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Genetic Insert: Ras binding domain of Raf
Vector Backbone Description: Backbone Marker:Promega; Backbone Size:4000; Vector Backbone:pCI; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:16429133
Proper citation: RRID:Addgene_18664 Copy
Species: Homo sapiens
Genetic Insert: H-Ras
Vector Backbone Description: Backbone Marker:Promega; Backbone Size:4000; Vector Backbone:pCI; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:16429133
Proper citation: RRID:Addgene_18665 Copy
Vector Backbone Description: Backbone Marker:Novagen; Backbone Size:7934; Vector Backbone:pET19b; Vector Types:Bacterial Expression, Gateway Cloning; Bacterial Resistance:Chloramphenicol and Ampicillin
Defining Citation: PMID:18064608
Comments: pET19b Gateway-compatible Destination vector. Combine with an Entry clone containing your gene of interest with the Gateway LR reaction to generate an expression clone containing an N-terminal His tag. Note that the Addgene verified sequence detects a mutation in the enterokinase site D(V)DDK.
Proper citation: RRID:Addgene_18668 Copy
Vector Backbone Description: Backbone Size:7040; Vector Backbone:pEGFPC4; Vector Types:Mammalian Expression; Bacterial Resistance:Chloramphenicol and Kanamycin
Defining Citation: PMID:18064608
Comments: pEGFPC4 Gateway-compatible Destination vector. Combine with an Entry clone containing your gene of interest with the Gateway LR reaction to generate an expression clone containing an N-terminal EGFP tag.
Proper citation: RRID:Addgene_18671 Copy
Species: Bacteriophage T4
Genetic Insert: T4 Lysozyme
Vector Backbone Description: Backbone Size:5000; Vector Backbone:pHS1403; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Comments: The BamHI site is 27 bp from the ATG and the HindIII site is 115 bp from the stop.
Proper citation: RRID:Addgene_18640 Copy
Genetic Insert: scramble
Vector Backbone Description: Backbone Marker:Stewart SA, RNA 2003 Apr; 9(4):493-501.; Backbone Size:7032; Vector Backbone:pLKO.1; Vector Types:Mammalian Expression, Lentiviral, RNAi; Bacterial Resistance:Ampicillin
Defining Citation: PMID:15718470
Comments: shRNA for use as a negative control. Sequence of hairpin is:CCTAAGGTTAAGTCGCCCTCGCTCGAGCGAGGGCGACTTAACCTTAGG. For packaging, please use pCMV-dR8.2 dvpr (Addgene plasmid #8455) and pCMV-VSVG (Addgene plasmid #8454).
Please note that the 5' cloning site, AgeI, is typically destroyed during the shRNA cloning. Depending on the specific shRNA sequence, the site can occasionally be restored. AgeI is present in this plasmid.
Proper citation: RRID:Addgene_1864 Copy
Genetic Insert: Gateway cassette A
Vector Backbone Description: Backbone Size:4750; Vector Backbone:pMXs; Vector Types:Mammalian Expression, Retroviral; Bacterial Resistance:Chloramphenicol and Ampicillin
Defining Citation: PMID:16904174
Comments: The sequence of primer pMX-S1811 is GAC GGC ATC GCA GCT TGG ATA CAC.
Proper citation: RRID:Addgene_18656 Copy
Species: Homo sapiens
Genetic Insert: 8-oxoguanine DNA glycosylase
Vector Backbone Description: Backbone Size:5000; Vector Backbone:pMCV/myc/nuc; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:17018150
Proper citation: RRID:Addgene_18709 Copy
Vector Backbone Description: Backbone Size:0; Vector Backbone:MSCV Luciferase PGK Hygro; Vector Types:Mammalian Expression, Retroviral, Luciferase; Bacterial Resistance:Ampicillin
Proper citation: RRID:Addgene_18782 Copy
Species: Bacillus subtilis
Genetic Insert: TRAP
Vector Backbone Description: Backbone Size:4200; Vector Backbone:n/a; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:17023487
Comments: Mammalian expression vector for the expression of HA and His6 epitope-tagged TRAP. There is a single amino acid (glycine) insertion between translational initiator codon and second amino acid of TRAP (asparagine) to maintain Kozak consensus sequence. Also, there is a single amino acid difference at amino acid #60 which has an asparagine (N) as in Bacillus amyloliquefaciens rather than glutamic acid (E) as in Bacillus subtilis. Codons are a mixture of human, Bacillus amyloliquefaciens, and Bacillus subtilis.
Proper citation: RRID:Addgene_18785 Copy
Genetic Insert: TRAP binding site
Vector Backbone Description: Backbone Marker:Available at Addgene (#18673); Backbone Size:6270; Vector Backbone:pYIC; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:17023487
Comments: An oligonucleotide with a 55-nt TRAP-binding sequence, TBS, was inserted into the 5'UTR of pYIC DNA to generate a bicistronic reporter gene with TBS: The TBS was inserted in the SacI site for 5-UTR (pTBS/5Y-YIC; 45 nt downstream of transcription start site and 9 nt upstream of YFP translational start site).
