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  1. Choosing Your Perfect Empty Backbone

    Type
    Blog Post
    ...most popular mammalian expression vectors, c-Flag pcDNA3, comes from the pcDNA series. It has the CMV promoter...protein will allow you to visualize YGOI in vivo (try pcDNA3-EGFP). If you can get away with generating transiently...vectors (e.g. pBV-Luc) and gene silencing (e.g. pLKO.1 - TRC cloning vector). Check out our complete guide...

  2. Illuminating Epigenetics with A FRET Based Biosensor

    Type
    Blog Post
    ...reporter to target the K9 (pcDNA3-K9 histone methylation reporter) and K27 (pcDNA3-K27 histone methylation...treatment and improving patients lives. References 1. Ong-Abdullah, Meilina, et al. "Loss of Karma transposon...

  4. Arf GTPase Family

    Type
    Collection
    ...pET), Mammalian (pcDNA3), Gateway GTPase Arf3 377 181 Bacterial (pET), Mammalian (pcDNA3), Gateway GTPase...pET), Mammalian (pcDNA3), Gateway GTPase Arf5 381 180 Bacterial (pET), Mammalian (pcDNA3), Gateway GTPase...pET), Mammalian (pcDNA3), Gateway GTPase Arl3 403 182 Bacterial (pET), Mammalian (pcDNA3), Gateway GTPase... (pET), Mammalian (pcDNA3) GAP Arfgap2 84364 521 Bacterial (pET), Mammalian (pcDNA3) GAP Arfgap3 26286...Mammalian (pcDNA3, pEGFP) GAP Git2 9815 759 GAP ELMOD1 55531 334 Bacterial (pGEX4T), Mammalian (pcDNA3) GAP ...pGEX4T), Mammalian (pcDNA3) GAP ELMOD3 84173 381 Bacterial (pGEX4T), Mammalian (pcDNA3) GEF Arfgef1 10565...GTPase Arf6 382 175 Bacterial (pET), Mammalian (pcDNA3), Gateway GTPase Arl1 400 181 Bacterial (pET), Mammalian...

  5. Fluorescent Protein Guide: Empty Backbones

    Type
    Collection
    ...Bacterial Expression mRFP1 584 607 13 4.5 1 hr Monomer pcDNA3-mRFP - Mammalian Expression pMXs-mRFP1 - ...Expression ECFP 433 475 13 4.7 Prone to dimerization pcDNA3-CFP - Mammalian Expression pCAG-CFP - Mammalian...Expression EGFP 488 507 34 6 Prone to dimerization pcDNA3-EGFP - Mammalian Expression pCAG-GFP - Mammalian...Clover 505 515 84 6.1 30 min Prone to dimerization pcDNA3-Clover - Mammalian Expression pFA6a-link-yoClover-Kan...Plasmids EYFP 513 527 51 6.9 Prone to dimerization pcDNA3-YFP - Mammalian Expression Topaz 514 527 57 Prone...Expression mCitrine 516 529 59 5.7 Monomer (A206K) pcDNA3 mCitrine LIC cloning vector (6C) - Mammalian Expression...mRuby2 559 600 43 5.3 150 min Prone to dimerization pcDNA3-mRuby2 - Mammalian Expression pFA6a-link-yomRuby2...

  6. Empty Backbones - Choosing Your Perfect Plasmid Backbone

    Type
    Collection
    ...Localization pcDNA3-EGFP - C-terminal GFP for mammalian expression pcDNA3 GFP LIC cloning...AdEasier®-1 cells (strain) - Bacterial strain that contains AdEasy®-1 plasmid...Representative Empty Backbones Flag Epitope tag pcDNA3 Flag HA - N-terminal Flag-HA for... expression Puromycin Mammalian pLKO.1 - TRC cloning vector - Lentiviral... Tet-inducible lentiviral shRNA expression pMKO.1 puro - Retroviral shRNA expression ... expression Hygromycin Mammalian pLKO.1 hygro - Lentiviral shRNA expression... - Retroviral gene expression pMKO.1 GFP - Retroviral shRNA expression pSico...

