pPV-TetO-SpCas9-GR-iC-ERiH (CRONUS-Hygro)
(Plasmid #100597)

• Purpose: piggyBac vector expressing Dual-regulated Cas9 (Hygro resistance)
• Depositing Lab: Akitsu Hotta
• Publication: Ishida et al Sci Rep. 2018 Jan 10;8(1):310. doi: 10.1038/s41598-017-18568-4.

Backbone

• Vector backbone: pPV piggyBac vector
• Backbone manufacturer: Hotta Lab at CiRA, Kyoto University
• Total vector size (bp): 14209
• Vector type: Mammalian Expression ; piggyBac vector
• Selectable markers: Hygromycin

Growth in Bacteria

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

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

• Cloning method: Gateway Cloning
• 5′ sequencing primer: N.A.
• 3′ sequencing primer: N.A.

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.

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.

Resource Information

• Supplemental Documents:
  • pPV-TetO-SphcCas9-hGR-iC-ETiH_addgene.gb

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • piggyBac Limited Use Label License
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry

Depositor Comments

The piggyBac transposase can be obtained directly from Transposagen