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| Plasmid Name | Proper Citation | Insert Name | Organism | Bacterial Resistance | Defining Citation |
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pCR3-FLAG-Ubc13-C87A Resource Report Resource Website |
RRID:Addgene_43939 | ubiquitin-conjugating enzyme E2N | Homo sapiens | Ampicillin | PMID:17135271 | Backbone Size:5000; Vector Backbone:pCR3; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin | C87A | 2026-08-15 01:15:11 | 0 | |
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pCS2-USP44-HIS Resource Report Resource Website |
RRID:Addgene_43894 | USP44 | Homo sapiens | Ampicillin | PMID:17443180 | Backbone Size:4700; Vector Backbone:p142; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin | 2026-08-15 01:15:11 | 0 | ||
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pFS217-USP44_MSCV-TAP Resource Report Resource Website |
RRID:Addgene_43896 | USP44 | Homo sapiens | Ampicillin | PMID:17443180 | Backbone Size:7556; Vector Backbone:MSCV; Vector Types:Mammalian Expression, Retroviral; Bacterial Resistance:Ampicillin | 2026-08-15 01:15:10 | 0 | ||
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pGEX4T1-TRAF6-C70A Resource Report Resource Website |
RRID:Addgene_43930 | TNF Receptor-Associated Factor 6 | Ampicillin | PMID:17135271 | Backbone Marker:GE Healthcare; Backbone Size:4910; Vector Backbone:pGEX4T1; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin | C70A | 2026-08-15 01:15:11 | 0 | ||
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pTF258 (pFA6a-PSc-6xGly-His3MX) Resource Report Resource Website |
RRID:Addgene_44062 | Ampicillin | The depositing lab has constructed and deposited here 36 plasmids that can be used to generate linear PCR products to fuse to the 3' end of an ORF in the yeast genome and add (TEV or PreScission protease site) - 6X Gly linker - (12X His, 2X Strep, 3X FLAG, Protein A, or V5) - ADH1 transcription terminator - (KanMX, HphMX, or His3MX). Six plasmids (Addgene plasmids 44062, 44068, 44074, 44080, 44086, and 44092) have no epitope/purification tag and so can be used to add an alternate tag by standard cloning into unique XhoI-KpnI-XbaI sites, if desired. The other 30 plasmids are all of the possible combinations of the protease site - tag - marker options listed above. After fusion, the modified protein can then be followed with the epitope or purified using the tag and released from the matrix with the protease site. The constructs were designed with one of two different protease sites to allow a complex with two different tags to be purified in two successive steps, using each protease to release the complex from each purification matrix. The schematic diagram (see attached above) represents the format of the constructs and shows the proper scheme for producing the desired PCR products (see also below). If only epitope tagging is needed, the upstream primer can be adjusted to skip the protease site (see example below). Construction: DS5421 (pFA6a-6xGLY-3XFLAG-HIS3MX6 (Addgene Plasmid #20753), DS5422 (pFA6a-6xGLY-3XFLAG-KanMX) (Addgene Plasmid #20754), or DS5423 (pFA6a-6xGLY-3XFLAG-HphMX) (Addgene Plasmid #20755) (corresponding to each selection marker) was digested with AscI and PacI to release 3XFlag and replace it with TEV or PreScision sites followed by 6xGly and an XhoI-KpnI-XbaI linker. Affinity tags were constructed by PCR or by annealing oligos to fit into the XhoI-XbaI gap. This includes 12xHis, 2xStrep II, 3xFLAG, PrtA (2 repeats as found in pBS1479), and V5. Each tag was inserted into each base vector, making 30 templates with all combinations of protease site, tag, and selectable marker. PCR amplification: These two primers (along with 50 nt of homology to the targets at the 5' ends) will amplify the desired product from any of the constructs (the reading frame for P1 is important – alternating codons bracketed). P1 5’-(homology)...