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pFGC-pcoCas9
(Plasmid #52256)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 52256 Standard format: Plasmid sent in bacteria as agar stab 1 $65

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pFGC-RCS
  • Modifications to backbone
    expanded multiple cloning sites
  • Vector type
    CRISPR ; Plant expression
  • Selectable markers
    Basta

Growth in Bacteria

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

Gene/Insert

  • Gene/Insert name
    35SPPDK:pcoCas9:NOS cassette
  • Alt name
    Cas9
  • Alt name
    plant codon optimized Cas9
  • Species
    Synthetic
  • Promoter constitutive 35SPPDK promoter

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site StuI (not destroyed)
  • 3′ cloning site StuI (not destroyed)
  • 5′ sequencing primer Sequencing primer for multiple cloning sites (from somewhere close to the EcoRI site): 5’ AATAAAAACTGACTCGGA 3’
  • 3′ sequencing primer Sequencing primer for 35SPPDK:pcoCas9:NOS (from somewhere close to the 35SPPDK promoter): 5’ AGTTGGGTAACGCCAGGG 3’
  • (Common Sequencing Primers)

Resource Information

Depositor Comments

All labeled restriction sites (RS) in red on the map of the binary plasmid pFGC-pcoCas9 is single cut except StuI, which was used to insert the 35SPPDK:pcoCas9:NOS expression cassette. Among those RSs, EcoRI, XhoI, AscI, PacI, AsiSI, SbfI, and XmaI/SmaI can be used to insert single or multiple guide RNA expression cassettes (AscI, PacI, and SbfI are strongly recommended).
The pUC119-gRNA (Addgene plasmid #52255) construct is used as the PCR template to assemble a new guide RNA expression cassette flanked by desired restriction site(s) according to Fig. S5 in Li et al. Nat. Biotechnol. 31: 688-691.

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:

    pFGC-pcoCas9 was a gift from Jen Sheen (Addgene plasmid # 52256 ; http://n2t.net/addgene:52256 ; RRID:Addgene_52256)