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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
Vector Backbone Description: Backbone Size:3691; Vector Backbone:pBTBXh-3; Vector Types:Bacterial Expression; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:20148414
Comments: There is an extra G in the middle of the alignment between Addgene's quality control sequence and the author's sequence. This discrepancy is in a non-coding region and does affect function.
Proper citation: RRID:Addgene_26079 Copy
Vector Backbone Description: Backbone Size:4922; Vector Backbone:pBMTBX-3; Vector Types:Bacterial Expression; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:20148414
Proper citation: RRID:Addgene_26074 Copy
Species: Aequorea victoria
Genetic Insert: destabilized GFP
Vector Backbone Description: Backbone Size:0; Vector Backbone:pLuxI-tet8 + pRKM-102; Vector Types:Bacterial Expression; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:15096621
Comments: A novel hybrid promoter luxPRcI-OR1 (wild-type luxPR promoter with a CI OR1 operator site inserted at the +1 transcription start) drives expression of GFP(LVA). The promoter was constructed by encoding OR1 on PCR primers and inserting it after luxPR
Proper citation: RRID:Addgene_26580 Copy
Species: E. coli
Genetic Insert: lacIq
Vector Backbone Description: Backbone Size:4945; Vector Backbone:N/A; Vector Types:Bacterial Expression; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:17565681
Comments: Protocol for preparing proteins with improved solubility by co-expressing with molecular chaperones in Escherichia coli.
de Marco A.
Nat Protoc. 2007;2(10):2632-9.
Proper citation: RRID:Addgene_27390 Copy
Genetic Insert: qtag22
Vector Backbone Description: Backbone Marker:N/A; Backbone Size:4522; Vector Backbone:pTCS-2M1-icl2; Vector Types:Bacterial Expression; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:21203517
Proper citation: RRID:Addgene_27335 Copy
Genetic Insert: qtag17
Vector Backbone Description: Backbone Marker:N/A; Backbone Size:4522; Vector Backbone:pTCS-2M1-icl2; Vector Types:Bacterial Expression; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:21203517
Proper citation: RRID:Addgene_27333 Copy
Genetic Insert: qtag20
Vector Backbone Description: Backbone Marker:N/A; Backbone Size:5740; Vector Backbone:pDE43-MCKq1; Vector Types:Gateway Cloning; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:21203517
Proper citation: RRID:Addgene_27334 Copy
Species: Synthetic
Genetic Insert: J306 scRNA
Vector Backbone Description: Backbone Marker:Jesse Zalatan; Backbone Size:1726; Vector Backbone:pCK669 (Addgene #192638); Vector Types:CRISPR, Synthetic Biology; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:36378874
Proper citation: RRID:Addgene_192641 Copy
Species: S. pyogenes (dCas9)
Genetic Insert: Sp.pCas9-dSpRY
Vector Backbone Description: Backbone Marker:Jesse Zalatan; Backbone Size:1726; Vector Backbone:pCD442 (Addgene #153024); Vector Types:CRISPR, Synthetic Biology; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:36378874
Proper citation: RRID:Addgene_192640 Copy
Species: S. pyogenes (dCas9)
Genetic Insert: Sp.pCas9-dCas9-NG
Vector Backbone Description: Backbone Marker:Jesse Zalatan; Backbone Size:1726; Vector Backbone:pCD442 (Addgene #153024); Vector Types:CRISPR, Synthetic Biology; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:36378874
Proper citation: RRID:Addgene_192637 Copy
Species: Arabidopsis thaliana
Genetic Insert: SulR
Vector Backbone Description: Backbone Marker:GoldenBraid kit from Diego Orzaez (Addgene kit # 1000000076 ), IBMCP, Valencia, Spain; Backbone Size:2106; Vector Backbone:pUPD2; Vector Types:Plant Expression, Synthetic Biology, CDS (part B3-B4-B5), GoldenBraid level 0; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:36609768
Proper citation: RRID:Addgene_186423 Copy
Species: Synthetic
Genetic Insert: synGR (Part 3)
Vector Backbone Description: Backbone Size:1658; Vector Backbone:pYKT001; Vector Types:Synthetic Biology; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:36367334
Proper citation: RRID:Addgene_194480 Copy
Species: Synthetic
Genetic Insert: tCYC1 (Part 4)
Vector Backbone Description: Backbone Size:1658; Vector Backbone:pYKT001; Vector Types:Synthetic Biology; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:36367334
Proper citation: RRID:Addgene_194492 Copy
Species: Synthetic
Genetic Insert: synDHBR (Part 3)
Vector Backbone Description: Backbone Size:1658; Vector Backbone:pYKT001; Vector Types:Synthetic Biology; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:36367334
Proper citation: RRID:Addgene_194478 Copy
Species: Synthetic
Genetic Insert: GB_SynP (A2) G1ab.3
Vector Backbone Description: Vector Backbone:pUPD2; Vector Types:Synthetic Biology; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:36044643
Proper citation: RRID:Addgene_193124 Copy
Species: Synthetic
Genetic Insert: 37-12 Op (Part 2)
Vector Backbone Description: Backbone Size:1658; Vector Backbone:pYKT001; Vector Types:Synthetic Biology; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:36367334
Proper citation: RRID:Addgene_194490 Copy
Species: Synthetic
Genetic Insert: Zif268 Op (Part 2)
Vector Backbone Description: Backbone Size:1658; Vector Backbone:pYKT001; Vector Types:Synthetic Biology; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:36367334
Proper citation: RRID:Addgene_194491 Copy
Species: Synthetic
Genetic Insert: GB_SynP (A2) G1ab.4
Vector Backbone Description: Vector Backbone:pUPD2; Vector Types:Synthetic Biology; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:36044643
Proper citation: RRID:Addgene_193126 Copy
https://dgrc.bio.indiana.edu/stock/1024724
Species: Drosophila melanogaster
Defining Citation: PMID:12176937
Comments: BDGP ESTs
Proper citation: RRID:DGRC_1024724 Copy
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The data sets included in this resource are published in Kravitz et al, 2012. Distinct roles for direct and indirect pathway striatal neurons in reinforcement (Nat. Neurosci).