[CGA]CGG[ATC]CCC[GGG]TTA[ATT]AAC-3’ P2 5’-(homology)...GATATCATCGATGAATTCGAGCTCGTTT-3’ If only epitope tagging is needed, the upstream primer can be adjusted to skip the protease site. As an example, for the V5-tag-containing constructs, usage of the following primer (5’-(homology)... GGTGGAGGCTCTAGAGGTAAGCCAATTCCA-3’) in place of P1 will omit the protease cleavage site and add only 3xGly and V5 (GGGSR-V5-Stop). | Backbone Marker:Mark Hochstrasser lab (Yale Univeristy, New Haven, CT) (PMID: 19243080); Backbone Size:4142; Vector Backbone:pFA6a-6xGLY-3xFLAG-HIS3MX6 (Addgene plasmid #20753); Vector Types:PCR-based yeast C-terminal tagging; Bacterial Resistance:Ampicillin | 2026-08-15 01:15:12 | 0 | ||||
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pCAG-LplA(AILR,36)-NRX3b D137A Resource Report Resource Website 1+ mentions |
RRID:Addgene_43920 | AILR-LplA36-NRX3b | Homo sapiens | Ampicillin | PMID:23457442 | Backbone Size:4247; Vector Backbone:pCAG; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin | D137A | 2026-08-15 01:15:11 | 1 | |
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pXL-CAG-mBirA272-NRX3b Resource Report Resource Website |
RRID:Addgene_43921 | BirA272-NRX3b | Homo sapiens | Ampicillin | PMID:23457442 | 272 indicates the amino acid position that the BirA is inserted in for NRX3b. There is a single alanine deletion in BirA, which is a non-dimerizing mutation. The BirA and AILR insert regions of the plasmid is codon optimized. | Backbone Size:6299; Vector Backbone:pXL-CAG; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin | See comments | 2026-08-15 01:15:10 | 0 |
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pXL-CAG-mBirA272-NRX3b D137A Resource Report Resource Website |
RRID:Addgene_43922 | BirA272-NRX3b | Homo sapiens | Ampicillin | PMID:23457442 | Backbone Size:6299; Vector Backbone:pXL-CAG; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin | D137A | 2026-08-15 01:15:11 | 0 | |
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pXL-CAG-3XAP-NLG1 Resource Report Resource Website |
RRID:Addgene_43923 | 3XAP-NLG1 | Rattus norvegicus | Ampicillin | PMID:23457442 | Backbone Size:6390; Vector Backbone:pCAG; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin | 2026-08-15 01:15:11 | 0 | ||
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pCR3-FLAG-TRAF6-C70A Resource Report Resource Website 1+ mentions |
RRID:Addgene_43926 | TNF Receptor-Associated Factor 6 | Mus musculus | Ampicillin | PMID:17135271 | Backbone Marker:Invitrogen; Backbone Size:5000; Vector Backbone:pCR3; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin | C70A | 2026-08-15 01:15:11 | 1 | |
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MAP2-GCaMP3 Resource Report Resource Website 1+ mentions |
RRID:Addgene_43917 | GCaMP3 | Synthetic | Ampicillin | PMID:22174877 | The MAP2-GFP lentiviral reporter plasmid pGZ-hMAP2 was purchased from System Biosciences (SR10047PA-1). The MAP2 promoter sequence from this plasmid was then cloned along with the Gateway cassette of pEF-DEST51 into the backbone of FUdeltaGW-rtTA (Addgene plasmid 19780) following removal of the ubiquitin promoter-rtTA sequence to produce FU-MAP2-Gateway (Addgene plasmid #43915). This destination vector was subsequently used to generate MAP2-GCaMP3 (GCaMP3 cloned from G-CaMP3, Addgene plasmid 22692). The V5 and His tags present in the vector backbone are not expressed with the GCaMP3 insert. | Backbone Marker:Addgene plasmid 43915; Backbone Size:9600; Vector Backbone:FU-MAP2-Gateway; Vector Types:Mammalian Expression, Lentiviral; Bacterial Resistance:Ampicillin | E247V in GCaMP3 | 2026-08-15 01:15:11 | 1 |
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pDL96 Resource Report Resource Website |
RRID:Addgene_43910 | DIF1 | Saccharomyces cerevisiae | Ampicillin | PMID:18851834 | Backbone Marker:Stratagene; Backbone Size:4898; Vector Backbone:pRS416; Vector Types:Yeast Expression; Bacterial Resistance:Ampicillin | AAEAAA | 2026-08-15 01:15:11 | 0 | |
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pDL127 Resource Report Resource Website |
