AV13_pCAG.Cas9.gRNA.NT
(Plasmid #199260)

• Purpose: Expression construct encoding SpCas9 nuclease and a control non-targeting guide RNA
• Depositing Lab: Manuel Goncalves
• Depositing Organization: Leiden University Medical Center
  Located in Netherlands
• Publication: Wang et al Nucleic Acids Res. 2021 Jan 25;49(2):1173-1198. doi: 10.1093/nar/gkaa1236.

Backbone

• Vector backbone: pBR322
• Backbone size w/o insert (bp): 3196
• Total vector size (bp): 9450
• Vector type: Mammalian Expression, CRISPR

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: SpCas9 nuclease
• Species: Streptococcus pyogenes
• Insert Size (bp): 4137
• Promoter: CAG promoter
• Tag / Fusion Protein:
  • SV40 NLS (C terminal on insert)

Cloning Information for Gene/Insert 1

• Cloning method: Unknown
• 5′ sequencing primer: M13 rev
• 3′ sequencing primer: M13 fwd

Gene/Insert 2

• Gene/Insert name: Non-targeting (NT) control gRNA
• Species: Synthetic
• Insert Size (bp): 20
• Promoter: RNA polymerase III promoter for human U6 snRNA

Cloning Information for Gene/Insert 2

• Cloning method: Restriction Enzyme
• 5′ cloning site: BveI (destroyed during cloning)
• 3′ cloning site: BveI (destroyed during cloning)
• 5′ sequencing primer: M13 rev
• 3′ sequencing primer: M13 fwd

Resource Information

• A portion of this plasmid was derived from a plasmid made by: The SpCas9 insert, derived from Addgene plasmid #41815, is described in: Mali et al. Science 2013 339:823-826.

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

The SpCas9 insert was and placed in a CAG-based expression construct together with an irrelevant non-targeting (NT) gRNA.

DNA (Catalog # 199260-D50)

Purpose

Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.

Delivery

• Amount: 50 μg
• Concentration: 1 μg/μL
• Pricing: $495 USD
• Storage: 4 ℃ (short-term) or -20 ℃ (long-term)

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Quality Control

Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.

Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.

DNA (Catalog # 199260-D100)

Purpose

Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.

Delivery

• Amount: 100 μg
• Concentration: 1 μg/μL
• Pricing: $585 USD
• Storage: 4 ℃ (short-term) or -20 ℃ (long-term)

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Quality Control

Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.

Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.