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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
Vector Backbone Description: Backbone Size:3586; Vector Backbone:pBTB-3; Vector Types:Bacterial Expression; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:16496398
Comments: Addgene's QC sequence shows a 4bp gap with the depositor's provided sequence. The lab does not believe this affects the plasmid activity.
Proper citation: RRID:Addgene_22819 Copy
Species: Synechocystis/E.coli fusion gene
Genetic Insert: pCph8
Vector Backbone Description: Backbone Marker:Clontech; Backbone Size:2021; Vector Backbone:pPROTet; Vector Types:Bacterial Expression, Synthetic Biology; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:16306980
Comments: This plasmid was also used in the 2009 Cell paper: "A synthetic genetic edge detection program." PMID:19563759
Proper citation: RRID:Addgene_22869 Copy
Vector Backbone Description: Backbone Size:4831; Vector Backbone:pBMTB-3; Vector Types:Bacterial Expression; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:16496398
Comments: Addgene's QC sequence shows a 4bp gap with the depositor's provided sequence. The lab does not believe this affects the plasmid activity.
Proper citation: RRID:Addgene_22826 Copy
Vector Backbone Description: Backbone Size:3649; Vector Backbone:pBMT-3; Vector Types:Bacterial Expression; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:16496398
Proper citation: RRID:Addgene_22838 Copy
Species: Mus musculus
Genetic Insert: DHFR
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:4000; Vector Backbone:pBAD/HisA & pBThybrid; Vector Types:Bacterial Expression; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:19165721
Proper citation: RRID:Addgene_23011 Copy
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
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
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
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
Species: Homo sapiens
Genetic Insert: p21 promoter deletion mutant
Vector Backbone Description: Backbone Marker:Stratagene; Backbone Size:5600; Vector Backbone:pBlueScript II KS(+); Vector Types:Mammalian Expression, Luciferase; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:8988173
Proper citation: RRID:Addgene_21728 Copy
Species: Homo sapiens
Genetic Insert: p21 promoter deletion mutant
Vector Backbone Description: Backbone Marker:Stratagene; Backbone Size:5600; Vector Backbone:pBlueScript II KS(+); Vector Types:Mammalian Expression, Luciferase; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:8988173
Proper citation: RRID:Addgene_21730 Copy
Species: L. pneumophila
Genetic Insert: fabI
Vector Backbone Description: Backbone Size:9869; Vector Backbone:pXDC61; Vector Types:Bacterial Expression; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:19578436
Proper citation: RRID:Addgene_21842 Copy
Species: Mesoplasma florum
Genetic Insert: Lon
Vector Backbone Description: Backbone Marker:Beckwith Lab; Backbone Size:5352; Vector Backbone:pBAD33; Vector Types:Bacterial Expression; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:18852454
Comments: A detailed protocol for purification of mf-lon can be found in the associated paper.
Proper citation: RRID:Addgene_21867 Copy
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
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