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Vector Backbone Description: Backbone Size:6447; Vector Backbone:JDS80; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Comments: Backbone contains SNI/A 0.5 Domain
TAL N-terminal: ∆152 (Miller et al. 2011)
TAL C-terminus: +63 (Miller et al. 2011)
FOKI: KKR (+) heterodimer (Doyon et al. 2011)
Proper citation: RRID:Addgene_40316 Copy
Species: Saccharomyces cerevisiae
Genetic Insert: LEU2
Vector Backbone Description: Backbone Marker:ATCC, Number: 77142; Backbone Size:4967; Vector Backbone:pRS313; Vector Types:Yeast Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:23222782
Proper citation: RRID:Addgene_40276 Copy
Species: Homo sapiens
Genetic Insert: erbB1
Vector Backbone Description: Backbone Marker:Clontech; Vector Backbone:pEGFP-N3; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:17956594
Proper citation: RRID:Addgene_40267 Copy
Species: Homo sapiens
Genetic Insert: erbB1
Vector Backbone Description: Backbone Marker:Clontech; Vector Backbone:pEYFP-N3; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:17956594
Proper citation: RRID:Addgene_40266 Copy
Species: Homo sapiens
Genetic Insert: HTT partial exon 1 Q23
Vector Backbone Description: Backbone Marker:Roche; Backbone Size:5450; Vector Backbone:pHM6; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:10528852
Proper citation: RRID:Addgene_40263 Copy
Species: Homo sapiens
Genetic Insert: HTT partial exon 1 Q23
Vector Backbone Description: Backbone Marker:Clontech; Backbone Size:4700; Vector Backbone:pEGFP-C1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:10528852
Proper citation: RRID:Addgene_40261 Copy
Species: Synthetic
Genetic Insert: Clover GFP
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:5410; Vector Backbone:pcDNA3; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:22961245
Comments: Clover is the brightest green fluorescent protein (GFP) characterized to date, with robust performance in protein fusions. Also, it is useful as a FRET donor to mRuby2 in a FRET pair that offers bright fluorescence, dynamic range, and photostability while limiting emissions overlap.
Proper citation: RRID:Addgene_40259 Copy
Species: Synthetic
Genetic Insert: VSFP-CR
Vector Backbone Description: Backbone Marker:Invitrogen/Dr. Paulmurugan Ramasamy; Backbone Size:6107; Vector Backbone:pcDNA3.1/Puro-CAG; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:22961245
Comments: Voltage sensor domain of Ci-VSP fused to a pair of fluorescent proteins consisting of a Clover GFP and mRuby2 RFP.
Proper citation: RRID:Addgene_40257 Copy
Species: Synthetic
Genetic Insert: AKAR2
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:5489; Vector Backbone:pcDNA3; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:22961245
Comments: ECFP was replaced with GFP Clover and Citrine was replaced with RFP mRuby2.
Note: there is a small sequence insertion downstream of the insert that adds restriction sites. This is not present in the full plasmids sequence, so please refer to Addgene's QC sequence with the BGH-Rev primer for additional information.
Proper citation: RRID:Addgene_40255 Copy
Species: Homo sapiens
Genetic Insert: drebrin E2
Vector Backbone Description: Backbone Marker:EMD Biosciences; Backbone Size:5420; Vector Backbone:pETDuet-1; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:18806788
Proper citation: RRID:Addgene_40362 Copy
Species: Synechococcus elongatus PCC 7942
Genetic Insert: Km Resistance, neutral site II integration platform for Synechococcus
Vector Backbone Description: Vector Backbone:1572; Vector Types:Cyanobacteria cloning vector; Bacterial Resistance:Ampicillin and Kanamycin
Defining Citation: PMID:10812624
Comments: NSII vector
cloning sites: NheI, XbaI, StuI, SalI, and EcoRV.
Km cassette into Klenow blunted BglII site of pAM1572. XhoI digest gives 0.7kb band. NSII vector; unique cloning sites NheI, XbaI, StuI, SalI, EcoRV.
