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PurposeHuman expression plasmid for SpCas9-VRQR-HF1 variant: CMV-T7-humanSpCas9-VRQR-HF1(N497A, R661A, Q695A, Q926A, D1135V, G1218R, R1335Q, T1337R)-NLS-3xFLAG
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 72252 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backboneJDS246
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Backbone manufacturerJoung lab (Addgene Plasmid # 43861)
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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 namemammalian codon-optimized Streptococcus pyogenes Cas9 VRQR-HF1(N497A/R661A/Q695A/Q926A/D1135V/G1218R/R1335Q/T1337R)-NLS-3xFlag
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SpeciesS. pyogenes
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MutationN497A, R661A, Q695A, Q926A, D1135V, G1218R, R1335Q, and T1337R
- Promoter CMV
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Tags
/ Fusion Proteins
- NLS (C terminal on insert)
- 3x FLAG (C terminal on insert)
Cloning Information
- Cloning method Unknown
- 5′ sequencing primer CMV-F
- 3′ sequencing primer BGH-rev (Common Sequencing Primers)
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.
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:
MSP2443 was a gift from Keith Joung (Addgene plasmid # 72252 ; http://n2t.net/addgene:72252 ; RRID:Addgene_72252) -
For your References section:
High-fidelity CRISPR-Cas9 nucleases with no detectable genome-wide off-target effects. Kleinstiver BP, Pattanayak V, Prew MS, Tsai SQ, Nguyen NT, Zheng Z, Keith Joung J. Nature. 2016 Jan 6. doi: 10.1038/nature16526. 10.1038/nature16526 PubMed 26735016