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Species: Homo sapiens
Genetic Insert: miniTurbo-V5-RNASEL
Vector Backbone Description: Backbone Marker:PEL; Backbone Size:9019; Vector Backbone:pDest663; Vector Types:Mammalian Expression, Lentiviral, Gateway Destination; Bacterial Resistance:Ampicillin
Comments: These plasmids were generated as part of the Illuminating the Druggable Genome (IDG) program sponsored by the NIH Common Fund. The goal of this program is to identify, gather, and distribute information and resources for proteins that currently are not well-studied yet belong to commonly drug-targeted protein families: protein kinases, non-olfactory G-protein coupled receptors (GPCRs), and ion channels. The IDG program is designed to develop fundamental research tools for understudied proteins, elucidate their function, and disseminate the IDG-related resources and data to the greater scientific community. These lentiviral gateway destination plasmids were generated, as part of the Kinase-Data and Resource Generating Center (DRGC), using the single fragment or multisite gateway cloning technology. They were used for generating proteomics-based protein-interaction and protein-proximity networks that can be accessed through the kinase-DRGC website https://darkkinome.org/. Each plasmid has either an N- or C-terminal fusion protein (Flag/ V5/V5-miniTurbo/V5-TurboID/miniTurbo-V5/TurboID-V5) tagged understudied kinase driven under either a CMV or Ubiquitin (UBC) promoter. All inserts in the pENTR plasmids used to generate the deposited destination plasmids were end-sequenced and insert size validated prior to multi-site gateway cloning. The combined insert size of the single fragment or three fragment destination plasmids were confirmed by restriction digestion (BsrGI, and EcoRV for pDest667; BsrGI, EcoRV, and BstZ17I for pDest663; and BsrGI for pHAGE) and all plasmids were partially sequenced to ensure in-frame ORFs and fusion tags.
Proper citation: RRID:Addgene_170604 Copy
Species: Homo sapiens
Genetic Insert: miniTurbo-V5-RPS6KL1
Vector Backbone Description: Backbone Marker:PEL; Backbone Size:9019; Vector Backbone:pDest663; Vector Types:Mammalian Expression, Lentiviral, Gateway Destination; Bacterial Resistance:Ampicillin
Comments: These plasmids were generated as part of the Illuminating the Druggable Genome (IDG) program sponsored by the NIH Common Fund. The goal of this program is to identify, gather, and distribute information and resources for proteins that currently are not well-studied yet belong to commonly drug-targeted protein families: protein kinases, non-olfactory G-protein coupled receptors (GPCRs), and ion channels. The IDG program is designed to develop fundamental research tools for understudied proteins, elucidate their function, and disseminate the IDG-related resources and data to the greater scientific community. These lentiviral gateway destination plasmids were generated, as part of the Kinase-Data and Resource Generating Center (DRGC), using the single fragment or multisite gateway cloning technology. They were used for generating proteomics-based protein-interaction and protein-proximity networks that can be accessed through the kinase-DRGC website https://darkkinome.org/. Each plasmid has either an N- or C-terminal fusion protein (Flag/ V5/V5-miniTurbo/V5-TurboID/miniTurbo-V5/TurboID-V5) tagged understudied kinase driven under either a CMV or Ubiquitin (UBC) promoter. All inserts in the pENTR plasmids used to generate the deposited destination plasmids were end-sequenced and insert size validated prior to multi-site gateway cloning. The combined insert size of the single fragment or three fragment destination plasmids were confirmed by restriction digestion (BsrGI, and EcoRV for pDest667; BsrGI, EcoRV, and BstZ17I for pDest663; and BsrGI for pHAGE) and all plasmids were partially sequenced to ensure in-frame ORFs and fusion tags.
Proper citation: RRID:Addgene_170605 Copy
Species: Homo sapiens
Genetic Insert: CPTP
Vector Backbone Description: Vector Backbone:pFLAG; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:23863933
Proper citation: RRID:Addgene_170744 Copy
Species: Homo sapiens
Genetic Insert: CPTP
Vector Backbone Description: Vector Backbone:pFLAG; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:21625605
Proper citation: RRID:Addgene_170742 Copy
Species: Homo sapiens
Genetic Insert: GLTP
Vector Backbone Description: Vector Backbone:pFLAG; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:21625605
Proper citation: RRID:Addgene_170740 Copy
Species: Homo sapiens
Genetic Insert: LAMP1
Vector Backbone Description: Backbone Marker:Addgene (Plasmid #14883); Backbone Size:9955; Vector Backbone:FUGW; Vector Types:Mammalian Expression, Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:34102054
Comments: Chin et al. ACS Sens. 2021 Jun 8. doi: 10.1021/acssensors.0c02318.
