...became known as SNAP-tag. There is also a fast-reacting version of SNAP-tag called SNAPf. eDHFR: The Virginia...Characterization of the Self-Labeling Protein Tags HaloTag7, SNAP-tag, and CLIP-tag. Biochemistry, 60(33), 2560–2575...than short peptide tags like tetracysteine (Jing & Cornish, 2011). HaloTag and SNAP-tag are the most popular...application, SNAP-tag, CLIP-tag (both 182 aa), and eDHFR (159 aa) are smaller than HaloTag (295 aa) – for...on resorufin). SNAP-tag: In 2003, Kai Johnsson’s lab developed a self-labeling fusion tag based on the ...Although based on the same original enzyme, SNAP-tag and CLIP-tag show good specificity for their own ligands...genetic tag on the protein of interest. Table 1: Summary of popular self-labeling tag systems Tag Size...

...resistance genes, selectable markers, promoters, origins, tags, and certain ORFs. Their sophisticated detection...powered by GSL Biotech's SnapGene Server Software. With the backing of SnapGene’s sequence viewer software... Overview of SnapGene powered plasmid maps at Addgene  Three factors make our SnapGene powered plasmid...the features you need quickly. SnapGene’s Feature Library - SnapGene’s software seamlessly identifies ...example below. Our old map is on the left while the SnapGene powered map is on the right (click here to see...t detected by our in-house software. In fact, SnapGene’s feature library includes nearly 3x as many features...identify. Improved Restriction Enzyme Handling - SnapGene’s software positions restriction enzyme annotations...

...markers, tags, origins of replication, and fluorescent proteins that are already included in Snapgene’s extensive...through our Snapgene powered maps and sequence analysis tools.     The main ways we use SnapGene for plasmid...version of Snapgene to visualize the sequence and look for features. The first thing that the Snapgene software...software as the desktop version of Snapgene. When you download a Snapgene file directly from our website,...website, you can open it with either the Snapgene viewer or Snapgene Desktop programs to analyze the sequence... plasmids. We’ve also talked about how our new Snapgene generated maps provide improved feature detection...with an easy to use interface. We regularly use Snapgene for our quality control process because of its...

... who uses SnapGene or the free SnapGene Viewer companion product. If you don’t use SnapGene yet, you can...guest bloggers Aline and Benjamin Glick from SnapGene. SnapGene was created to meet a need. While there were...approach is SnapGene’s algorithm for detecting common features. This algorithm enables one of SnapGene’s most... us at snapgene.com. Many thanks to our guest bloggers Aline and Benjamin Glick from SnapGene!  Ben Glick...Addgene Blog Read about our SnapGene powered plasmid maps Learn how we use SnapGene during the plasmid quality...properties of the DNA molecules they were using. SnapGene was created to alleviate these problems through...make specialized controls available when needed. SnapGene has been engineered to be easy and enjoyable to...

...wintertime festivities! Help us spread some cheer by snapping a photo of you and your labmates celebrating the... the holidays and tweeting it with the tags #DeckTheLab and @Addgene. Christmas, Hanukkah, ugly sweaters...you name it, we want to see it.      All photos tagged with #DeckTheLab throughout the month of December...

... Why #DecktheLab? Help us spread some cheer by snapping a photo of you and your labmates celebrating the... the holidays and tweeting it with the tags #DeckTheLab and @Addgene. We’d love to see how you’re decorating...Festivus— you name it, we want to see it! All photos tagged with #DeckTheLab throughout the month of December...

...other fluorescent proteins and tags including mCherry, HaloTag and SNAP-tag for multi-color imaging. Find...observed colocalization of GFP tagged HA frankenbody with an mCherry, 4xHA tagged histone protein (H2B) in ...could also bind a 1xHA tagged histone protein as well as a mitochondrial protein tagged with spaghetti monster... the 10xHA tag increased the fluorescent signal to noise ratio five fold over the 1xHA tag.  Lastly, the...use GFP to image nascent proteins. In addition GFP tags are large, dim, and permanently attached to a protein...limitations, scientists have co-opted antibodies for tagging purposes. Different antibodies bind different protein... called epitopes and scientists can use these to tag their proteins of interest. Antibody based probes...

