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  1. Plasmids 101: The Promoter Region – Let's Go!

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    Blog Post
    ... trying to pick your perfect promoter! Eukaryotic Promoters   Promoter   Primarily used for RNA     .... In practice, the term "promoter" describes the combination of the promoter (RNA polymerase binding site...recognize and bind to specific promoter elements. This means that the promoter present in your plasmid backbone...RNAP II promoter, whereas small RNAs (such as shRNA) are transcribed from the RNAP III promoters. This ...post on viral vector parts. Promoter specificity Aside from choosing a promoter based on type of RNA transcript...cell types or organisms, promoters must be similarly variable. Bacterial promoters only work in prokaryotic...require unique promoters and there is very little crossover. Generally speaking, promoters in bacteria are...
  2. New Acoustic Reporter Genes: Ultrasound Imaging of Gene Expression

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    Blog Post
    ...constitutively-expressed mARG plasmids driven by CMV promoters. Labs that are new to the mARG expression are...cotransfect a mixture of 1248 ng of CMV-gvpA-IRES-BFP plasmid and 954 ng CMV-gvpNJKFGWV plasmid into 70%-confluent...expression systems in E. coli. The arabinose-inducible promoter pBAD and a low-copy number origin of replication...integratable cassettes, both with doxycycline inducible promoters (Fig. 2a). The first cassette contains the major...
  3. AAV Q&A with Tim Miles

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    Blog Post
    ...efficient AAV prep for a 4.2 kB insert with the CMV promoter using the AAV-BR1 capsid? A: It is our understanding... ~4.7kB.  If your insert alone is 4.2 kB and CMV promoter+enhancer is 0.5 kB, then you would be oversized...
  4. Building and Validating Recombinant Antibodies: A Case Study

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    Blog Post
    ...light chain, each from a constitutively active CMV promoter. Constructs were then verified by both diagnostic...green) and heavy (blue) chains as driven by two CMV promoters (orange). Figure from Andrews, et. al., Elife...
  5. Making CRISPR Plasmids Using Fragmid

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    Blog Post
    ...components, such as your CRISPR enzyme or Pol 2 promoter. The options available in each class, or module...out reporters, Cas proteins, guide cassettes, promoters, N’ and/or C’ terminus, and selection markers....lentiGuide-based) Mammalian Lentivirus pRDA_722 (CMV-driven; lentiCRISPRv2-based; preferred over 512) ...) Mammalian Lentivirus pRDA_789 (CMV-driven; CROPseq) Mammalian AAV pRDA_889 (ITRs from...
  6. Plasmids 101: Inducible Promoters

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    Blog Post
    ...will be produced. Many common promoters. like CMV, EF1A, and SV40 promoters, are always active and thus ... on repressible promoters. How are inducible promoters regulated? Inducible promoters can be regulated...commonly used prokaryotic promoters is the negative inducible pLac promoter. This promoter requires removal of...steroid regulated promoters commonly used in plant research. Promoter Subtype Promoter Example Activator...Promoters control the binding of RNA polymerase and transcription factors. Since the promoter region ...of a promoter. Below, you’ll find a short description of these three types of inducible promoters, and...Many of these promoter systems are available at Addgene! Chemically inducible promoters Chemically regulated...
  7. Choosing Your Perfect Empty Backbone

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    Blog Post
    ...pcDNA3, comes from the pcDNA series. It has the CMV promoter for mammalian expression, a multiple cloning...yeast expression vectors with various markers and promoters. If you need to visualize YGOI on a western blot...vectors carrying several fusion proteins. Pick your promoter, selection cassette and visualization vehicle ...fly genome and are under the Gal4 inducible UAS promoter. Other vectors can be expressed in fly culture...culture cells like S2 and carry the metallothionein promoter (e.g. pMT-puro). Worm vectors There are several... following the expression pattern of a defined promoter region, ectopically expressing YGOI in a pattern...
  8. 28 Hot Plasmid Technologies from 2015

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    Blog Post
    ...vector containing a promoter for constitutive (CMV, SV40, SFFV) or inducible (CMV-TO) cDNA expression,...fluorescent proteins, 8 constitutive promoters, 2 includible promoters, 3 polyA terminators and various pieces...sequences. Several pre-constructed promoter-selection marker or promoter-inducible expression related vectors...plasmids: Luc (intronless; CMV-LUC2CP/ARE and Luc-I (intron-containing; CMV-LUC2CP/intron/ARE). The Luc-I...all chemically inducible promoters; 3) improving the strength of the promoters used; and 4) optimizing ...ideal promoter/location combination. However, other constructs which contain different promoters and/or...to study the human GM-CSF promoter and enhancer, a finely regulated promoter controlled by a mixture of...
  9. Plasmids 101: Protein Expression

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    Blog Post
    ...designed for mammalian expression utilize viral promoters (SV40, CMV, and RSV) for robust expression post-transfection...the use of constitutive and inducible promoters. Inducible promoters are extremely useful if a desired protein... of the galactose inducible promoter (GAL). Other commonly used promoters include the phosphate and copper... Blog Learn about inducible promoters Learn about repressible promoters Check out protein tags Resources... of these vectors is controlled by a different promoter resulting in different levels of expression from... vectors, pET, under the control of the T7 lac promoter and induced by lactose, provides the highest level...copper inducible PHO5 and CUP1 promoters respectively. Yeast cells are grown in well-defined media and...
  10. Introducing Addgene's New Search - Find Plasmids More Easily For Your Research!

