...tag with TEV cleavage, AmpR 26093 pET15-MHL EF456738 Hexahistidine tag with TEV cleavage, AmpR 26094 pET28a-LIC...with Thrombin cleavage, KanR 26096 pET28-MHL EF456735 Hexahistidine tag with TEV cleavage, KanR 26098 pCW-LIC...with TEV cleavage, KanR 26105 pNIC-CTHF EF199844 Hexahistidine tag, FLAG tag with TEV cleavage, KanR 26106...thioredoxin with TEV cleavage, KanR 26107 pNIC-ZB GU452710 Hexahistidine tag , Z-basic, TEV cleavage, AmpR 26108...pET28a-LIC EF442785 Hexahistidine tag with Thrombin cleavage, KanR 26101 pET28GST-LIC EF456739 GST-tag and ...pFBOH-LIC EF456740 Hexahistidine tag with Thrombin cleavage, Bac-to-Bac Baculovirus Expresion vector 26117...pFB-LIC-Bse EF199842 Hexahistidine tag with TEV cleavage, AmpR 26095 pFHMSP-LIC-N N/A Expression vector...

.../IPTG inducible) N terminal, TEV cleavable 6x-His tag Ophear Gileadi pET-28-a based expression vector ...Luciano Marraffini Bacterial expression of Cas9 nuclease gRNA. For a more comprehensive list of our bacterial...TALEN fused to an E. coli codon optimized FokI endonuclease domain. pET-Sangamo-His 40787 TALEN Gang Bao...TALEN fused to an E. coli codon optimized FokI endonuclease domain. Return to Top Visualization and Tagging...in bacteria, add epitope tags to it, and, later, cleave those tags from the purified protein. Protein purification...challenging, but once you get it right, you can learn many things about your protein’s function, structure...lactose/IPTG inducible) N terminal GST tag with TEV cleavage site to remove tag Nicola Burgess Brown pGEX expression...

...glycosylase, aka T4 Endonuclease V) 165540 pOpen-T7EI T7 Endonuclease I 165551 pOpen-Endonuclease IV E. coli E... E. coli Endonuclease IV 165561 pOpen-Endonuclease VIII E. coli E. coli Endonuclease VIII 165571 pOpen-Exonuclease...pOpen-Exonuclease III E. coli E. coli Exonuclease III 165559 pOpen-T5gene12/13 T5 Exonuclease 165560 pOpen-T4...Collection DNA Ligases DNA Polymerases Restriction Endonucleases Reverse Transcriptases RNA Polymerases Other...below: DNA Ligases DNA Polymerases Restriction Endonucleases Reverse Transcriptases RNA Polymerases Other...phi29 DNA Polymerase Return to top Restriction Endonucleases Addgene ID Plasmid Gene/insert 165504 pOpen-...

...mCherry-LaminA-C-18 Nuclear Envelope LaminA1 mCherry* Michael Davidson 98831 mScarlet-I-LaminB Nuclear Envelope ...mTurquoise2-LaminB Nuclear Envelope LaminB mTurquoise2 Dorus Gadella 55069 mCherry-LaminB1-10 Nuclear Envelope ...nucleoplasm None dL5 FAP; mCerulean3 Marcel Bruchez 36206 pmTurquoise2-NES Non-nucleus Nuclear Export Sequence ...Non-nucleus Nuclear Export Sequence mScarlet-I Dorus Gadella 85061 pmScarlet-H_NES_C1 Non-nucleus Nuclear Export...sequence and COX VIII signal peptide dL5 FAP; mCerulean3 Marcel Bruchez 44385 pLV-mitoGFP Mitochondria...2xG4S-mCer3 Nucleus NLS (from Mak16p protein) dL5 FAP; mCerulean3 Marcel Bruchez 73207 pcDNA3.1-KozATG-dL5-2XG4S-mCer3...Dorus Gadella 85060 pmScarlet_NES_C1 Non-nucleus Nuclear Export Sequence mScarlet Dorus Gadella 37351 pQC...

...DS-SPcasN- Bacterial nuclease-null SP Cas9 expression 48659 DS-ST1casN- Bacterial nuclease-null ST1 Cas9 expression...expression of a Cas9 protein bearing a C terminus SV40 nuclear localization signal with one or more guide RNAs... III promoter. Cas9 unwinds the DNA duplex and cleaves both strands upon recognition of a target sequence...above Cas9 protein, NM is comparably active as a nuclease and a transcriptional activator. Streptococcus...slightly higher activities than the others as a nuclease and as an activator, but is more restricted in...transcriptional repressor. All genes except the SP nuclease are human codon optimized but function well in...SP and ST1. Protospacer-adjacent motifs (PAMs): Please note that NM, ST1, and TD recognize different PAMs...

...targets may not result in equivalent cleavage efficiency. In fact, cleavage efficiency may increase or decrease...are denatured-renatured, treated with a nuclease that cleaves DNA heteroduplexes, and run on an agarose...decrease expression of a target gene? Once you have a clear understanding of your experimental goal, you are...Infects dividing and non-dividing cells AAV is the least toxic method for in vivo viral delivery RNA delivery... sequence and target DNA may result in reduced cleavage. The number of alleles for each gene may vary ...likely to result in the most efficient on-target cleavage. The gRNA target sequence needs to match the target...gRNA design. In general, off-target sites are not cleaved as efficiently when mismatches occur near the PAM...