  7. Neurodegeneration Plasmid Collection

    Type
    Collection
    ...Fawzi 127194 RP1B FUS 1-163 QQ4xSS #1 FUS His T7 ALS Nicolas Fawzi 127195 RP1B FUS 1-163 QQ4xSS #2 FUS His... type 1 scFv [L24/1] ITPR1 CMV Spinocerebellar ataxia James Trimmer 206790 IP3 receptor, type 1 scFv [...Lindquist 14121 pcDNA3-HtrA2-EGFP HTRA2 GFP CMV Parkinson's L. Miguel Martins 14122 pcDNA3-HtrA2-EGFP S306A...Mombaerts 15938 pcDNA3-HtrA2-FLAG HTRA2 Flag CMV Parkinson's L. Miguel Martins 15939 pcDNA3-HtrA2-FLAG S142D...Arrowsmith 65106 pcDNA3-ADAM10-HA ADAM10 HA CMV Alzheimer's Axel Ullrich 65107 pcDNA3-Delta(Pro-MP)ADAM10...Parkinson's Dori Woods 197146 pcDNA3-ApoE2 APOE CMV Alzheimer's Bradley Hyman 197147 pcDNA3-ApoE3 APOE CMV Alzheimer's...E320A) pcDNA3 CACNA1A CMV Spinocerebellar ataxia 6 Annette Dolphin 206125 Cav2.1 EIVA (E1707A) pcDNA3 CACNA1A...

  8. Luciferase Plasmid Collection

    Type
    Collection
    ...luciferase Eric Campeau 18964 Luciferase-pcDNA3 Firefly CMV Mammalian expression of firefly luciferase William...renilla luciferase Liqun Luo 51970 Nano-lantern/pcDNA3 Nano-lantern CMV Mammalian expression of Nano-lantern... Eric Campeau 21474 pLenti CMV V5-LUC Blast (w567-1) Firefly CMV Lentiviral expression of firefly luciferase... processes, biosensors, or optogenetic tools . pcDNA3.1+-pHlash , pRSETb-pHlash : A pH sensor created ...Gateway cloning. Lentiviral. Jorge Ferrer 87067 pcDNA3.1-ccdB-Nanoluc NanoLuc® Creation of N-terminal Nanoluc...Renilla luciferase is co-expressed. Oskar Laur 87070 pcDNA3.1-Nanoluc-ccdB NanoLuc® Creation of N-terminal Nanoluc...fusions using Gateway cloning Mikko Taipale 87065 pcDNA3.1-ccdB-Renilla Renilla Creation of C-terminal Renilla...

  9. 27 Hot Plasmids from 2016

    Type
    Blog Post
    ...a rate of up to 1 per 500-1000 bp. In comparison, the DNA replication error rate is 1 per 109 bp. Unlike...Transcriptional Output-TANGO (PRESTO-TANGO) to screen the NCC-1 library of approved drugs against the entire kit; ...generated 5 common 30bp overlap sequences (Sites #1-5). Each overlap site encodes a unique restriction...First, GMAP-compatible backbones for lentivirus LV 1-5 and retrovirus RV 2-5 were constructed. The authors...is multiplex automated genome engineering (MAGE) (1). MAGE uses recombineering (2) to simultaneously incorporate...of GFP (SEP) by a 9 amino-acid linker (see Figure 1). The key to the biosensor lies in pH: DsRed is relatively...inactivate SEP and leave only DsRed to fluoresce (1). With variants that can be targeted to specific cellular...

  10. Fluorescent Protein Guide: Subcellular Localization

    Type
    Collection
    ...Brown 11908 pEGFP-N1 alpha-actinin 1 Actin Filaments alpha-actinin 1 EGFP Carol Otey 27676 Cortactin-pmCherryC1...Huttenlocher 11908 pEGFP-N1 alpha-actinin 1 Focal Adhesions alpha-actinin 1 EGFP Carol Otey 31923 pPaxillin-PSmOrange...Autophagosome Sequestosome-1 EGFP Noboru Mizushima 38272 pMXs-IP GFP-WIPI-1 Autophagosome WIPI1 EGFP Noboru...apparatus eNOS(1-33) CFP Alexandra Newton 36205 pmTurquoise2-Golgi Golgi apparatus B4GALT1(1-61) mTurquoise2...EGFP Ikuo Wada 73208 pcDNA3.1-COXIV-COX8-dL5-2XG4S-mCer3 Mitochondria COX IV-derived (1-22 aa) import sequence... Shirihai 158003 pCMV-mGold-CAF1-C-10 Nucleus CAF-1 mGold Francois St-Pierre 27705 CAV1-mCherry Caveolae...Verkhusha 36208 pmTurquoise2-Mito Mitochondria COX8A(1-29) mTurquoise2 Dorus Gadella 26673 pEGFP-C1-Fibrillarin...

  11. CRISPR 101: Epigenetics and Editing the Epigenome

    Type
    Blog Post
    ...P2A-PuroR for lentiviral expression. Figure 1: dCas9-p300 adds H3K27ac marks to histones in promoter...lentiviral transduction. Lysine-specific Demethylase 1 (LSD1) Tatjana Sauka-Spengler's lab has deposited ...constructs including pcDNA-dCas9-p300 Core and pcDNA3.3-Nm-dCas9-p300 Core, as well as pLV-dCas9-p300-... in mammalian cells. This plasmid is used with pcDNA3.1-MS2-Tet1-CD to decrease methylation and activate...Methyltransferase MQ1 Margaret Goodell's lab has deposited pcDNA3.1-dCas9-MQ1(Q147L)-EGFP, a fusion of dCas9 to a ...