... with containing chemical tags such as the SNAP-tag, Halo-tag, and CLIP-tag (Kohl et al., 2014) (Addgene... et al. "Ultrafast tissue staining with chemical tags." Proceedings of the National Academy of Sciences111.36... circuits with fluorescent proteins and chemical tags." eLife 7 (2018): e40350. PubMed PMID: 30454553....

...frankebodies includes mEGFP, mRuby2, iRFP713, SNAP-tag, and HaloTag fusions, while the anti-HA frankenbodies... small epitope tag (FLAG is eight amino acids, HA is nine), multiple repeats of the tag can recruit multiple...b) Schematic of anti-FLAG-FB-GFP binding to FLAG-tag H2B for nuclear localization. (c) Applications of...blotting, and more. Given the popularity of FLAG and HA tags, you may not even need to clone new constructs to.... The proteins, which are purified via histidine tags, are optimized for SDS-PAGE gels and can be detected...frankenbodies include mEGFP, mCherry, and HaloTag. Figure 1: (a) Anti-FLAG frankenbody (FB) construction from...editing within a single expression cassette is advantageous to Cas9’s larger expression requirements (Figure...

...T2A-TurboGFP-PEST), and small epitope tags (HA, Myc, Strep tag II, AviTag, HaloTag, SpyTag). The Foerstemann lab developed...types of fluorescent (EGFP, tagRFP, mCherry) and nonfluorescent (Halo, SNAP) tags, a selectable marker (floxed... scarless tag insertion.   In addition to the obvious advantages SapTrap offers for tagging a single genetic...between the tag and homology arms). Digestion of the donor plasmids with SapI allows the tag, selectable... inserting a tag at multiple target sites. SapTrap vectors can be easily modified to tag hundreds of genes...insertion tagging), developed by the Hornung Lab, allows users to easily create C-terminal tag fusions ...endogenous protein tagging, please visit our website CRISPR/Cas Plasmids - Protein Tagging. Beyond simply...

...concept include: Self-labeling tags (e.g., SNAP-tag, HaloTag, TMP-tag) – These widely used systems consist...strategies have two parts: (1) a genetically encoded “tag” expressed as a fusion with your favorite protein...fluorophore-contianing “ligand” that binds to the tag. Like most good ideas in biological imaging, the ...genetically encoded tetracysteine (Cys4) peptide tag could be used to label proteins in cells (Figure ...consist of a genetically encoded enzyme variant tag that reacts specifically and irreversibly with a small... attachment of a fluorophore ligand to a peptide tag (Figure 1c). Click chemistry (e.g., transcyclooctene–tetrazine...paclitaxel. Unlike other systems, no genetically encoded tag is required as the small-molecule binding motif targets...

...C-terminal fusion of a tetracysteine tag. In addition, since these tags are so small, one can insert them... lure the ribosome into inserting a fluorophore-tagged amino acid at a stop codon (often amber i.e. UAG...termination at stop codons, expression levels of these tagged proteins are often very low. In addition, a major...which can be carried out using Addgene plasmids. His-tag labeling reagents You may already have a FRET acceptor-binding...: you just don’t know it yet! The poly-histidine tag often used for protein purification is a reliable...for protein purification: the NTA binds to the His tag on your protein and the coupled Cy fluorophore acts...well as by extending the length of the histidine tag from 6 to 10 residues. While these Cy/NTA conjugates...

... would be SNAP-tag® and HaloTag®. These consist of a genetically encoded enzyme variant tag that covalently...several advantages to using these self-labeling proteins over traditional fluorescent protein tags, such...endogenous target, and thus no genetically encoded tag is required. These dyes can also be attached to an...ligands have more flexibility due to the affinity tag they typically bind to. Stability of fluorescence...experimental labeling. Size is also a factor. Bulkier tags, such as GFP, may interfere with protein expression...fluorescent experiments, such as using fluorescently tagged ligands or intermolecular FRET for ligand-receptor...should possess high specificity to ensure accurate tagging. In Fluorescence Resonance Energy Transfer (FRET...

