CAG::PSAML141F:GlyR-IRES-GFP
(Plasmid
#32478)
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Depositing Lab
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Publication
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 32478 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepCAGGS- IRES-GFP
- Backbone size w/o insert (bp) 7100
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Modifications to backboneWPRE (Woodchuck hepatitis virus posttranscriptional regulatory element) before polyA sequence
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Vector typeMammalian Expression
Growth in Bacteria
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Bacterial Resistance(s)Ampicillin, 100 μg/mL
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Growth Temperature37°C
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Growth Strain(s)DH5alpha
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert namePSAML141F-GlyR
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SpeciesH. sapiens (human)
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Insert Size (bp)1295
- Promoter CAG
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Tag
/ Fusion Protein
- IRES-GFP (C terminal on backbone)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site NheI (not destroyed)
- 3′ cloning site NotI (not destroyed)
- 5′ sequencing primer CTGCTAACCATGTTCATGC
- 3′ sequencing primer GAAGACTCTTGCGTTTCTG (Common Sequencing Primers)
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
Depositor Comments
Note on the difference between PSAMY115F,L141F:GlyR and PSAML141F:GlyR silencers:
In Magnus et al, the PSAMY115F,L141F-GlyR channel was emphasized for silencing because this channel has the lowest acetylcholine responsiveness. However, PSAML141F-GlyR constructs, lacking the Y115F mutation, already have low ACh potency. The Y115F mutation also slightly increases the EC50 for the activator ligand, PSEM89S. Since PSAML141F-GlyR constructs already have very low acetylcholine potency, in many cases it may not be worth the slight reduction in PSEM potency. We have made both constructs available for researchers who may find that they have different requirements with respect to acetylcholine responsiveness.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
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For your Materials & Methods section:
CAG::PSAML141F:GlyR-IRES-GFP was a gift from Scott Sternson (Addgene plasmid # 32478 ; http://n2t.net/addgene:32478 ; RRID:Addgene_32478) -
For your References section:
Chemical and genetic engineering of selective ion channel-ligand interactions. Magnus CJ, Lee PH, Atasoy D, Su HH, Looger LL, Sternson SM. Science. 2011 Sep 2;333(6047):1292-6. 10.1126/science.1206606 PubMed 21885782