B-evoSpCas9-E526K
(Plasmid
#207365)
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PurposeExpression plasmid for human codon-optimized increased-fidelity (i.e. high-fidelity) SpCas9 variant
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 207365 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepX330-like (without U6-sgRNA coding sequence)
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Vector typeMammalian Expression, CRISPR
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 nameB-evoSpCas9-E526K
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SpeciesS. pyogenes
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Insert Size (bp)4251
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MutationM495V, Y515N, R661Q, amino acids 1005-1013 replaced with two glycine
- Promoter CBh
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Tag
/ Fusion Protein
- 3xFLAG (N terminal on insert)
Cloning Information
- Cloning method Gibson Cloning
Resource Information
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Supplemental Documents
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
Please visit https://doi.org/10.1101/2022.08.16.504097 for bioRxiv preprint
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:
B-evoSpCas9-E526K was a gift from Ervin Welker (Addgene plasmid # 207365 ; http://n2t.net/addgene:207365 ; RRID:Addgene_207365) -
For your References section:
A cleavage rule for selection of increased-fidelity SpCas9 variants with high efficiency and no detectable off-targets. Kulcsar PI, Talas A, Ligeti Z, Toth E, Rakvacs Z, Bartos Z, Krausz SL, Welker A, Vegi VL, Huszar K, Welker E. Nat Commun. 2023 Sep 16;14(1):5746. doi: 10.1038/s41467-023-41393-5. 10.1038/s41467-023-41393-5 PubMed 37717069