Pubmed: https://pubmed.ncbi.nlm.nih.gov/34102054/
Article: https://pubs.acs.org/doi/10.1021/acssensors.0c02318
Proper citation: RRID:Addgene_170774 Copy
Species: Homo sapiens
Genetic Insert: LAMP1
Vector Backbone Description: Backbone Marker:Addgene Plasmid #19319; Backbone Size:8083; Vector Backbone:pLJM1-EGFP; Vector Types:Mammalian Expression, Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:34102054
Comments: Chin et al. ACS Sens. 2021 Jun 8. doi: 10.1021/acssensors.0c02318.
Pubmed: https://pubmed.ncbi.nlm.nih.gov/34102054/
Article: https://pubs.acs.org/doi/10.1021/acssensors.0c02318
Proper citation: RRID:Addgene_170775 Copy
Species: Homo sapiens
Genetic Insert: PCNA Homology Arms with mEGFP-linker
Vector Backbone Description: Vector Backbone:pUC57; Vector Types:Mammalian Expression, CRISPR, Donor Template; Bacterial Resistance:Ampicillin
Comments: This plasmid has been used with locus-specific CRISPR/Cas9 to add a mEGFP tag to the N-terminus of human PCNA in WTC-11 human induced pluripotent stem cells by the Allen Institute for Cell Science. Linker (AA) sequence: GEGQGQGQGPGRGYAYRS. After protein tagging using this donor template plasmid and CRISPR/Cas9 reagents, transfected cells may exhibit varying intensity levels of fluorescence, likely due to editing precision. To obtain cells of uniform intensity levels, see our protocol for fluorescence-assisted cell sorting and subcloning of transfected cells (https://www.allencell.org/instructional-videos-and-tutorials-for-cell-methods.html). Further, we recommend PCR-based assays for identifying precisely edited clones as previously described (https://www.molbiolcell.org/doi/abs/10.1091/mbc.e17-03-0209). For more information on the entire plasmid collection, please see https://www.addgene.org/allen-institute-cell-science/ .
Proper citation: RRID:Addgene_170818 Copy
Species: Homo sapiens
Genetic Insert: ERGIC-53
Vector Backbone Description: Backbone Marker:Clontech; Backbone Size:4026; Vector Backbone:pDendra2-C; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:35051356
Comments: Please see the Genbank file under Resource Information for an annotated sequence file from the depositing laboratory.
Proper citation: RRID:Addgene_170717 Copy
Species: Homo sapiens
Genetic Insert: STAT5A
Vector Backbone Description: Vector Backbone:pTRIPZ; Vector Types:Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:34263728
Comments: A constitutively active form of STAT5A was cloned into the lentiviral expression vector.
Proper citation: RRID:Addgene_170662 Copy
Species: Homo sapiens
Genetic Insert: W96A-GLTP
Vector Backbone Description: Vector Backbone:pET30 Xa/LIC; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:15329726
Proper citation: RRID:Addgene_170734 Copy
Species: Homo sapiens
Genetic Insert: GLTP
Vector Backbone Description: Vector Backbone:pSUMO; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:30206120
Proper citation: RRID:Addgene_170732 Copy
Species: Homo sapiens
Genetic Insert: GLTP
Vector Backbone Description: Vector Backbone:pET30 Xa/LIC; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:15329726
Proper citation: RRID:Addgene_170733 Copy
Species: Homo sapiens
Genetic Insert: K60A-CPTP
Vector Backbone Description: Vector Backbone:pSUMO; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:23863933
Proper citation: RRID:Addgene_170736 Copy
Species: Homo sapiens
Genetic Insert: E. coli ornithine transcarbamylase (OTC), fused to human ubiquitin
Vector Backbone Description: Backbone Marker:Novagen (+ 6xHis-TEV in-house modification); Backbone Size:3810; Vector Backbone:pRSFDuet1-6xHis-TEV; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:33719392
Proper citation: RRID:Addgene_170729 Copy
Species: Homo sapiens
Genetic Insert: Synaptotagmin-like 5
Vector Backbone Description: Backbone Marker:Clonetech; Vector Backbone:pEGFP-C1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:34172739
Proper citation: RRID:Addgene_170868 Copy
Species: Homo sapiens
Genetic Insert: Human GABAA receptor alpha1 subunit
Vector Backbone Description: Vector Backbone:pIRES2-EGFP; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:30481215
Proper citation: RRID:Addgene_170820 Copy
Species: Homo sapiens
Genetic Insert: Human GABAA receptor alpha1 subunit carrying A322D mutation
Vector Backbone Description: Vector Backbone:pIRES2-EGFP; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:30481215
Proper citation: RRID:Addgene_170821 Copy
Species: Homo sapiens
Genetic Insert: RE8
Vector Backbone Description: Vector Backbone:pRF TpC; Vector Types:Mammalian Expression, Luciferase; Bacterial Resistance:Ampicillin
Defining Citation: PMID:33852895
Proper citation: RRID:Addgene_171047 Copy
Species: Homo sapiens
Genetic Insert: RE7
Vector Backbone Description: Vector Backbone:pRF TpC; Vector Types:Luciferase; Bacterial Resistance:Ampicillin
Defining Citation: PMID:33852895
Proper citation: RRID:Addgene_171046 Copy
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