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This material is available to academics and nonprofits only.

Backbone

• Vector backbone
  lentiCRISPR (pXPR_001), Addgene plasmid #49535
  (Search Vector Database)
• Backbone manufacturer
  Zhang lab
• Vector type
  Mammalian Expression, Lentiviral, CRISPR
• Selectable markers
  Puromycin

Growth in Bacteria

• Bacterial Resistance(s)
  Ampicillin
• Growth Temperature
  37°C
• Growth Strain(s)
  Stbl3
• Growth instructions
  Use SapI digest to check for unwanted recombination of lentiviral plasmid. Only amplify in RecA- bacteria (eg. Stbl3).
• Copy number
  High Copy

Gene/Insert1

• Gene/Insert name
  Cas9
• Alt name
  S. pyogenes CRISPR-Cas9
• Species
  Synthetic
• Insert Size (bp)
  4200
• PromoterEFS

Cloning Informationfor Gene/Insert 1

• Cloning methodRestriction Enzyme
• 5′ cloning siteBamHI(not destroyed)
• 3′ cloning siteNheI(not destroyed)
• 5′ sequencing primerGGTACAGTGCAGGGGAAAGA
• 3′ sequencing primerTGCCCTCCAAATATGTGAACT
(Common Sequencing Primers)

Gene/Insert2

• Gene/Insert name
  Puromycin resistance
• Alt name
  puromycin N-acetyl-transferase
• Alt name
  PAC
• Insert Size (bp)
  600
• PromoterEFS

Cloning Informationfor Gene/Insert 2

• Cloning methodRestriction Enzyme
• 5′ cloning siteNheI(not destroyed)
• 3′ cloning siteMluI(not destroyed)
• 5′ sequencing primerTGCTGCTACTAAGAAAGCTGGTC
(Common Sequencing Primers)

Gene/Insert3

• Gene/Insert name
  EGFP sgRNA 4
• Alt name
  EGFP targeting RNA element #4 (with +85 chimeric RNA)
• Species
  Synthetic
• Insert Size (bp)
  96
• PromoterU6

Cloning Informationfor Gene/Insert 3

• Cloning methodRestriction Enzyme
• 5′ cloning siteBsmBI(destroyed during cloning)
• 3′ cloning siteBsmBI(destroyed during cloning)
• 5′ sequencing primer#140 hU6-F
(Common Sequencing Primers)

Resource Information

• Terms and Licenses
  • UBMTA
• Industry Terms
  • Not Available to Industry
• Articles Citing this Plasmid
  • 2 References

Depositor Comments