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Genetic Insert: membrane bound form of EGFP
Vector Backbone Description: Backbone Size:6122; Vector Backbone:pCAGEN; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:17209010
Comments: mGFP is a membrane-bound form of GFP containing a palmitoylation sequence of GAP43 at its N-terminus.
The mGFP insert contains V163A and S175G compared to EGFP. These mutations are not known to affect plasmid function. The Cepko lab and several other Addgene users have used this plasmid successfully in experiments.
Addgene's NGS finds an IS4-like element (ISVsa5 family transposase) in the backbone that does not affect the overall plasmid function.
Proper citation: RRID:Addgene_14757 Copy
Species: Homo sapiens
Genetic Insert: GSK3 beta K85A
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:5500; Vector Backbone:pcDNA3; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:8382613
Comments: Nru I site is added to the K->A mutation
Proper citation: RRID:Addgene_14755 Copy
Species: Saccharomyces cerevisiae
Genetic Insert: Flpe-ERT2 fusion protein
Vector Backbone Description: Backbone Size:4779; Vector Backbone:pCAGEN; Vector Types:Mammalian Expression, Flp/FRT; Bacterial Resistance:Ampicillin
Defining Citation: PMID:17209010
Comments: Kozak consensus sequence was added before the start ATG. Internal EcoRI site was destroyed by introducing a silent mutation.
Proper citation: RRID:Addgene_14756 Copy
Species: Homo sapiens
Genetic Insert: GSK3 beta S9A
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:5500; Vector Backbone:pcDNA3; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:7980435
Comments: S2A in the sequence does not affect function, used to introduce restriction enzyme site
Proper citation: RRID:Addgene_14754 Copy
Species: N. pharaonis
Genetic Insert: Mammalian codon-optimized halorhodopsin, from Natronomas pharaonis (N. pharaonis), fused with GFP at the C-terminus
Vector Backbone Description: Backbone Marker:Pavel Osten; Backbone Size:9500; Vector Backbone:FCK(1.3)GW; Vector Types:Mammalian Expression, Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:17375185
Comments: PLEASE CONTACT ED BOYDEN ([email protected]) FOR DETAILS ON THIS
REAGENT AND FURTHER REAGENTS IN THIS LINE.
Proper citation: RRID:Addgene_14750 Copy
Species: Homo sapiens
Genetic Insert: N-ras
Vector Backbone Description: Backbone Size:7500; Vector Backbone:pCGN; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:11305105
Proper citation: RRID:Addgene_14725 Copy
Species: Homo sapiens
Genetic Insert: H-ras
Vector Backbone Description: Backbone Size:7500; Vector Backbone:pCGN; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:11305105
Comments: GTPase deficient (dominant active) mutant: GAP insensitive, increased GDP/GTP exchange, and chronically GTP bound; transforming.
Proper citation: RRID:Addgene_14720 Copy
Vector Backbone Description: Backbone Size:6794; Vector Types:Worm Expression; Bacterial Resistance:Ampicillin
Comments: See Fire Lab Vector Kit Documentation 1995. Fire Lab Miniprep Number pPD93.97, Ligation number NONE.
Proper citation: RRID:Addgene_1476 Copy
Vector Backbone Description: Backbone Size:4699; Vector Types:Worm Expression; Bacterial Resistance:Ampicillin
Comments: See Fire Lab Vector Kit Documentation 1995. Fire Lab Miniprep Number pPD94.81, Ligation number NONE.
