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PurposeExpression of His6-MBP-tagged Cas9-NLS-EGFP protein in E. coli
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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 | 78312 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepET His6 MBP N10 TEV LIC cloning vector (2C-T)
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Backbone manufacturerScott Gradia
- Backbone size w/o insert (bp) 5938
- Total vector size (bp) 10927
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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 numberLow Copy
Gene/Insert
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Gene/Insert nameSpCas9
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Alt nameCas9
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Alt nameCsn1
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SpeciesStreptococcus pyogenes
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Insert Size (bp)4989
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Mutationhuman codon-optimized gene sequence
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GenBank IDNP_269215
- Promoter T7
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Tags
/ Fusion Proteins
- His6-MBP-TEV (N terminal on backbone)
- HA-2xNLS-EGFP-NLS (C terminal on insert)
Cloning Information
- Cloning method Ligation Independent Cloning
- 5′ sequencing primer MalE FW
- 3′ sequencing primer T7 terminator (Common Sequencing Primers)
Resource Information
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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.
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:
pMJ922 was a gift from Martin Jinek (Addgene plasmid # 78312 ; http://n2t.net/addgene:78312 ; RRID:Addgene_78312) -
For your References section:
Maximizing mutagenesis with solubilized CRISPR-Cas9 ribonucleoprotein complexes. Burger A, Lindsay H, Felker A, Hess C, Anders C, Chiavacci E, Zaugg J, Weber LM, Catena R, Jinek M, Robinson MD, Mosimann C. Development. 2016 Apr 29. pii: dev.134809. 10.1242/dev.134809 PubMed 27130213