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Species: Homo sapiens
Genetic Insert: GB1-His-TET1-878-911-L900A
Vector Backbone Description: Backbone Marker:University of Leicester; Vector Backbone:pLeicester 91; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_124395 Copy
Species: Homo sapiens
Genetic Insert: FLAG-FullLength-TET1 with L897A/L900A mutations
Vector Backbone Description: Backbone Marker:University of Leicester; Vector Backbone:pLeicester 38; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_124397 Copy
Species: Homo sapiens
Genetic Insert: GB1-His-TET1-878-911 wildtype
Vector Backbone Description: Backbone Marker:University of Leicester; Vector Backbone:pLeicester 91; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_124390 Copy
Species: Homo sapiens
Genetic Insert: GB1-His-SAP25-126-186
Vector Backbone Description: Backbone Marker:University of Leicester; Vector Backbone:pLeicester 91; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_124387 Copy
Species: Homo sapiens
Genetic Insert: PSPC1
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:5446; Vector Backbone:pCDNA3; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
References:
Comments: For wildtype PSPC1 see https://www.addgene.org/101764/
Proper citation: RRID:Addgene_124373 Copy
Species:
Genetic Insert: Tornado-BroccoliSamSensor#1
Vector Backbone Description: Vector Backbone:pAV; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_124363 Copy
Species: Homo sapiens
Genetic Insert: Gal4-TET1-878-911 wildtype
Vector Backbone Description: Backbone Marker:University of Leicester; Vector Backbone:pLeicester 84; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_124400 Copy
Species: Synthetic
Genetic Insert: empty backbone (control)
Vector Backbone Description: Vector Backbone:pLG166; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_124368 Copy
Species: Homo sapiens
Genetic Insert: Gal4-TET1-878-911(I894A mutation)
Vector Backbone Description: Backbone Marker:University of Leicester; Vector Backbone:pLeicester 84; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_124404 Copy
Species: Homo sapiens
Genetic Insert: Gal4-TET1-878-911(T898A mutation)
Vector Backbone Description: Backbone Marker:University of Leicester; Vector Backbone:pLeicester 84; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_124403 Copy
Species: Synthetic
Genetic Insert: SpyDock
Vector Backbone Description: Backbone Marker:Thermo Fisher Scientific; Backbone Size:3641; Vector Backbone:pDEST14; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_124618 Copy
Species: Homo sapiens
Genetic Insert: PU.1 ETS domain (residues 167-272)
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:5428; Vector Backbone:pcDNA3.1; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_124616 Copy
Species:
Genetic Insert: TCF/LEF binding sites
Vector Backbone Description: Backbone Marker:Clontech; Backbone Size:4871; Vector Backbone:pTA-Luc; Vector Types:Luciferase; Bacterial Resistance:Ampicillin
References:
Comments: This is a luciferase reporter of beta-catenin-mediated transcriptional activation. In HEK cells, maximal activation of this reporter is ~100-fold (activation by Wnt) up to ~1,000-fold (activation by phosphorylation mutants of beta-catenin). The appropriate control plasmid is clone M51, Super8XFOPflash, which has mutant TCF/LEF binding sites.
This construct was made by Ajamete Kaykas in the Moon lab. The backbone is the pTA-Luc vector of Clontech, which provides a minimal TA viral promoter driving expression of the firefly luciferase gene (see company publications for details). 7 TCF/LEF binding sites were cloned into the Mlu1 site of this vector (7 copies of: AGATCAAAGGgggta, with TCF/LEF binding site in CAP letters, and a spacer in lower case, separating each copy of the TCF/LEF site).
Note: This plasmid was published as M50 Super 8x TOPFlash, but the plasmid actually contains 7 TCF/LEF sites.
Proper citation: RRID:Addgene_12456 Copy
Species: Homo sapiens
Genetic Insert: NPM1 Homology Arms with linker-mTagRFP-T
Vector Backbone Description: Vector Backbone:pUC57; Vector Types:Mammalian Expression, CRISPR, Other, Donor Template; Bacterial Resistance:Ampicillin
References:
Comments: This plasmid has been used with locus-specific CRISPR/Cas9 to add a mTagRFP-T tag to the C-terminus of human NPM1 in WTC human induced pluripotent stem cells by the Allen Institute for Cell Science. Linker (AA) sequence: KPNSAVDGTAGPGSIAT. 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_124608 Copy
Species: Mus musculus
Genetic Insert: UBP43
Vector Backbone Description: Backbone Marker:Amersham; Backbone Size:5000; Vector Backbone:pGEX-4T3; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_12458 Copy
Species: Homo sapiens
Genetic Insert: 14-3-3 sigma
Vector Backbone Description: Backbone Size:4679; Vector Backbone:pFlag-CMV2; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_12453 Copy
Species: Homo sapiens
Genetic Insert: EFP
Vector Backbone Description: Backbone Size:5400; Vector Backbone:pcDNA3.0; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
References:
Comments: Digestion by KpnI and XhoI should give a 5.4 kb fragment and a 2.4 kb fragment.
Compared to GenBank ID NP_005073.2, the EFP insert in this plasmid appears to contain a P385L mutation, whereas when compared to GenBank ID AB384815.1 the EFP insert appears to contain G89V and A185T mutations. These apparent mutations may represent valid polymorphisms and reflect the sequence of the original human cDNA clone. The plasmid is exactly as described in the associated publication.
Proper citation: RRID:Addgene_12452 Copy
Species: Mus musculus
Genetic Insert: UBP43
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:5400; Vector Backbone:pcDNA3.1; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_12454 Copy
Species: Mus musculus
Genetic Insert: Soma-localized ChrimsonR
Vector Backbone Description: Backbone Marker:Genscript; Backbone Size:3000; Vector Backbone:pcDNA; Vector Types:Mammalian Expression, AAV; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_124603 Copy
Species: Other
Genetic Insert: Protein A and hyperactive Tn5 transposase (Tnp) fusion protein
Vector Backbone Description: Backbone Size:8079; Vector Backbone:pTXB1-Tn5; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
References:
Comments: Please note that verifying this plasmid by sequencing or digest is challenging due to the presence of the helper plasmid, as multiple bands or mispriming is likely to occur. See Addgene's sequencing results to compare sequences.
This plasmid has been found to be somewhat unstable and prone to concatenation. Concatenation often does not impact plasmid function, but can reduce transformation or transfection efficiencies. If you have trouble isolating the monomeric version of this plasmid, you might consider linearizing, gel extracting, re-ligating, and transforming the plasmid.
Proper citation: RRID:Addgene_124601 Copy
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