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    Blog Post
    ...plasmid to express ubiquitin under control of a CMV promoter. Starting with two search terms we see there...scientists Viral prep filter categories Promoters: Select the best promoter for your target cell type Serotype...world, it can be hard to find your favorite gene, promoter, serotype, or a myriad of other things. That’s...
  11. Plasmids 101: Repressible Promoters

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    Blog Post
    ... are repressible promoters regulated? Like inducible promoters, repressible promoters can be regulated...Promoters control the binding of RNA polymerase and transcription factors. Since the promoter region ... will be produced. Many commonly-used promoters, such as T7, CMV, EF1A, and SV40, are always active and...natural and engineered repressible promoters. Chemically repressible promoters Tet-Off In its native context... pADH1 displays low promoter activity (~20% activity of the strong yeast promoter pTEF). As ethanol accumulates...that promoter is active. In this way, you can interrogate the activity of uncharacterized promoters. Similarly... QUAS promoter, and the QF transcription factor cannot drive transcription from the UAS promoter. This...
  12. MXS Chaining

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    Blog Post
    ...(Table 1). Each construct was flanked with a CMV promoter (to drive high-level expression) and a polyA... may differ (e.g., a set of various mammalian promoters), the same cloning procedure can be used to incorporate... in this method include fluorescent proteins, promoters, enhancers, polyadenylation signals, inducible...multiple identical polyadenylation signals or promoters Gibson et al., 2009 MXS-Chaining, BioBricks,...
  13. CRISPR 101: Ribonucleoprotein (RNP) Delivery

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    Blog Post
    ... cells where common eukaryotic promoters (such as CMV or EF1A promoters found in many CRISPR plasmids)...transfection or viral transduction     Appropriate promoters must be chosen for both Cas9 and gRNA expression...
  14. Your Lentiviral Plasmid FAQs Answered

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    Blog Post
    ...plasmids have a chimeric 5'LTR that includes a CMV or RSV promoter as well as a portion of the 5'LTR. Including...majority of transfer plasmids use the weak viral LTR promoter to drive expression of the gene of interest. Consequently...2nd-generation system because the wildtype 5'LTR promoter requires Tat to function. Third-generation plasmids...
  15. Multicolor Animals: Using Fluorescent Proteins to Understand Single Cell Behavior

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    Blog Post
    ...tandem with a single promoter, but only the gene directly following the promoter is expressed. Cre-mediated... As RFP and OFP are directly downstream of the promoter in Brainbow 1.0 and 1.1, respectively, cells will...Brainbow system are that it does not use ubiquitous promoters, meaning that not all cell types can be labeled...Brainbow was also used in zebrafish by injecting a CMV-Brainbow construct at the embryonic stage. This technique...placing Brainbow downstream of a neural-specific promoter, or the entire fish could be labeled by injecting...an RFP, CFP, and YFP downstream of a ubiquitous promoter. Zebrabow is an extremely versatile technology...
  16. Kazuhiro Oka Lentiviral Vectors

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    Collection
    ...from the CMV promoter pAdx-CMV-YFP 73348 Express YFP with an NLS from the CMV promoter pAdx-CMV-dsRed 73349...gene from the CMV promoter pCDH-CMV 72265 Express gene of interest from the CMV promoter pCDH-EF1 72266...from the CMV promoter pAdx-CMV-iCre-p2A-copGFP 73350 Expresses iCre and EGFP from the CMV promoter pAdx-CMV-iCre-P2A-tdTomato...EF1 promoter while pICPIS-CB contains the Chicken-beta actin promoter with CMV enhancer. pAdx-CMV-LacZ...Xgal/IPTG plate pAdx-CMV-copGFP 73346 Expresses copGFP from the CMV promoter pAdx-CMV-tdTomato 73347 Expresses...phosphoglycerate kinase I) promoter pCDH-CMV4 72284 Express gene of interest from the CMV promoter. WPRE has been ...truncated EF1 promoter pCDH-CMV-Nluc-P2A-copGFp-T2A-Puro 73037 Expresses Nluc and copGFP from the CMV promoter...
Showing: 1 - 20 of 54 results