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We narrowed to 62 results for: c-Myc-promoter

Showing: 21 - 40 of 62 results

  1. Plasmids 101: E. coli Strains for Protein Expression

    Type
    Blog Post
    ...hybrid promoter. Although the DE3/T7 RNAP system works well for most experiments, the lac promoter can “...expression  m15 pREP4*, ** Kanamycin (pREP4) Cis-repression of the E. coli T5 promoter (found on vectors such...expression from high-copy number plasmids and powerful promoters will greatly exceed that of any native host protein... accomodate high protein loads using inducible promoter systems (which will be discussed later). In addition...above, many expression plasmids utilize inducible promoters, which are 'inactive' until an inducer such as...the culture to make protein.  The DE3 lysogen/T7 promoter combination is the most popular induction system...available to transcribe the gene of interest from a T7 promoter on the plasmid. Many commercial strains carry ...

  2. Your Top Requested Plasmid in 2016!

    Type
    Blog Post
    ...pyogenes (Sp) Cas9 from a Cbh promoter. Cbh is a modified version of CAG promoter that the Zhang lab has tested...imporant is that, upstream of the Cbh promoter, this plasmid has a U6 promoter followed by BbsI restriction sites...site). Finally, in addition to the 2A peptide on its C-terminus, SpCas9 is also fused to a 3X FLAG tag at...SpCas9 is fused to the puromycin resistance gene through a 2A peptide. The puromycin resistance gene enables...causes the ribosome to translate Cas9-2A and the Puromycin resistance gene as separate peptides. PX459 This...predecessor, which had a point mutation in the puromycin resistance gene that made it less effective in...

  3. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    ...from its own promoter. But that doesn’t mean you have to clone and transfect multiple promoter-gRNA constructs...arrays (e.g. for spacers A, B, and C, you could obtain arrays A, B, C, AB, AC, BC, and ABC). Nielsen Lab...can I express more than one gRNA from a single promoter using a plasmid like pX330? Unfortunately, the...of pX330, contains humanized wtCas9 and two U6 promoters. To use this plasmid, you simply order oligonucleotides...contain Type IIS restriction sites flanking the promoter-gRNA construct, but with different sequences adjacent...contain Type IIS restriction sites that flank the promoter-gRNA constructs. When these plasmids are digested...downstream of the 7SK, human U6, mouse U6, or human H1 promoters. If you express fewer than four gRNAs, you’ll ...

  4. Neurodegeneration Plasmid Collection

    Type
    Collection
    ...pCMVTNT PINK1 N-myc PINK1 Myc CMV Parkinson's Mark Cookson 13314 pCMVTNT PINK1 C-myc PINK1 Myc CMV Parkinson's...Ultra-Exon1Q23-Myc-A HTT Myc UbC Huntington's Baoji Xu 110487 Ultra-Exon1Q145-Myc-A HTT Myc UbC Huntington's...pUltra-Exon1Q23-Myc-B HTT Myc UbC Huntington's Baoji Xu 110489 pUltra-Exon1Q145-Myc-B HTT Myc UbC Huntington's...Strep-Lrrk2-Myc LRRK2 Myc, Strep CMV Parkinson's Maik Hintze 161583 pPuro3.1(+)_Strep-Lrrk2-Myc LRRK2 Myc, Strep...LRRK2-WT LRRK2 His, Myc CMV Parkinson's Ted Dawson 17612 pRK5-Myc-Parkin PRKN Myc CMV Parkinson's Ted ... pGW1-Myc-DJ1-WT PARK7 V5 CMV Parkinson's Mark Cookson 29349 pGW1-Myc-DJ1-L166P/K130R PARK7 Myc CMV Parkinson's...D1994S)-Myc LRRK2 Myc, Strep CMV Parkinson's Maik Hintze 161585 pPuro3.1(+)_Strep-Lrrk2(G2019S)-Myc LRRK2...

  5. Hot Plasmids Spring 2024

    Type
    Blog Post
    ...screening of human promoters, the Zhang Lab found a short fragment of human IBA1 promoter (hIBA1a, 466 bp...small RNA-binding exonuclease protection factor) promotes editing (Yan, J et al., 2024). They developed ... expression from ssAAV5s with hIBA1 or hIBA1a promoters in mice. B) Quantifications showing high microglia...a CC-BY-NC-ND license.  Integrating hIBA1a promoter into AAV vectors, either alone or in combination...specificity of GFP expression for the indicated ssAAV5s. C) Quantifications showing high microglia transduction... microglia-targeting AAV plasmids here! Serrano, C., et al. (2023). Simple and Highly Specific Targeting... vectors for CROPseq-multi using selection by Puromycin, Zeocin®, or nuclear expression of BFP2.   ...

  6. Tips for Screening with Yeast Two Hybrid Systems

    Type
    Blog Post
    ...El-Bakkoury, M., Hamacher, T., Cappellaro, C., Vilarino, C., Fleischer, C., Ellerbrok, H., Kamuzinzi, R., Ledent...contains a reporter gene cloned downstream of a promoter containing the target sequence of the DBD. The...as the “prey”) in close enough proximity to the promoter of the reporter gene to activate expression (Figure...organisms, including bacteria, alternative fungi (C. albicans, pC2HB) (7) and mammalian cells. If the ... prey libraries can be screened using both N- and C-terminal fusions of these proteins. This way both ...Two hybrid systems were developed in Saccharomyces cerevisiae in 1989 and are still used extensively ...