  12. Cre-lox system

    Type
    Collection
    ...Cre-GFP fusion EF-1 alpha Mammalian Sauer 11923 pBS598 EF1alpha-EGFPcre Cre-EGFP fusion EF-1 alpha Mammalian...mCherry coexpression EF-1 alpha AAV Deisseroth 55635 pAAV-EF1a-sCre SCre EF-1 alpha AAV Deisseroth 55636...55636 pAAV-EF1a-Cre Cre EF-1 alpha AAV Deisseroth 55638 pAAV-EF1a-vCre VCre EF-1 alpha AAV Deisseroth 59701...) EF-1 alpha AAV Cepko 69571 pAAV-EF1a-C-CreintG Cre recombinase dependent on GFP (CRE-DOG) EF-1 alpha...pAAV-EF1a-fDIO-Cre Cre EF-1 alpha AAV Engel 122518 pAAV-Syn-iCre iCre AAV Ritola 122960 pcDNA3.1_PA-Cre photo-activatable... Mammalian Sauer 11918 pBS513 EF1alpha-cre Cre EF-1 alpha Mammalian Sauer 11919 pBS448 RSV-GFPcre Cre-...11955 pBS505 EF1alpha-EGFPcre* Cre-EGFP fusion EF-1 alpha Mammalian Sauer 11956 pBS594 promoterless EGFPcre...

  13. Fujii Lab CRISPR Plasmids

    Type
    Collection
    ...gRNA-hIRF-1 #12 Expresses a guide RNA (gRNA) to target human IRF-1 promoter 61080 gRNA-hIRF-1 #12/pSIR ..._dCas9-2xAM_hIRF-1 Lentiviral plasmid expressing dCas9-2xAM and gRNA for human IRF-1 promoter. 98041 Sa-dCas9...interest. 98134 hIRF-1 gRNA #409 (Sa) Expresses a guide RNA (gRNA) to target human IRF-1 promoter for the ...system. 105283 hIRF-1 gRNA #351 (Sa) Expresses a guide RNA (gRNA) to target human IRF-1 promoter for the ...LexA are subjected to affinity purification (Fig. 1). In enChIP, specific genomic regions are tagged with...plasmids or browse the Fujii Lab web page . Figure 1: iChIP general scheme Figure 2: enChIP general scheme... Expresses a guide RNA (gRNA) to target human IRF-1 promoter 62190 3xFLAG-dCas9/pTEF1p-CYC1t Expresses...

  14. Genetic Code Expansion

    Type
    Collection
    ...Bacterial TAG Farren Isaacs 73545 pEvol-pAcFRS.1.t1 pAcFRS.1.t1 E. coli p-acetyl-l-phenylalanine (pAcF) Bacterial...Bacterial TAG Farren Isaacs 73547 pEvol-pAzFRS.1.t1 pAzFRS.1.t1 E. coli p-azido-l-phenylalanine (pAzF) Bacterial...7FTrp) Bacterial TAG Thomas Huber 207639 pEVOL-NBK-1 PylRS M. mazei Pyrrolysine Bacterial Cole DeForest...and tyrU gene replaced by GentR) and release factor 1. Expresses archaeal MjTyrRS/tRNA pair instead. For...L-lysine (TFA-Lys) Bacterial TAG Thomas Huber 182287 pcDNA3.1(+)_U6 tRNAPyl_CMV NESPylRS(AF) Y306A/Y384F (AF...PylRS M. mazei Bacterial TAA Wenshe Liu 182653 pcDNA3.1(+)_U6 tRNAPyl_CMV NESPylRS(AF)_IRES_eRF1(E55D)...

  15. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    ...light-sensitive LOV2 domain of Avena Sativa phototropin 1 (AsLOV2). In the dark, this peptide is caged by the...iLID nano and 800nM to 47µM for iLID micro; Figure 1). With this great affinity range, these LIDs have ...Figure 2). They are all available now at Addgene. 1 Lungu et al., Chem Biol. 2012 Apr 20;19(4):507-17....Guntas et al., Proc Natl Acad Sci USA 2015 Jan 6; 112(1):112-7. doi: 10.1073/pnas. CRISPR-Cas9 optogenetic... al., BMC Molecular Biology. 2013 August 20th; 14(1). TRICK: A method for visualizing the first round...for a variety of experimental purposes including (1) single or multiple, constitutive or inducible, gene...the years. The authors optimized these systems by: 1) decreasing the number of plasmids required for each...
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