Inducible Caspex expression
(Plasmid #97421)

• Purpose: For stable clone generation of cells expressing the dCas9-APEX2 fusion protein for Genomic locus proteomics
• Depositing Labs: Steven Carr, Samuel Myers
• Publication: Myers et al Nat Methods. 2018 May 7. pii: 10.1038/s41592-018-0007-1. doi: 10.1038/s41592-018-0007-1.

Backbone

• Vector backbone: PB-TetON
• Backbone manufacturer: George Church lab
• Backbone size w/o insert (bp): 7501
• Vector type: Mammalian Expression, CRISPR
• 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: Caspex
• Alt name: CATG
• Alt name: Genomic Locus Proteomics
• Species: S. pyogenes, Soybean
• Mutation: dCas9: D10A and H840A, APEX2: K14D, W41F, E112K, A134P
• Promoter: TRE3G
• Tags / Fusion Proteins:
  • 3xFLAG (N terminal on insert)
  • V5 (C terminal on insert)

Cloning Information for Gene/Insert 1

• Cloning method: Ligation Independent Cloning
• 5′ sequencing primer: ggcctccacggccactagt
• 3′ sequencing primer: EF1a-rev

Gene/Insert 2

• Gene/Insert name: T2A-Gfp
• Species: Jellyfish

Resource Information

• Articles Citing this Plasmid:
  • 5 References

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • piggyBac Limited Use Label License
  • Tet Systems Limited Use Label License
• Industry Terms:
  • Not Available to Industry

Depositor Comments

There are ITRs for piggybac transposase insertion but no transposase is in the plasmid itself.

bioRxiv preprint: https://doi.org/10.1101/159517 Posted July 4, 2017.