... storage, make > 25 μl aliquots in 0.5 ml tubes, snap freeze in liquid nitrogen or a dry ice/ethanol bath...them. There are suspected cases of insertional mutagenesis in humans (1) and AAVs are non-enveloped viruses... Russell, David W. "AAV vectors, insertional mutagenesis, and cancer." Molecular Therapy 15.10 (2007):...

...identify as one.  But some time around 10th grade, I snapped out of the phase of my life that had me listening...the first place? Let’s think about some of the advantages of team-based work: Nobody knows everything anymore...

...and gram negative bacteria periplasm. Efforts to tag endogenous genes for proteins in these compartments...Munro, Sean, and H. R. Pelham. "Use of peptide tagging to detect proteins expressed from cloned genes:...Avaliable at Addgene Read Our Blog Post on Protein Tags Read Other Posts on Fluorescent Proteins Browse ...our guest bloggers Erik L. Snapp and Lindsey M. Costantini! Erik Lee Snapp received his PhD in Molecular... post was contributed by guest bloggers Erik L. Snapp and Lindsey M. Costantini. "You underestimate the...FP-fusion proteins functionally relative to the untagged protein of interest. To avoid the accumulation...research was conducted in the laboratory of Erik Lee Snapp and focused on optimizing fluorescent proteins for...

... Replace the biotinalatable tag on dCas9 with a BioTAP-tag. The BioTAP-tag is a 69 aa long biotinylation... lentiviruses delivering 1) a BioTAP-tagged dCas9 and an eGFP tag, and 2) gRNAs Number of loci targeted...normally expressed in eukaryotic cells. Using this tag reduces the number of CAPTURE components that need...from three to two. Lentiviral delivery of BioTAP-tagged dCas9 and gRNAs. Using lentiviral delivery of the...system Stable cell line expressing 1) FLAG-biotin-tagged dCas9, 2) BirA, and 3) one or more gRNAs Two separate...et al., 2018). Think of 3C methods as taking a snapshot of chromatin looping with the photo “developed...

...solution or something in between, you are using a tag with a serious drawback. The standard EGFP plasmid... of a POI. As an investigator, you want a fusion tag to be inert, to not produce artifacts in your experiments...Many thanks to our guest blogger Erik L. Snapp! Erik Lee Snapp received his PhD in Molecular Microbiology...This post was contributed by guest blogger Erik L. Snapp. Stop using EGFP/GFP for fusion proteins! Despite...oligomers such as DsRed or high affinity FPs such as TagRFP. Problem 3 can be solved, in most cases, by using...distance runner, gardner, and cook.   References 1. Snapp, E.L., et al., Formation of stacked ER cisternae... Resources on the Addgene Blog Read Professor Snapp's Post on mOX FPs Find the Best Fluorescent Protein...

...into a vector with a large tag at the N-terminus, but you don’t have the tag already included in the vector...you can insert both the gene of interest and the tag sequences into the vector in one step without scars...In this blog post, I will go over some advantages, disadvantages, and examples of how scientists are using... common, and for good reason. They offer many advantages over the traditional restriction enzyme cloning...thereof) nor a serious time commitment. Another advantage is that this process makes it easy to generate...primer melting temperature greater than 48℃. Both Snapgene and NEB have tools that help you design primers...video gives a helpful demonstration of how to use Snapgene’s program to design primers for Gibson Assembly...

...plasmid.     Confirming tags and fusion proteins Finally, we confirm promoters, tags, fusion proteins, and...aligning to NCBI entry or using BLAST.   Confirming tags and fusion proteins. We’ll break down each of these...selectable markers by detecting common features using Snapgene. If we find information that differs from what...