Proper citation: RRID:Addgene_1477 Copy
Species: Homo sapiens
Genetic Insert: R-ras
Vector Backbone Description: Backbone Size:7500; Vector Backbone:pCGN; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:11305105
Proper citation: RRID:Addgene_14730 Copy
Species: Rattus norvegicus
Genetic Insert: MRF4
Vector Backbone Description: Backbone Size:5300; Vector Backbone:EMSV; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:10733231
Proper citation: RRID:Addgene_14713 Copy
Species: Homo sapiens
Genetic Insert: DGCR8 shRNA
Vector Backbone Description: Backbone Size:7700; Vector Backbone:pSicoR PGK puro; Vector Types:Mammalian Expression, Lentiviral, RNAi, Cre/Lox; Bacterial Resistance:Ampicillin
Defining Citation: PMID:17401365
Comments: shRNA sequence: TGGGGTGAGA GTGCTGATAA TTCAAGAGAT TATCAGCACT CTCACCCCTT TTTTC
Proper citation: RRID:Addgene_14769 Copy
Species: Homo sapiens
Genetic Insert: Drosha shRNA
Vector Backbone Description: Backbone Size:7700; Vector Backbone:pSicoR PGK puro; Vector Types:Mammalian Expression, Lentiviral, RNAi, Cre/Lox; Bacterial Resistance:Ampicillin
Defining Citation: PMID:17401365
Comments: shRNA sequence: TGAAGCTCGA TGAAGATTTA TTCAAGAGAT AAATCTTCAT CGAGCTTCTT TTTTC
Proper citation: RRID:Addgene_14766 Copy
Species: Homo sapiens
Genetic Insert: Drosha shRNA
Vector Backbone Description: Backbone Size:7700; Vector Backbone:pSicoR PGK puro; Vector Types:Mammalian Expression, Lentiviral, RNAi, Cre/Lox; Bacterial Resistance:Ampicillin
Defining Citation: PMID:17401365
Comments: shRNA sequence: TGAAGCTCTT TGGTGAATAA TTCAAGAGAT TATTCACCAA AGAGCTTCTT TTTTC
Proper citation: RRID:Addgene_14767 Copy
Genetic Insert: EGFPd2
Vector Backbone Description: Backbone Size:4779; Vector Backbone:pCAGEN; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:17209010
Proper citation: RRID:Addgene_14760 Copy
Species: Mus musculus
Genetic Insert: Hmga2 wt
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:5500; Vector Backbone:pcDNA3.1; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:17322030
Comments: The 3' UTR of the mouse Hmga2 cDNA (BC052158) was subcloned into pCR2.1-TOPO (Invitrogen) for site-directed mutagenesis. To disrupt each let-7 complementary site, the nucleotides that paired to nucleotides 3 and 5 of the miRNA were substituted (see Author's map), using the Quikchange site-directed mutagenesis kit (Stratagene). The UTR fragment was then cloned back into the vector containing the Hmga2 cDNA (pCMV•SPORT6.1). To construct the mammalian expression vectors, Hmga2 inserts were excised from the pCMV•SPORT6.1 constructs using EcoRI restriction sites and cloned into pcDNA3.1 (Invitrogen).
Proper citation: RRID:Addgene_14789 Copy
Species: Mus musculus
Genetic Insert: let-7g
Vector Backbone Description: Backbone Marker:BD Biosciences; Backbone Size:6500; Vector Backbone:MSCV-neo; Vector Types:Mammalian Expression, Retroviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:17401365
Comments: miRNA of the following mature sequence, with approximately 500 basepairs of flanking sequence, were cloned into MSCV-neo: let-7g: 5’-UGAGG UAGUA GUUUG UACAG U-3’.
Proper citation: RRID:Addgene_14784 Copy
Species: Mus musculus
Genetic Insert: Luc-wt
Vector Backbone Description: Backbone Marker:Available from Addgene (#12179); Backbone Size:4000; Vector Backbone:pIS1; Vector Types:Mammalian Expression, Luciferase; Bacterial Resistance:Ampicillin
Defining Citation: PMID:17322030
Comments: The 3' UTR of the mouse Hmga2 cDNA (BC052158) was subcloned into pCR2.1-TOPO (Invitrogen) for site-directed mutagenesis. To disrupt each let-7 complementary site, the nucleotides that paired to nucleotides 3 and 5 of the miRNA were substituted (see Author's map), using the Quikchange site-directed mutagenesis kit (Stratagene). To construct the Renilla luciferase reporters, wild-type and mutant Hmga2 3' UTRs were amplified (PCR primers, 5'-GCGTCTCGAGGGGCGCCGACATTC and 5'GGCGCGGCCGCAGTCAGAGGGCACAC) and cloned into the XbaI and NotI sites of pIS1.
Proper citation: RRID:Addgene_14785 Copy
Species: Homo sapiens
Genetic Insert: Smad3 NL
Vector Backbone Description: Backbone Marker:Genentech; Backbone Size:4700; Vector Backbone:pRK5; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:9094310
Comments: See Author's map for cloning details.
Proper citation: RRID:Addgene_14828 Copy
Species: Homo sapiens
Genetic Insert: Retinoblastoma Binding Protein 2
Vector Backbone Description: Backbone Size:5446; Vector Backbone:pcDNA3; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:17320163
Comments: The plasmid encodes isoform 2.
Proper citation: RRID:Addgene_14799 Copy
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