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pPV-TetO-SpCas9-GR-iC-ERiP (CRONUS-Puro)
(Plasmid #100596)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 100596 Standard format: Plasmid sent in bacteria as agar stab 1 $85
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Backbone

  • Vector backbone
    pPV piggyBac vector
  • Backbone manufacturer
    Hotta Lab at CiRA, Kyoto University
  • Total vector size (bp) 14246
  • Vector type
    Mammalian Expression ; piggyBac vector
  • Selectable markers
    Puromycin

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    Unknown

Gene/Insert 1

  • Gene/Insert name
    CRISPR Cas9 fused with human Glucocorticoid Receptor
  • Alt name
    SpCas9-hGR
  • Species
    H. sapiens (human); Streptococcus pyogenes
  • Insert Size (bp)
    4920
  • Mutation
    Codon-optimized for human codon usage
  • Promoter Dox-inducible TetO promoter

Cloning Information for Gene/Insert 1

Gene/Insert 2

  • Gene/Insert name
    mCherry
  • Species
    Discosoma sp.
  • Insert Size (bp)
    711
  • Promoter Dox-inducible TetO promoter

Cloning Information for Gene/Insert 2

  • Cloning method Restriction Enzyme
  • 5′ cloning site ApaI (not destroyed)
  • 3′ cloning site PacI (destroyed during cloning)
  • 5′ sequencing primer N.A.
  • 3′ sequencing primer N.A.
    • (Common Sequencing Primers)

Gene/Insert 3

  • Gene/Insert name
    rtTA-M2
  • Species
    Synthetic; Escherichia coli
  • Insert Size (bp)
    747
  • Promoter Human EEF1A1 promoter

Cloning Information for Gene/Insert 3

  • Cloning method Restriction Enzyme
  • 5′ cloning site PacI (destroyed during cloning)
  • 3′ cloning site EcoRV (destroyed during cloning)
  • 5′ sequencing primer N.A.
  • 3′ sequencing primer N.A.
    • (Common Sequencing Primers)

Resource Information

Terms and Licenses

Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

The piggyBac transposase can be obtained directly from Transposagen

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:

    pPV-TetO-SpCas9-GR-iC-ERiP (CRONUS-Puro) was a gift from Akitsu Hotta (Addgene plasmid # 100596 ; http://n2t.net/addgene:100596 ; RRID:Addgene_100596)

  • For your References section:

    Site-specific randomization of the endogenous genome by a regulatable CRISPR-Cas9 piggyBac system in human cells. Ishida K, Xu H, Sasakawa N, Lung MSY, Kudryashev JA, Gee P, Hotta A. Sci Rep. 2018 Jan 10;8(1):310. doi: 10.1038/s41598-017-18568-4. 10.1038/s41598-017-18568-4 PubMed 29321585
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