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http://www.addgene.org/32520

Species: Rattus norvegicus
Genetic Insert: Constitutively active Rheb (S16H)
Vector Backbone Description: Backbone Marker:Addgene plasmid #32519; Vector Backbone:pHAGE-CMV-Rheb-IRES-eGFP-W; Vector Types:Mammalian Expression, Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:20062052
Comments: The constitutively activating S16H mutation was introduced to pHAGE-CMV-Rheb-IRES-eGFP-W (Addgene plasmid #32519) by site directed mutagenesis. The original pHAGE-CMV-IRES lentiviral vector is from Dr Richard C. Mulligan at Harvard (Mostoslavsky G et al., 2005).

Proper citation: RRID:Addgene_32520 Copy   

•   Source: Addgene

http://www.addgene.org/32481

Species: Homo sapiens
Genetic Insert: PSAML141F,Y115F-GlyR
Vector Backbone Description: Backbone Size:5100; Vector Backbone:AAV2; Vector Types:AAV, Cre/Lox, Adeno-associated virus, FLEX switch; Bacterial Resistance:Ampicillin
Defining Citation: PMID:21885782
Comments: Comments: These rAAV vectors are prone to recombination. This is a well known issue with these rAAV vectors and is due to the inverted terminal repeats (ITRs) required for rAAV production. To minimize recombination, we propagate these plasmids in NEB Stable cells. Also, to minimize recombination, cells should be cultured at 30 C. Note that these cultures will grow slowly (20 h for minipreps). Better yields and culture times are obtained with 2xYT as the media. This is strongly recommended. Because recombination may still happen, we do a panel of restriction digestions to assess whether the ITRs are in tact. Separate digestions with Sma1 and Msc1 should be performed. The expected patterns can be calculated from the attached sequence. In Magnus et al, the PSAMY115F,L141F-GlyR channel was emphasized for silencing because this channel has the lowest acetylcholine responsiveness. However, PSAML141F-GlyR constructs, lacking the Y115F mutation, already have low ACh potency. The Y115F mutation also slightly increases the EC50 for the activator ligand, PSEM89S. Since PSAML141F-GlyR constructs already have very low acetylcholine potency, in many cases it may not be worth the slight reduction in PSEM potency. We have made both constructs available for researchers who may find that they have different requirements with respect to acetylcholine responsiveness.

Proper citation: RRID:Addgene_32481 Copy   

•   Source: Addgene

http://www.addgene.org/32475

Species: Homo sapiens
Genetic Insert: PSAMQ79G,Q139G-5HT3HC
Vector Backbone Description: Backbone Size:5100; Vector Backbone:AAV2; Vector Types:AAV, Cre/Lox, Adeno-associated virus, FLEX switch; Bacterial Resistance:Ampicillin
Defining Citation: PMID:21885782
Comments: These rAAV vectors are prone to recombination. This is a well known issue with these rAAV vectors and is due to the inverted terminal repeats (ITRs) required for rAAV production. To minimize recombination, we propagate these plasmids in NEB Stable cells. Also, to minimize recombination, cells should be cultured at 30 C. Note that these cultures will grow slowly (20 h for minipreps). Better yields and culture times are obtained with 2xYT as the media. This is strongly recommended. Because recombination may still happen, we do a panel of restriction digestions to assess whether the ITRs are in tact. Separate digestions with Sma1 and Msc1 should be performed. The expected patterns can be calculated from the attached sequence.

Proper citation: RRID:Addgene_32475 Copy   

•   Source: Addgene

http://www.addgene.org/32468

Genetic Insert: G-GECO1.2
Vector Backbone Description: Backbone Size:4200; Vector Backbone:Customized Vector; Vector Types:Bacterial Expression, Lab constructed; Bacterial Resistance:Ampicillin
Defining Citation: PMID:21903779
Comments: The vector is modified from pBAD/ His B (Invitrogen).

Proper citation: RRID:Addgene_32468 Copy   

•   Source: Addgene

http://www.addgene.org/32465

Species: synthetic construct
Genetic Insert: R-GECO1.0
Vector Backbone Description: Backbone Size:4200; Vector Backbone:Customized Vector; Vector Types:Bacterial Expression, Lab constructed; Bacterial Resistance:Ampicillin
Defining Citation: PMID:21903779
Comments: The vector is modified from pBAD/ His B (Invitrogen).

