

Proper citation: RRID:Addgene_105134 Copy   

•   Source: Addgene

http://www.addgene.org/105133

Species: C. difficile
Genetic Insert: Upstream & Downstream pyrE deletion region
Vector Backbone Description: Backbone Size:6297; Vector Backbone:pMTL84151; Vector Types:E. coli - C. difficile shuttle vector; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:29116155

Proper citation: RRID:Addgene_105133 Copy   

•   Source: Addgene

http://www.addgene.org/105497

Species: C. difficile / synthetic
Genetic Insert: SmBiT/HupA-LgBiT
Vector Backbone Description: Backbone Marker:R.P. Fagan, University of Sheffield; Vector Backbone:pRPF185; Vector Types:ATc-dependent expression in C. difficile; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:30633871
Comments: Please visit https://www.biorxiv.org/content/early/2018/09/27/426809 for bioRxiv preprint.

Proper citation: RRID:Addgene_105497 Copy   

•   Source: Addgene

http://www.addgene.org/105496

Species: C. difficile / synthetic
Genetic Insert: HupA-SmBiT/LgBiT
Vector Backbone Description: Backbone Marker:R.P. Fagan, University of Sheffield; Vector Backbone:pRPF185; Vector Types:ATc-dependent expression in C. difficile; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:30633871
Comments: Please visit https://www.biorxiv.org/content/early/2018/09/27/426809 for bioRxiv preprint.

Proper citation: RRID:Addgene_105496 Copy   

•   Source: Addgene

http://www.addgene.org/183151

Vector Backbone Description: Vector Backbone:pYL322d1; Vector Types:Plant Expression; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:35179286

Proper citation: RRID:Addgene_183151 Copy   

•   Source: Addgene

http://www.addgene.org/18876

Species: E. coli
Genetic Insert: polyphosphate kinase
Vector Backbone Description: Backbone Size:0; Vector Backbone:pMMB206; Vector Types:Bacterial Expression, Synthetic Biology; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:9143103

Proper citation: RRID:Addgene_18876 Copy   

•   Source: Addgene

http://www.addgene.org/18878

Vector Backbone Description: Backbone Size:6573; Vector Backbone:pYUB1049; Vector Types:Bacterial Expression; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:17949993
Comments: This is a Gateway destination vector. It requires a ccdB survival strain of E. coli for propagation of the plasmid, but once a gene of interest has been cloned in (via the LR reaction) it will grow in standard E. coli strains (eg. DH5alpha, BL21). This is a shuttle vector, designed for expression of the inserted gene in Mycobacterium smegmatis as well as allowing cloning using E. coli.

Proper citation: RRID:Addgene_18878 Copy   

•   Source: Addgene

http://www.addgene.org/19001

Species: E. coli
Genetic Insert: arabinose transporter araE under control of Ptac
Vector Backbone Description: Backbone Size:0; Vector Backbone:pMMB207; Vector Types:Bacterial Expression, Synthetic Biology; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:11092865

Proper citation: RRID:Addgene_19001 Copy   

•   Source: Addgene

http://www.addgene.org/19002

Species: E. herbicola
Genetic Insert: crtE, crtB
Vector Backbone Description: Backbone Marker:Cunningham et al., 1994; Backbone Size:0; Vector Backbone:pAC-LYC; Vector Types:Bacterial Expression, Synthetic Biology; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:11676567

Proper citation: RRID:Addgene_19002 Copy   

•   Source: Addgene

http://www.addgene.org/110574

Species: Synthetic
Genetic Insert: T7 promoter with RBS
Vector Backbone Description: Vector Backbone:ColE1; Vector Types:Synthetic Biology; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:29608282

Proper citation: RRID:Addgene_110574 Copy   

•   Source: Addgene

http://www.addgene.org/110575

Species: Synthetic
Genetic Insert: CP25 Promoter with RBS
Vector Backbone Description: Vector Backbone:ColE1; Vector Types:Synthetic Biology; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:29608282

Proper citation: RRID:Addgene_110575 Copy   

•   Source: Addgene

http://www.addgene.org/110578

Species: Synthetic
Genetic Insert: CP32 Promoter with RBS
Vector Backbone Description: Vector Backbone:ColE1; Vector Types:Synthetic Biology; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:29608282

Proper citation: RRID:Addgene_110578 Copy   

•   Source: Addgene

http://www.addgene.org/110577

Species: Synthetic
Genetic Insert: CP12b Promoter with RBS
Vector Backbone Description: Vector Backbone:ColE1; Vector Types:Synthetic Biology; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:29608282