  7. CRISPR/Cas9 FAQs Answered!

    Type
    Blog Post
    ...sequence into a backbone that uses the human U6 promoter to drive expression, is it necessary to add a ... start of my target sequence? A3: The human U6 promoter prefers a 'G' at the transcription start site ...of the crRNA) is also expressed from a separate promoter. In the pre-processing form, the crRNA contains...10.1126/science.1231143  Elliott, B., Richardson, C., Winderbaum, J., Nickoloff, J. A., & Jasin, M. (1998....1128/mcb.18.1.93  Ran, F. A., Hsu, P. D., Lin, C., Gootenberg, J. S., Konermann, S., Trevino, A. E....j.cell.2011.06.019  Wang, H., Yang, H., Shivalila, C. S., Dawlaty, M. M., Cheng, A. W., Zhang, F., & Jaenisch...j.cell.2013.04.025  Yang, H., Wang, H., Shivalila, C. S., Cheng, A. W., Shi, L., & Jaenisch, R. (2013)....

  8. Making CRISPR Plasmids Using Fragmid

    Type
    Blog Post
    ...reporters, Cas proteins, guide cassettes, promoters, N’ and/or C’ terminus, and selection markers. Need ...components, such as your CRISPR enzyme or Pol 2 promoter. The options available in each class, or module...Griffith, A. L., Szegletes, Z. M., Wen, B., Kraus, C., Miller, N. W., Steger, R. J., Escude Velasco, B....Plasmid pRDB_052 (for stable fly cell lines; Hygromycin resistance gene; for genome integration by spontaneous...

  9. Bacterial Expression Systems

    Type
    Collection
    ...pCHERRY3 Promoter activity Fluorescence (mCherry) Mycobacterium sp. Tanya Parish 24657 pASTA3 Promoter activity...coli strain. Addgene Blog Inducible Promoters Repressible Promoters Synthetic Photobiology: Optogenetics...pRU1701 Promoter activity Fluorescence (GFP+) Gram-negative bacteria Philip Poole 14460 pOT2 Promoter activity...peptides include: Epitope tags: 6xHis, Flag, Strep II, c-Myc, HA, V5, GST Solubility tags: MBP, SUMO, TrxA, Mocr...Acetamidase promoter Acetamide Escherichia coli , Mycobacterium smegmatis Matthias Wilmanns 84692 pMyC-kan Acetamidase... Acetamidase promoter Acetamide Escherichia coli , Mycobacterium smegmatis Matthias Wilmanns 84689 pMyBADC-kan...Fluorescence (tdTomato) Mycobacterium sp. Tanya Parish 24658 pCHARGE3 Promoter activity Fluorescence (...

  10. Rewiring Metabolic Circuitry with CRISPR RNA Scaffolds [Video]

    Type
    Blog Post
    ...limited by the lack of well characterized inducible promoters. Far removed from the bio-industry’s vision of... the same dCas9 protein. By using an inducible promoter to modulate dCas9 expression, multiple scRNA transcriptional...on designing and using RNA scaffolds: Delebecque, C. J. et al. Designing and using RNA scaffolds to assemble... various metabolites in Escherichia coli or Saccharomyces cerevisiae with few techniques that are easily...

  11. Pathways Over Time Plasmids Engage Students in Functional Genomics Research

    Type
    Blog Post
    ... by the GAL1 promoter. The cloning procedure adds both V5 and His6 epitope tags to the C-termini of Met...grow in the absence of methionine. Since the GAL1 promoter controls MET/Met gene expression, complementation...methionine synthesis (5). The budding yeast, Saccharomyces. cerevisiae, makes a great reference organism... or not proteins from the fission yeast, Schizosaccharomyces pombe, have the same function as their counterparts...yeast for budding geneticists: A primer on the Saccharomyces cerevisiae model system. Genetics 197, 33-48...yeast for young geneticists: A primer on the Schizosaccharomyces pombe model system. Genetics 201, 402-423...Surdin-Kerjan, Y. Metabolism of sulfur amino acids in Saccharomyces cerevisiae. Microbiol Mol Biol Rev 61, 503-532...

  12. Simple CRISPR-based Epigenetic Editing: dCas9-directed DNA Demethylation

    Type
    Blog Post
    ...02708-5 Hrit J, Goodrich L, Li C, et al. (2018) OGT binds a conserved C-terminal domain of TET1 to regulate...functional role of DNA demethylation at specific promoters by targeted steric blockage of DNA methyltransferase...89–92. https://doi.org/10.1038/301089a0. Galonska C, Charlton J, Mattei AL et al (2018). Genome-wide tracking...https://doi.org/10.7554/eLife.34870.  Kaas GA, Zhong C, Eason DE, Ross DL, Vachhani RV, Ming G-L, King JR...lentiviral backbones expressing blasticidin or puromycin resistance genes.  It’s that simple! Good luck...