Proper citation: RRID:Addgene_32465 Copy   

•   Source: Addgene

http://www.addgene.org/32505

Species: Homo sapiens
Genetic Insert: CIB1
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:5400; Vector Backbone:pcDNA3.1; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:11756406

Proper citation: RRID:Addgene_32505 Copy   

•   Source: Addgene

http://www.addgene.org/32574

Species: Homo sapiens
Genetic Insert: p66shc
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:5500; Vector Backbone:pcDNA3.1 His; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:11884717
Comments: This plasmid is a corrected version of pcDNA3.1his p66shc (Addgene plasmid 10972 http://www.addgene.org/10972). D2N and S60P mutations present in the original sample have been corrected in this plasmid.

Proper citation: RRID:Addgene_32574 Copy   

•   Source: Addgene

http://www.addgene.org/32577

Species: Mus musculus
Genetic Insert: CAMKIIalpha promoter 1.3kb version
Vector Backbone Description: Backbone Marker:Invitrogen; Vector Backbone:pDONR221 P1-P4; Vector Types:Gateway Cloning; Bacterial Resistance:Kanamycin
Defining Citation: PMID:21772812

Proper citation: RRID:Addgene_32577 Copy   

•   Source: Addgene

http://www.addgene.org/32610

Genetic Insert: Tcf/Lef:H2B-GFP
Vector Backbone Description: Backbone Size:4700; Vector Backbone:pEGFP-N1; Vector Types:; Bacterial Resistance:Kanamycin
Defining Citation: PMID:21176145
Comments: To g ene r a t e t h e p T CF /L e f :H 2 B -GFP co n s t ru c t , s i x copies of the TCF/Lef response elements together with the hsp68 minimal promoter from the TCF/Lef-LacZ reporter construct [11] were inserted into the AseI/Nhe1 sites of pCMV::H2B-GFP.

Proper citation: RRID:Addgene_32610 Copy   

•   Source: Addgene

http://www.addgene.org/32698

Species: Homo sapiens
Genetic Insert: RAP1
Vector Backbone Description: Vector Backbone:pAXEF-1; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin and Tetracycline
Defining Citation: PMID:15469846
Comments: Q63 changed to E to give phenotype

Proper citation: RRID:Addgene_32698 Copy   

•   Source: Addgene

http://www.addgene.org/32614

Species: Gallus gallus
Genetic Insert: 1XDR4
Vector Backbone Description: Backbone Marker:Mark Emerson; Backbone Size:6610; Vector Backbone:Stagia3; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:21060789
Comments: Note that there are some minor discrepancies between Addgene's quality control sequence and the depositor's assembled sequence. These differences should not affect plasmid function.

Proper citation: RRID:Addgene_32614 Copy   

•   Source: Addgene

http://www.addgene.org/32612

Species: Gallus gallus
Genetic Insert: Thyroid Response Element
Vector Backbone Description: Backbone Marker:Mark Emerson; Backbone Size:6610; Vector Backbone:Stagia3; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:21060789

Proper citation: RRID:Addgene_32612 Copy   

•   Source: Addgene

http://www.addgene.org/32600

Genetic Insert: mRFP1
Vector Backbone Description: Backbone Size:4700; Vector Backbone:pCAGGS; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:15996270
Comments: The mRFP1 coding sequence was amplified by PCR using primers 5'RFP (5'-cgtagaattcgccaccaatggctagcatgactgg) and 3'RFP (5'-gcacgaattcgggcgccggtggagtggcggcc) using Pfx Polymerase (Invitrogen). The resulting product was cloned into the EcoRI site of pCAGGS to generate pCXmRFP1. Transient transfection of Cos-7 cells using Fugene 6 Transfection Reagent as per manufacturer's recommendations (Roche) was used to evaluate pCX-mRFP1, and verify that it produced robust red fluorescence More Information: https://www.ncbi.nlm.nih.gov/pubmed/12060735 A monomeric red fluorescent protein. Campbell et al (PNAS. 2002 Jun 11; 99:12)

