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pHEE401E
(Plasmid #71287)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 71287 Standard format: Plasmid sent in bacteria as agar stab 1 $75

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pCambia
  • Vector type
    Plant Expression, CRISPR ; Plant binary vector
  • Selectable markers
    Hygromycin

Growth in Bacteria

  • Bacterial Resistance(s)
    Kanamycin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    High Copy

Gene/Insert 1

  • Gene/Insert name
    gRNA scaffold
  • Species
    Synthetic
  • 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

  • Gene/Insert name
    zCas9
  • Alt name
    3xFLAG-NLS-zCas9-NLS
  • Species
    S. pyogenes
  • Mutation
    Zea mays codon-optimized Cas9
  • Promoter EC1.2 enhancer fused to EC1.1 promoter
  • 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

Resource Information

Depositor Comments

Please see the cloning protocol attachments for detailed descriptions on how to insert sgRNA sequences into this plasmid.

How to cite this plasmid ( Back to top)

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.

  • 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