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PurposeEgg cell-specific promoter-controlled expression of 3×FLAG-NLS-zCas9-NLS. Contains gRNA scaffold for insertion of target sequence (U6-26 promoter), Hyg resistance
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 71287 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepCambia
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Vector typePlant Expression, CRISPR ; Plant binary vector
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Selectable markersHygromycin
Growth in Bacteria
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Bacterial Resistance(s)Kanamycin, 50 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 1
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Gene/Insert namegRNA scaffold
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SpeciesSynthetic
- Promoter U6-26p Arabidopsis U6 gene promoter
Cloning Information for Gene/Insert 1
- Cloning method Restriction Enzyme
- 5′ cloning site BsaI (not destroyed)
- 3′ cloning site BsaI (not destroyed)
- 5′ sequencing primer M13-fwd (Common Sequencing Primers)
Gene/Insert 2
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Gene/Insert namezCas9
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Alt name3xFLAG-NLS-zCas9-NLS
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SpeciesS. pyogenes
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MutationZea mays codon-optimized Cas9
- Promoter EC1.2 enhancer fused to EC1.1 promoter
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Tags
/ Fusion Proteins
- 3x FLAG (N terminal on insert)
- NLS (N terminal on insert)
- NLS (C terminal on insert)
Cloning Information for Gene/Insert 2
- Cloning method Unknown
- 5′ sequencing primer n/a
- 3′ sequencing primer M13-rev (Common Sequencing Primers)
Resource Information
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Supplemental Documents
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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
Please see the cloning protocol attachments for detailed descriptions on how to insert sgRNA sequences into this plasmid.
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:
pHEE401E was a gift from Qi-Jun Chen (Addgene plasmid # 71287 ; http://n2t.net/addgene:71287 ; RRID:Addgene_71287) -
For your References section:
Egg cell-specific promoter-controlled CRISPR/Cas9 efficiently generates homozygous mutants for multiple target genes in Arabidopsis in a single generation. Wang ZP, Xing HL, Dong L, Zhang HY, Han CY, Wang XC, Chen QJ. Genome Biol. 2015 Jul 21;16:144. doi: 10.1186/s13059-015-0715-0. 10.1186/s13059-015-0715-0 [pii] PubMed 26193878