Proper citation: RRID:Addgene_32600 Copy   

•   Source: Addgene

http://www.addgene.org/32601

Genetic Insert: GPI-GFP
Vector Backbone Description: Backbone Size:4700; Vector Backbone:pCAGGS; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:16604528

Proper citation: RRID:Addgene_32601 Copy   

•   Source: Addgene

http://www.addgene.org/32602

Genetic Insert: myr-Venus
Vector Backbone Description: Backbone Size:4700; Vector Backbone:pCAGGS; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:16604528
Comments: To generate myristoylated fluorescent fusion proteins, an N-terminal myristoylation tag derived from Src was generated by adding the sequences MGSSKSKPK to the N-terminus of any given fluorescent protein. This was achieved by using the following oligonucleotide: 5′ GFP-MYR-Eco (5′-CTT GAA TTC GCC ACC ATG GGA AGC AGC AAG AGC AAG CCA AAG GTG AGC AAG GGC GAG GAG CTG). The GFP and Venus coding sequences were amplified from pEGFP-N1 (BD Biosciences, San Jose, CA) and pCS2-Venus (Nagai et al., 2002) to generate myr-GFP and myr-Venus, respectively, by high-fidelity polymerase chain reaction (PCR) using Pfx Polymerase (Invitrogen, La Jolla, CA) with the 5′ myristoylation primer combined with a 3′-GFP primer (5′-GTC ATG AAT TCT TAC TTG TAC AGC TCG TCC) primer, respectively. The resulting product was cloned into the EcoRI site of pCAGGS to generate pCX::myr-EGFP and pCX::myr-Venus

Proper citation: RRID:Addgene_32602 Copy   

•   Source: Addgene

http://www.addgene.org/32669

Species: Mus musculus
Genetic Insert: Kir2.1-mCherry
Vector Backbone Description: Backbone Marker:Invitrogen; Vector Backbone:pDONR221 P5-P2; Vector Types:Gateway Cloning; Bacterial Resistance:Kanamycin
Defining Citation: PMID:21772812

Proper citation: RRID:Addgene_32669 Copy   

•   Source: Addgene

http://www.addgene.org/32650

Genetic Insert: FLPo
Vector Backbone Description: Backbone Marker:Addgene plasmid 32634; Backbone Size:15428; Vector Backbone:pSADdeltaG; Vector Types:Mammalian Expression, Rabies virus; Bacterial Resistance:Ampicillin
Defining Citation: PMID:21867879

Proper citation: RRID:Addgene_32650 Copy   

•   Source: Addgene

http://www.addgene.org/32641

Species: Mus musculus
Genetic Insert: Kir2.1
Vector Backbone Description: Backbone Marker:Promega; Vector Backbone:pGEM-T Easy; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:15157418
Comments: Addgene's sequencing results identified a single nucleotide insertion at bp#60/61 when compared to the author's sequence. The insert is upstream of the coding region and should not be a concern for the intended purpose of the plasmid. Silent mutations in Kir2.1 that do not affect the amino acid sequence were also found in Addgene's sequencing results.

Proper citation: RRID:Addgene_32641 Copy   

•   Source: Addgene

http://www.addgene.org/32642

Species: Mus musculus
Genetic Insert: tetO-Kir2.1-IRES-TLZ
Vector Backbone Description: Backbone Marker:Stratagene; Vector Backbone:pBlueScript; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:15157418
Comments: Addgene's sequencing results identified a single nucleotide a single nucleotide deletion at bp#1135 when compared to the author's sequence. The insert is upstream of the coding region and should not be a concern for the intended purpose of the plasmid. Silent mutations in Kir2.1 that do not affect the amino acid sequence were also found in Addgene's sequencing results.

Proper citation: RRID:Addgene_32642 Copy   

•   Source: Addgene

http://www.addgene.org/32646

Genetic Insert: ChR2
Vector Backbone Description: Backbone Marker:Addgene plasmid 32634; Backbone Size:15428; Vector Backbone:pSADdeltaG; Vector Types:Mammalian Expression, Rabies virus; Bacterial Resistance:Ampicillin
Defining Citation: PMID:21867879

Proper citation: RRID:Addgene_32646 Copy   

•   Source: Addgene