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PurposeLentiviral CRISPR-Cas9 delivery for SpCas9 and sgRNA. Coexpresses tagRFP via P2A cleavage site. EFS Promoter driven
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 57819 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepLKO.005
- Total vector size (bp) 11701
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Vector typeLentiviral, 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)Stbl3
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Copy numberHigh Copy
Gene/Insert 1
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Gene/Insert nameSpCas9
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SpeciesS. pyogenes
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Insert Size (bp)4300
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Tag
/ Fusion Protein
- FLAG (N terminal on insert)
Gene/Insert 2
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Gene/Insert nameSp sgRNA scaffold
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Insert Size (bp)75
Gene/Insert 3
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Gene/Insert nameEFS
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Alt nameshort EF1alpha Promoter
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SpeciesH. sapiens (human)
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Insert Size (bp)250
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Entrez GeneEEF1A1 (a.k.a. CCS-3, CCS3, EE1A1, EEF-1, EEF1A, EF-Tu, EF1A, EF1A1, EF1alpha1, GRAF-1EF, LENG7, PTI1, eEF1A-1)
Gene/Insert 4
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Gene/Insert nameP2A-tRFP
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Alt nametagRFP
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SpeciesSynthetic
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Insert Size (bp)750
Resource Information
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Supplemental Documents
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Addgene Notes
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Articles Citing this Plasmid
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
Use BsmBI sites for sgRNA cloning. Note that this plasmid does NOT contain the 1.9kb stuffer from the pLKO.005 vector backbone.
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:
pL-CRISPR.EFS.tRFP was a gift from Benjamin Ebert (Addgene plasmid # 57819) -
For your References section:
Generation of mouse models of myeloid malignancy with combinatorial genetic lesions using CRISPR-Cas9 genome editing. Heckl D, Kowalczyk MS, Yudovich D, Belizaire R, Puram RV, McConkey ME, Thielke A, Aster JC, Regev A, Ebert BL. Nat Biotechnol. 2014 Jun 22. doi: 10.1038/nbt.2951. 10.1038/nbt.2951 PubMed 24952903