Proper citation: RRID:Addgene_110577 Copy   

•   Source: Addgene

http://www.addgene.org/110547

Species: Synthetic
Genetic Insert: dCas9
Vector Backbone Description: Backbone Marker:Novagen/Agilent; Backbone Size:2412; Vector Backbone:pACYC; Vector Types:Bacterial Expression; Bacterial Resistance:Chloramphenicol
Comments: pIM2432

Proper citation: RRID:Addgene_110547 Copy   

•   Source: Addgene

http://www.addgene.org/110592

Species: Synthetic
Genetic Insert: rpoC
Vector Backbone Description: Vector Backbone:ColE1; Vector Types:Synthetic Biology; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:29608282

Proper citation: RRID:Addgene_110592 Copy   

•   Source: Addgene

http://www.addgene.org/110582

Species: Synthetic
Genetic Insert: Lac promoter with lac operator and RBS
Vector Backbone Description: Vector Backbone:ColE1; Vector Types:Synthetic Biology; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:29608282

Proper citation: RRID:Addgene_110582 Copy   

•   Source: Addgene

http://www.addgene.org/110742

Vector Backbone Description: Backbone Size:2979; Vector Backbone:pUCDM; Vector Types:Insect Expression; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:17117155

Proper citation: RRID:Addgene_110742 Copy   

•   Source: Addgene

http://www.addgene.org/111656

Species: Synthetic
Genetic Insert: TetR and xCas9
Vector Backbone Description: Vector Backbone:p15A origin; Vector Types:Bacterial Expression, Synthetic Biology; Bacterial Resistance:Chloramphenicol

Proper citation: RRID:Addgene_111656 Copy   

•   Source: Addgene

http://www.addgene.org/111986

Vector Backbone Description: Backbone Size:2787; Vector Backbone:pACYCDuet-1; Vector Types:Bacterial Expression; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:29724956

Proper citation: RRID:Addgene_111986 Copy   

•   Source: Addgene

http://www.addgene.org/25769

Vector Backbone Description: Backbone Marker:N/A; Backbone Size:8673; Vector Backbone:pMAK700, pMAK705, pBS-TS; Vector Types:Bacterial Expression; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:9335267
Comments: pKOV is identical to the pKO3 vector described in J. Bacteriology 179: 6228-6237, except for the addition of a 3kb stuffer sequence in the multiple cloning site. This stuffer permits (i) directional cloning using NotI and BamHI and (ii) clean separation of doubly cut vector from singly cut contaminants when using this pair of enzymes. The pKOV cloning site is: 5' - SmaI - NotI - SmaI- stuffer - BamHI - SalI - 3'. BamHI and SalI are not used together, nor is SmaI used with NotI. BglII & BclI cut ends are compatible with BamHI. PmeI & SwaI are compatible with SmaI. NOTE: pKOV has a temperature sensitive pSC101 replication origin. To recover the plasmid, strains harboring the plasmid must be grown at 30 deg C under chloramphenicol selection. Gene replacement: Mutant alleles cloned into the pKOV gene replacement vector are electroporated into recombination proficient strains (eg. EMG2) and allowed to recover for 1 h at 30 deg C. The cells are plated on prewarmed chloramphenicol/LB plates and incubated at 42 deg C. To measure the integration frequency, the electroporated cells are also plated on chloramphenicol/LB plates at 30 deg C. From the 42 deg C plate, 1-5 colonies are picked into 1 ml of LB broth, serially diluted, and immediately plated at 30°ree;C on either 5% w/v sucrose or 5% sucrose+antibiotic plates. The 5% sucrose plates are replica plated to chloramphenicol plates at 30 deg C to test for loss of the replacement vector (cms). The gene replacement is confirmed by either PCR using primers flanking the targeted open reading frame or by genomic Southern's. Note from depositor: If sucrose plates do not select correctly, pKOV should be used in liquid selection on a plate reader with a LB + Cm control growth and a LB + Cm + sucrose growth. pKOV can sometimes still allow survival in the presence of sucrose, but significantly decreases fitness, so tracking the kinetic growth can be helpful. The sacB gene loses its efficacy,, sacB is toxic in E. coli even in the absence of sucrose.

Proper citation: RRID:Addgene_25769 Copy   

•   Source: Addgene