lentiGuide-Hygro-dTomato
(Plasmid #99376)

• Purpose: Expresses S. pyogenes CRISPR chimeric RNA element with customizable sgRNA from U6 promoter and hygromycin resistance marker with 2A dTomato from EF-1a promoter. 3rd generation lentiviral backbone.
• Depositing Lab: Kristen Brennand
• Publication: Ho et al Stem Cell Reports. 2017 Aug 8;9(2):615-628. doi: 10.1016/j.stemcr.2017.06.012. Epub 2017 Jul 27.

Backbone

• Vector backbone: lentiGuide ( plasmid #52963)
• Backbone size w/o insert (bp): 9465
• Total vector size (bp): 11394
• Vector type: Mammalian Expression, Lentiviral, CRISPR
• Selectable markers: Hygromycin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): NEB Stable
• Copy number: High Copy

Gene/Insert 1

• Gene/Insert name: S. pyogenes sgRNA cassette
• Species: Synthetic
• Insert Size (bp): 100
• Promoter: hU6

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: BsmBⅠ (not destroyed)
• 3′ cloning site: BsmBⅠ (not destroyed)
• 5′ sequencing primer: hU6-F
• 3′ sequencing primer: hGata4-rev (5'-ATTGTGGATGAATACTGCC-3')

Gene/Insert 2

• Gene/Insert name: Hygro-P2A-dTomato
• Insert Size (bp): 1829
• Promoter: EF-1a

Cloning Information for Gene/Insert 2

• Cloning method: Gibson Cloning
• 5′ sequencing primer: EF-1a-F
• 3′ sequencing primer: WPRE-R

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Hygro: Addgene plasmid #61426 (Feng Zhang lab)
• Articles Citing this Plasmid:
  • 8 References

Terms and Licenses

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

Depositor Comments

Hygro sequence is from Addgene plasmid #61426 (generously shared by Feng Zhang Lab). "Genome-scale transcriptional activation by an engineered CRISPR-Cas9 complex.", Konermann et al.. 2014