Proper citation: RRID:Addgene_18786 Copy
Vector Backbone Description: Backbone Marker:Sigma; Backbone Size:6932; Vector Backbone:pFlag-CMV-2; Vector Types:Mammalian Expression; Bacterial Resistance:Chloramphenicol and Ampicillin
Defining Citation: PMID:18064608
Comments: pFLAG-CMV-2 Gateway-compatible Destination vector. Combine with an Entry clone containing your gene of interest with the Gateway LR reaction to generate an expression clone containing an N-terminal FLAG tag.
Proper citation: RRID:Addgene_18700 Copy
Species: Mus musculus
Genetic Insert: epidermal growth factor receptor
Vector Backbone Description: Backbone Marker:Sigma; Backbone Size:6280; Vector Backbone:pFLAG-CMV3; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:15020428
Comments: The EGFR internal signal sequence was removed during the cloning of this construct. When compared to GenBank reference sequence NP_997538.1, the signal sequence corresponds to amino acids 1-24
Proper citation: RRID:Addgene_18788 Copy
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:7755; Vector Backbone:pcDNA3.1(-)/Myc-His B; Vector Types:Mammalian Expression; Bacterial Resistance:Chloramphenicol and Ampicillin
Defining Citation: PMID:18064608
Comments: pcDNA3.1(-)/myc-His B Gateway-compatible Destination vector. Combine with an Entry clone containing your gene of interest with the Gateway LR reaction to generate an expression clone containing an C-terminal myc-His tag.
Proper citation: RRID:Addgene_18701 Copy
Vector Backbone Description: Backbone Size:8718; Vector Backbone:pJG4-5; Vector Types:Yeast Expression, two-hybrid prey vector; Bacterial Resistance:Chloramphenicol and Ampicillin
Defining Citation: PMID:18064608
Comments: pJG4-5 Gateway-compatible Destination vector. Combine with an Entry clone containing your gene of interest with the Gateway LR reaction to generate a prey vector expression clone for yeast two-hybrid.
Proper citation: RRID:Addgene_18704 Copy
Vector Backbone Description: Backbone Size:12414; Vector Backbone:pEG202; Vector Types:Yeast Expression, two-hybrid bait vector; Bacterial Resistance:Chloramphenicol and Ampicillin
Defining Citation: PMID:18064608
Comments: pEG202 Gateway-compatible Destination vector. Combine with an Entry clone containing your gene of interest with the Gateway LR reaction to generate a bait vector expression clone for yeast two-hybrid.
Proper citation: RRID:Addgene_18705 Copy
Genetic Insert: ERK Activity Reporter
Vector Backbone Description: Backbone Marker:Promega; Backbone Size:4000; Vector Backbone:pCI; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:19033456
Proper citation: RRID:Addgene_18680 Copy
Species: Homo sapiens
Genetic Insert: Bicistronic fluorescent reporter gene with cap-dependent EYFP and IRES-dependent ECFP translation under the control of CMV promo
Vector Backbone Description: Backbone Marker:Clontech; Backbone Size:5300; Vector Backbone:pIRES2-EGFP; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:17023487
Comments: EYFP tagged at N-terminus with 3 copies of Myc epitope tags and at C-terminus with a copy of FLAG and polyhistidine (His6) epitope tags. ECFP tagged at the C-terminus with one copy of hemagglutinin (HA) and polyhistidine (His6) epitope tags.
EGFP of pIRES2-EGFP (Clontech) replaced at an 803 bp BstXI-NotI, PCR amplified fragment with ECFP from pECFP-Mito (Clontech) tagged at the C-terminus with a single copy of hemagglutinin (HA) and polyhistidine (His6) epitope tags; IRES-dependent translation yields ECFP-HA-His6 fusion protein. 895 bp Sac I/Eco RI fragment containing amino terminal 3 Myc epitope tags fused to EYFP obtained from PCR amplification of pEYFP-N1 (Clontech), which is also tagged at C-terminus with a single copy of FLAG and polyhistidine (His6) epitope tag; cap-dependent translation yields 3Myc-EYFP-Flag-His6 fusion protein.
See article for detailed information.
Proper citation: RRID:Addgene_18673 Copy
Species: Homo sapiens
Genetic Insert: NEDD8
Vector Backbone Description: Backbone Size:5446; Vector Backbone:pcDNA3; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:9353319
Comments: Note that this is the active form of NEDD8. Pro-NEDD8 (which is inactive) has an additional 5 amino acids at the c-terminus.
Proper citation: RRID:Addgene_18711 Copy
Species: Homo sapiens
Genetic Insert: ubiquitin
Vector Backbone Description: Backbone Size:5446; Vector Backbone:pcDNA3; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:9353319
Comments: This plasmid grows slowly. We encourage requesting scientists to grow for a longer period of time than normal. Please note that this plasmid runs as a dimer (>10kb) or multimer.
Proper citation: RRID:Addgene_18712 Copy
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