pJSC281 - Bacterial expression plasmid for SpCas9 Cluster 1 (HypaCas9), REC2 FRET variant
(Plasmid
#101230)
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PurposeBacterial expression plasmid for SpCas9 Cluster 1 (HypaCas9), REC2 FRET variant
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Depositing Labs
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Publication
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Sequence Information
Ordering
This material is available to academics and nonprofits only.
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 101230 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 | |
Backbone
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Vector backbonepCT10
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Backbone manufacturerDoudna lab
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Vector typeBacterial 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 nameSpCas9 variant C80S/C574S/E60C/D273C/N692A/M694A/Q695A/H698A
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Alt nameStreptococcus pyogenes Cas9
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SpeciesStreptococcus pyogenes
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MutationC80S, C574S, E60C, D273C, N692A, M694A, Q695A and H698A
- Promoter T7
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Tags
/ Fusion Proteins
- 10x His (N terminal on backbone)
- MBP (N terminal on backbone)
- TEV site (N terminal on backbone)
Cloning Information
- Cloning method Gibson Cloning
- 5′ sequencing primer T7
- 3′ sequencing primer T7-term (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
Please visit https://www.biorxiv.org/content/early/2017/08/12/160036 for bioRxiv preprint.
How to cite this plasmid
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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:
pJSC281 - Bacterial expression plasmid for SpCas9 Cluster 1 (HypaCas9), REC2 FRET variant was a gift from Jennifer Doudna & Keith Joung (Addgene plasmid # 101230 ; http://n2t.net/addgene:101230 ; RRID:Addgene_101230) -
For your References section:
Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Chen JS, Dagdas YS, Kleinstiver BP, Welch MM, Sousa AA, Harrington LB, Sternberg SH, Joung JK, Yildiz A, Doudna JA. Nature. 2017 Sep 20. doi: 10.1038/nature24268. 10.1038/nature24268 PubMed 28931002