  13. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    ...either the Thy1 or CAG promoter, while Brainbow AAV is under control of the EF1a promoter. Livet et al., ...insert modules (plant promoter, N-terminal tag, coding sequence of the gene of interest, C-terminal tag, plant...known as SpyLigase and is a protein domain that promotes the formation of an isopeptide bond between 2 ...insert fragments of DNA containing basic parts (promoters, UTRs, coding sequences, terminators, etc) into... creating a single transcriptional unit (Ex: a promoter, 5’UTR, coding region, and terminator). Next, ...the mitochondrial targeting sequence of cytochrome c oxidase subunit VIII, so that they can be imported...pCoofy series of plasmids contain a variety of N- and C-terminal tags (including His, S-tag, OneStrep, CBP...

  14. Hot Plasmids and Viral Preps - March 2021

    Type
    Blog Post
    ...inhibitor response 1 (TIR1) expressed under a desired promoter with a specific spatiotemporal expression pattern...interactions New auxin inducible degron system for C. elegans New CRISPR plasmids New items from the viral...tool-kit for the auxin inducible degron (AID) system in C. elegans By Alyssa Cecchetelli The auxin inducible... degron (AID) system has been extensively used in C. elegans since 2015 to conditionally degrade proteins...repression, and editing with a single Cas9-VPR in Saccharomyces cerevisiae. To control Cas9 activity and influence...

  15. Harnessing TnpB for Genome Editing: A Compact and Versatile Tool for Your Research

    Type
    Blog Post
    ...choice of promoter for ωRNA (reRNA) expression (see Figure 1). Both plasmids contain a hygromycin resistance...Maher, M. F., Nasti, R. A., Vollbrecht, M., Starker, C. G., Clark, M. D., & Voytas, D. F. (2019). Plant gene...0337-2  Molla, K. A., Sretenovic, S., Bansal, K. C., & Qi, Y. (2021). Precise plant genome editing using...Gao, Q., Liu, S., Zhang, X., Hu, W., Wang, Y., Han, C., Li, D., & Wang, X. (2025). Transgene- and tissue...resistance gene as the plant-selectable marker and a kanamycin resistance gene for bacterial selection. The process...

  16. Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View

    Type
    Blog Post
    ...optimum of 30 °C and is therefore inefficient in mammalian cells (usually grown at 37 °C). Smart molecular...localization sequence) and expressing it from a CAGGs promoter. However FLPe is still less efficient than Cre...identification of FLPe which has a temperature optimum of 37 °C (Buchholz, Angrand, and Stewart 1998). FLPe’s performance... receptor (ER) ligand binding domain (LDB) to the C-terminus of FLP or Cre (pCAG-CreERT2 #14797). These...Sadowski. 1985. “The FLP Recombinase of the Saccharomyces Cerevisiae 2 Micron Plasmid Attaches Covalently...

  17. Plasmids 101: Broad Host Range Plasmids

    Type
    Blog Post
    ...Loh, P. C., & Wong, H. L. (2023). Development of constitutive and IPTG-inducible integron promoter-based...resistance cassettes (Prior et al. 2010), inducible promoters with different degrees of stringency and inducibility...Gluconacetobacter xylinus, Haemophilus influenzae, Hypomycrobium X, Legionella pneumophila, Methylophilus methyltrophus...calcoaceticus, Actinobacillus pleuropneumoniae, Actinomyces naeslundii, A. viscosus, Aerobacter aerogenes...Methylophilus methylotrophus, Moraxella spp., Mycobacterium aurum, M. smegmatis, Paracoccus denitrificans...spheroides, Salmonella spp., Serratia marcescens, Streptomyces lividans, Synechococcus spp., Thiobacillus ferrooxidans...

  19. Getting the Most from Your Lentiviral Transduction

    Type
    Blog Post
    ...enough to prevent confluency but high enough to promote cell growth. Overgrown cultures stimulate quiescence... for less than a day, lentivirus can be kept at 4°C. For long-term storage, viral preps should be divided...divided into single-use aliquots, and stored at -80°C. Some reports suggest rapidly freezing virus in a ... labs and hinders experiments is mycoplasma contamination. Mycoplasma contamination has been shown to ...routinely test for mycoplasma contamination. There are several commercially available mycoplasma detection kits...larger microbes such as yeast, fungi, or bacteria, mycoplasma can be extremely hard to detect with levels reaching...cells per ml before the media becomes cloudy. Mycoplasma compete with host cells for nutrients and can...

  20. Multiple Plasmids at a Low Price: Kits from Addgene

    Type
    Blog Post
    ... vectors within this kit contain a variety of promoters to facilitate gene expression in bacteria, yeast...plasmids within it contain a variety of different promoters followed by leader sequences (encoding small leader...different genes was predictable based solely on the promoters and Shine-Delgarno sequences used in this so-called... easy for a researcher to quickly generate N- and C-terminal epitope tag fusions to a protein of interest...et al. "Adaptation of Enterococcus faecalis to daptomycin reveals an ordered progression to resistance....
Showing: 21 - 40 of 62 results