RRID:Addgene_43911 | DIF1 | Saccharomyces cerevisiae | Ampicillin | PMID:18851834 | Backbone Marker:Stratagene; Backbone Size:4898; Vector Backbone:pRS416; Vector Types:Yeast Expression; Bacterial Resistance:Ampicillin | HUG domain mutated VGMRIRQ->AAAAAAA, S59, Q66H | 2026-08-15 01:15:10 | 0 | |
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pDL128 Resource Report Resource Website |
RRID:Addgene_43912 | DIF1 | Saccharomyces cerevisiae | Ampicillin | PMID:18851834 | Backbone Marker:Stratagene; Backbone Size:4898; Vector Backbone:pRS416; Vector Types:Yeast Expression; Bacterial Resistance:Ampicillin | SML domain mutated PPMLINQRT->AAAAAAAA, S59G, Q66H | 2026-08-15 01:15:10 | 0 | |
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FU-MAP2-Gateway Resource Report Resource Website |
RRID:Addgene_43915 | Chloramphenicol and Ampicillin | PMID:22174877 | The MAP2-GFP lentiviral reporter plasmid pGZ-hMAP2 was purchased from System Biosciences (SR10047PA-1). The MAP2 promoter sequence from this plasmid was then cloned along with the Gateway cassette of pEF-DEST51 into the backbone of FUdeltaGW-rtTA (Addgene plasmid 19780) following removal of the ubiquitin promoter-rtTA sequence to produce FU-MAP2-Gateway. | Backbone Marker:Addgene plasmid 19780; Backbone Size:9600; Vector Backbone:FUdeltaGW-rtTA; Vector Types:Mammalian Expression, Lentiviral, Gateway Cloning; Bacterial Resistance:Chloramphenicol and Ampicillin | 2026-08-15 01:15:10 | 0 | |||
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pCoofy6 Resource Report Resource Website 1+ mentions |
RRID:Addgene_43990 | Kanamycin | PMID:23410102 | Test each preparation of the plasmid by transforming it into non-resistant cells (ex. DH5a) to ensure negative selection by ccdB kills all the transformed cells as expected. | Backbone Marker:Novagen, EMBL; Backbone Size:6657; Vector Backbone:pET, pET28M-Sumo3; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin | 2026-08-15 01:15:11 | 1 | |||
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pCoofy17 Resource Report Resource Website |
RRID:Addgene_43991 | Kanamycin | PMID:23410102 | Test each preparation of the plasmid by transforming it into non-resistant cells (ex. DH5a) to ensure negative selection by ccdB kills all the transformed cells as expected. | Backbone Marker:Novagen; Backbone Size:6669; Vector Backbone:pET, pCoofy6; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin | 2026-08-15 01:15:12 | 0 | |||
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pCoofy22 Resource Report Resource Website |
RRID:Addgene_43992 | Kanamycin | PMID:23410102 | Test each preparation of the plasmid by transforming it into non-resistant cells (ex. DH5a) to ensure negative selection by ccdB kills all the transformed cells as expected. | Backbone Marker:Novagen; Backbone Size:5764; Vector Backbone:pET, pCoofy7; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin | 2026-08-15 01:15:11 | 0 | |||
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pCoofy31 Resource Report Resource Website |
RRID:Addgene_43993 | Kanamycin | PMID:23410102 | C-terminal Tag has 6 His residues, LP2 primer needs to be adapted to 6His Test each preparation of the plasmid by transforming it into non-resistant cells (ex. DH5a) to ensure negative selection by ccdB kills all the transformed cells as expected. | Backbone Marker:Novagen; Backbone Size:5808; Vector Backbone:pET, pCoofy12; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin | 2026-08-15 01:15:11 | 0 | |||
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pCoofy36 Resource Report Resource Website |
RRID:Addgene_43994 | Kanamycin | PMID:23410102 | Test each preparation of the plasmid by transforming it into non-resistant cells (ex. DH5a) to ensure negative selection by ccdB kills all the transformed cells as expected. | Backbone Marker:Novagen; Backbone Size:5845; Vector Backbone:pET, pCoofy34; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin | 2026-08-15 01:15:12 | 0 |
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