...Expression Cerulean 433 475 27 4.7 Monomer (A206K) Cerulean - Mammalian Expression pCS2+8CCerulean - Zebrafish...Expression mCerulean C1 - Mammalian Expression mCerulean3 433 475 32 3.2 Monomer (A206K) mCerulean3-N1 - Mammalian...Expression mCerulean3-C1 - Mammalian Expression moxCerulean3 434 474 36 3.2 Monomer (A206K) moxCerulean3 - Mammalian...C.elegans/Sea urchin pCS2+8NCerulean - Zebrafish/Xenopus/C.elegans/Sea urchin mCerulean N1 - Mammalian Expression...Plasmids - Includes tagging with mCherry, mCitrine, mCerulean Davidson Lab Plasmids - Includes many N- and C-terminal... Plasmids - Set includes tagging with mCherry, Cerulean, Citrine, and eGFP pPD95_75 - C-terminal GFP for...cloning; includes tagging with ECFP, EYFP, DsRed, Cerulean Bacteria Gradia Lab Bacterial Vectors - Includes...

...Looking For? Please let us know what you would like to see available to industry scientists. Please note this...page for more information. To learn more about Addgene’s MTA process please visit our Help Center . The ...collection of plasmids that are available to industry. Learn more about the available plasmids and how to order...Addgene as well as by our trusted partner labs. Learn More ID Recombinant Antibody Reactivity Recommended...account, you can place orders through our website. Please note that all orders must be placed online through... product pages after you login to your account. Please see our Help Center for more information on pricing...language which we are allowed to use for distribution. Please see the Sample Agreements on our Technology Transfer...

...Looking For? Please let us know what you would like to see available in viral format. Please note this does...available in each of the above serotypes and buffers. Please click on the plasmid name for ordering information...the AAV2(Y444F) serotype in future publications, please acknowledge Arun Srivastava and cite Petrs-Silva...the AAV2(trpYF) serotype in future publications, please acknowledge Arun Srivastava and cite Petrs-Silva...AAV2(4pMut)dHS serotype in future publications, please acknowledge Shannon Boye and cite Boye, et al. ...AAV6(dbY-F+T-V) serotype in future publications, please acknowledge Arun Srivastava and cite Ling, et al...using the AAV44.9 serotype in future publications, please acknowledge John Chiorini and Shannon Boye and ...

...have final approval before anything is released publicly, but please only share materials that you are comfortable...Learn about Addgene's 2018 AAV Education and Development (AAVED) workshop series. The...Leila Haery from Addgene will also be speaking or leading workshops. Tentative Meeting Agenda Thursday, September...examples of AAV strategies for intersectional control. Please see our list of suggested materials below. Suggested...Detailed protocol: AAV Production in HEK293 Cells Please share materials by uploading to your Dropbox link...comfortable and willing to share. Please only share materials that you have permission to share (e.g.,...contributions and your commitment to open science! Please see the meeting preparation section for how to ...

...partnership and the plasmids that are available, please see the Ginkgo Bioworks Blog post: Synthesizing...information on ordering from a for-profit entity, please see our Plasmid Distribution to Industry page. ...entry into cells. ( 2 ) FURIN - an enzyme that cleaves precursor proteins to a biologically active state... The SARS-CoV-2 S protein contains a potential cleavage site for furin proteases.( 3 ) BSG - (CD147), ...function.( 5 ) TACE - (ADAM17) a metalloprotease that cleaves hACE2, potentially increasing viral infectivity...degradation to develop the method termed DNA Endonuclease Targeted CRISPR Trans Reporter (DETECTR). Mammoth...resources, such as reagents, data, news, and more, please see Ginkgo's Synthetic Biology (and beyond) Community...

...CaMKII-somBiPOLES-mCerulean CamKII Bidirectional Optogenetics Wiegert 154951 hSyn-DIO-somBiPOLES-mCerulean Syn Bidirectional...Looking For? Please let us know what you would like to see available in viral format. Please note this does...DREADDs Activators Inhibitors Other Molecular Tools Clear Filters ID Name Promoter Activity Category PI 37825...GCaMP6f-P2A-nls-dTomato Syn GCaMP6f and physically separate nuclear dTomato Calcium sensor Ting 51083 AAV-EF1a-DIO-...AAV-EF1a-DIO-GCaMP6f-P2A-nls-dTomato EF1a GCaMP6f and physically separate nuclear dTomato, Cre-dependent Calcium sensor Ting 83899...GCaMP6s-P2A-nls-dTomato Syn GCaMP6s and physically separate nuclear dTomato Calcium sensor Ting 100853 pAAV.Syn.Flex.NES-jRGECO1a.WPRE.SV40...available items found to match all requested criteria. Please consider using fewer filtering options. Browse ...

