pL-CRISPR.EFS.tRFP
(Plasmid #57819)

• Purpose: Lentiviral CRISPR-Cas9 delivery for SpCas9 and sgRNA. Coexpresses tagRFP via P2A cleavage site. EFS Promoter driven
• Depositing Lab: Benjamin Ebert
• Publication: Heckl et al Nat Biotechnol. 2014 Jun 22. doi: 10.1038/nbt.2951.

Backbone

• Vector backbone: pLKO.005
• Total vector size (bp): 11701
• Vector type: Lentiviral, CRISPR

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): Stbl3
• Copy number: High Copy

Gene/Insert 1

• Gene/Insert name: SpCas9
• Species: S. pyogenes
• Insert Size (bp): 4300
• Tag / Fusion Protein:
  • FLAG (N terminal on insert)

Gene/Insert 2

• Gene/Insert name: Sp sgRNA scaffold
• Insert Size (bp): 75

Gene/Insert 3

• Gene/Insert name: EFS
• Alt name: short EF1alpha Promoter
• Species: H. sapiens (human)
• Insert Size (bp): 250
• Entrez Gene: EEF1A1 (a.k.a. CCS-3, CCS3, EE1A1, EEF-1, EEF1A, EF-Tu, EF1A, EF1A1, EF1alpha1, GRAF-1EF, LENG7, PTI1, eEF1A-1)

Gene/Insert 4

• Gene/Insert name: P2A-tRFP
• Alt name: tagRFP
• Species: Synthetic
• Insert Size (bp): 750

Resource Information

• Supplemental Documents:
  • Target site cloning protocol
• Addgene Notes:
  • Addgene Diagnostic Digest
• Articles Citing this Plasmid:
  • 26 References

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Evrogen Limited Use Label License for FPs
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Use BsmBI sites for sgRNA cloning. Note that this plasmid does NOT contain the 1.9kb stuffer from the pLKO.005 vector backbone.