Proper citation: RRID:Addgene_40240 Copy
Species: Rattus norvegicus
Genetic Insert: GCaMP6s
Vector Backbone Description: Backbone Marker:Clontech; Backbone Size:3955; Vector Backbone:pEGFP-N1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:23868258
Proper citation: RRID:Addgene_40753 Copy
Species: Homo sapiens
Genetic Insert: POU5F1_S234A_T235A
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:10712; Vector Backbone:pCEP4; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:22474382
Proper citation: RRID:Addgene_40630 Copy
Species: Arabidopsis thaliana
Genetic Insert: CRY2
Vector Backbone Description: Backbone Size:5414; Vector Backbone:p414TEF; Vector Types:Yeast Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:22922268
Proper citation: RRID:Addgene_40593 Copy
Species: Homo sapiens
Genetic Insert: Cre recombinase
Vector Backbone Description: Backbone Marker:Michael Brenner (PMID: 7952266); Backbone Size:5500; Vector Backbone:pgfa2-lac2; Vector Types:Mammalian Expression, Mouse Targeting, Cre/Lox; Bacterial Resistance:Ampicillin
Defining Citation: PMID:11668683
Comments: This plasmid encodes a transgene to express the Cre recombinase in astrocytes of transgenic mice. A DNA fragment encoding the Cre recombinase was inserted into an expression cassette containing the gfa2 promoter (Besnard et al., 1991), a 2.2-kb 5' flanking region from the human GFAP (hGFAP) gene. The nuclear targeting signal from the SV40 large T antigen was inserted at the amino terminal end of the coding region, because it reportedly increases the efficiency of DNA excision by the recombinase (Gu et al., 1993). A heterologous intron and polyadenylation signal were provided by sequences from the mouse protamine 1 gene at the 3' end.
The hGFAP-cre transgene was constructed by excising the lacZ coding region from pgfa2-lac2 (Brenner et al., 1994) by BamHI digestion, and replacing it by blunt end ligation with a nuclear-targeted Cre coding sequence. The latter was obtained as a 1.1-kb SalI-MluI fragment from plasmid pTZ19RCreNLS, which was kindly provided by Drs. Raul Torres and Klaus Rajewsky. Both junctions were verified by DNA sequencing.
Proper citation: RRID:Addgene_40591 Copy
Species: Mus musculus
Genetic Insert: Syndecan-1 shRNA(UTR)
Vector Backbone Description: Backbone Size:10130; Vector Backbone:FUGW-H1; Vector Types:Lentiviral, RNAi; Bacterial Resistance:Ampicillin
Defining Citation: PMID:22936997
Comments: This shRNA plasmid was generated by inserting the hairpin oligonucleotides into the FUGW-H1
lentiviral construct.
Proper citation: RRID:Addgene_40623 Copy
Species: Homo sapiens
Genetic Insert: Wnt reporter sequences
Vector Backbone Description: Backbone Marker:addgene; Vector Backbone:pSicoR PGK Puro; Vector Types:Mammalian Expression, Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:22177620
Comments: This lentiviral Wnt reporter has 7 repeats of Tcf/Lef binding sites, which was cloned by Xiaojun (Lance) Lian.
Methods for Plasmid construction, lentiviral production and infection of hESCs:
The 7 TCF/LEF binding site sequences were obtained by direct PCR of plasmid M50 (Addgene plasmid 12456) and the GreenFire (GF) sequences were obtained by direct PCR of the pGreenFire1-mCMV Plasmid (System Bioscience TR010PA-1). These two sequences were cloned into pSicoR PGK puro (Addgene plasmid 11586) digested by Xba I (NEB) and Xho I (NEB). The constructed Wnt reporter plasmid was named 7TGFP and verified by sequencing. This 7TGFP vector was cotransfected with the helper plasmids psPAX2 and pMD2.G (Addgene plasmids 12260 and 12259) into HEK-293TN cells (System Biosciences) for virus production. Virus-containing medium was collected at 48 and 72 hours after transfection and used for infection of hESCs in the presence of 6μg/mL polybrene (Sigma). Transduced cells were cultured in mTeSR1 on Matrigel for three days and then clonally isolated in mTeSR1 with 1 μg/mL puromycin.
Proper citation: RRID:Addgene_40588 Copy
Species: Homo sapiens
Genetic Insert: TCF7
Vector Backbone Description: Backbone Size:5400; Vector Backbone:pcDNA3; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:22949634
Proper citation: RRID:Addgene_40620 Copy
Species: Synthetic
Genetic Insert: Leucine Zipper prime - C super positive Green Fluorescent Protein
Vector Backbone Description: Backbone Size:4496; Vector Backbone:pMRBAD; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:22692102
Proper citation: RRID:Addgene_40730 Copy
Species: Mus musculus
Genetic Insert: AMPK-γ1
Vector Backbone Description: Backbone Size:4300; Vector Backbone:pCMV-Tag4A; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:17012231
Comments: AMPK-γ1 insert has Kozak sequence embedded at 5'-terminus and a stop codon at the 3'-terminus. The FLAG of the pCMV-Tag4a backbone is out of frame.
Proper citation: RRID:Addgene_40605 Copy
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