...a zinc finger (ZF) nuclease. Zinc fingers recognize triplets and the FokI nuclease operates as a dimer...sites. (B) A TAL effector nuclease (TALEN). Similar in principle to the ZF nuclease, the components of the...these tools and the researchers who are learning to wield them. Learn more about Addgene’s TAL effector and...years of research has shown that when fused to a nuclease or activation domain, a ZF array is an effective...at Massachusetts General Hospital also recently released a TALEN kit through Addgene. This kit, first described...

...amplify. Typically use primers at least 100 bp from the predicted cleavage site to ensure detection would...322.2 KB If you have trouble loading this video, please try viewing on YouTube . Abstract The prokaryotic...lines. Protocol For referenced Tables and Figures, please access the manuscript PDF posted at the top of ...genomic matches or near-matches to reduce risk of cleavage away from target sites (off-target effects). Ensure...which are likely to cause frameshift mutations leading to nonsense mediated decay of the mRNA transcript...of primers upstream and downstream of the sgRNA cleavage sites (“deletion band”; Figures 1 - 2 ). In the...for scarring (small indels produced at the sgRNA cleavage site without the intended deletion). Use a pair...

...code inserted into the plasmid. This produces the least amount of variation between antibodies, allows for... induced to express antibodies. Antibodies are released into the growth medium and can either be used ...fragments - Fab fragments are created by enzymatically cleaving the variable regions of an antibody (each Fab ...they should be stored at -20 °C in aliquots of at least 10 µL. Antibodies cannot tolerate freeze-thaw cycles...single cells. Includes: Flow cytometry Read on to learn more about the applications common to each category...should be run with your protein of interest and at least one ubiquitously expressed protein such as GAPDH...protein:DNA complexes. After capture, the DNA is released from the complex and can be sequenced and amplified...

...MDH). The MCC is a national group of educational leaders providing research opportunities to their students...MDH and, if indicated on the plasmid map, a TEV cleavage site is present between the 6xHis-tag and the ... region of MDH. For detailed clone information, please see the MDH Protein Clone Information Sheet, available... CUREs Community (Link opens in a new window) — Learn how to get started with a CURE, explore MDH projects...projects, find teaching resources and protocols, learn about regional hubs, access a teaching mentor, and...window) External Collaboration Results in Student Learning Gains and Positive STEM Attitudes in CUREs. Callahan...undergraduate research experiences (CURE) impacts student learning and attitudinal outcomes: A study of the Malate...

... 154948 CaMKII-somBiPOLES-mCerulean CamKII BiPOLES (soma-targeted) mCerulean Constitutive 5, 9, rg* Wiegert...154951 hSyn-DIO-somBiPOLES-mCerulean Syn BiPOLES (soma-targeted) mCerulean Cre dependent 5, 9, rg* Wiegert...Looking For? Please let us know what you would like to see available in viral format. Please note this does...Constitutive Serotype AAV1 AAV2 AAV5 AAV8 AAV9 AAVrg PHPeB Clear Filters ID Name Promoter Opsin Variant Tag Activity...available items found to match all requested criteria. Please consider using fewer filtering options. Browse ...

...-mediated nucleic acid cleavage. We have harnessed the type II CRISPR nuclease system to facilitate genome...nterspaced S hort P alindromic R epeats) is a microbial nuclease system involved in defense against invading phages...Nature Protocols 2013). For cloning information, please view: Zhang Lab General Cloning Protocol 237.2 ...by co-expressing the S. pyogenes Cas9 (SpCas9) nuclease along with the guide RNA. For most applications...gain-of-function screening. For more information, please visit the Addgene SAM library page . Targeted gene... directed repair-mediated Kras G12D mutations, leading to macroscopic tumors of adenocarcinoma pathology... DNA targeting specificity of RNA-guided Cas9 nucleases. Hsu PD, Scott DA, Weinstein JA, Ran FA, Konermann...

...Luciferase catalyzes the oxidation of a substrate, leading to the emission of photons. The robust signal, ...sensitivity, wide dynamic range, and resistance to photobleaching has made luciferase a common choice in assays...have made it popular with scientists since its release. Luciferase-containing plasmids are commonly used...applications. Browse the Collection Highlights section to learn more. Highlights of Luciferase Tools in our Collection...artificial luciferase desined de novo using deep-learning approaches. Split-Luciferase and Protein-Protein...luciferase. Plasmids are Gateway cloning compatible. Learn more about DULIP in our 27 Hot Plasmids from 2016...linker to the circularly permuted Venus fluorophore. Learn more about pHlash sensor in our 28 Hot Plasmid Technologies...

...protein function results in Rett syndrome is not clear, however, the ability to bind to methylated DNA ...ranscriptional R epressor D omain (TRD) the TRD contains a Nuclear receptor Co-Repressor 1/Silencing Mediator of Retinoic...receptor (NCoR/SMRT) interacting domain (NID) a nuclear localization signal (not essential for localization...RSRT cell line collection (186.8 KB) . To order, please email [email protected] The following cell lines are...information on generating iPSCs using plasmids, please see Addgene's Plasmids for Stem Cell Research page...precise editing of mutations in the MECP2 gene. Please refer to Addgene's CRISPR Guide for a general introduction...your plasmids! To get started with your deposit, please click here ....