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Purpose"All-in-one" CRISPR/Cas9 (wt) plasmid for cloning of custom gRNA with blue/white screening
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 74293 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepCRISPR-CG01
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Backbone manufacturerGenecopedia
- Backbone size w/o insert (bp) 10415
- Total vector size (bp) 10694
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Vector typeMammalian Expression, Bacterial Expression, CRISPR
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Selectable markersNeomycin (select with G418)
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 numberUnknown
Gene/Insert 1
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Gene/Insert nameLacZ-alpha
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SpeciesSynthetic
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Insert Size (bp)279
- Promoter Lac promoter
Cloning Information for Gene/Insert 1
- Cloning method Gateway Cloning
- 5′ sequencing primer SP6 Forward: ATTTAGGTGACACTATAGA
- 3′ sequencing primer none (Common Sequencing Primers)
Gene/Insert 2
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Gene/Insert nameCas9
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Alt nameCsn1
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SpeciesStreptococcus pyogenes
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Insert Size (bp)4104
- Promoter T7 promoter
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Tag
/ Fusion Protein
- HA (C terminal on insert)
Cloning Information for Gene/Insert 2
- Cloning method Unknown
- 5′ sequencing primer T7 promoter (NOT UNIQUE IN PLASMID) (Common Sequencing Primers)
Gene/Insert 3
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Gene/Insert namemCherry
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Speciesunknown
- Promoter SV40
Cloning Information for Gene/Insert 3
- Cloning method Unknown
- 5′ sequencing primer unknown (Common Sequencing Primers)
Resource Information
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Article 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
See Supporting information file 1: "Instructions for one-step gRNA cloning into All-in-one CRISPR/Cas9 LacZ" from associated publication for gRNA cloning instructions using this plasmid.
Please cite associated publication when using this plasmid for cloning.
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:
All_in_one_CRISPR/Cas9_LacZ was a gift from Lynne Postovit (Addgene plasmid # 74293 ; http://n2t.net/addgene:74293 ; RRID:Addgene_74293) -
For your References section:
A Digital PCR-Based Method for Efficient and Highly Specific Screening of Genome Edited Cells. Findlay SD, Vincent KM, Berman JR, Postovit LM. PLoS One. 2016 Apr 18;11(4):e0153901. doi: 10.1371/journal.pone.0153901. eCollection 2016. PONE-D-16-05005 [pii] PubMed 27089539