pJ607-CMV-GFP-2A-Cas9
(Plasmid #154830)

• Purpose: GFP-2A-Cas9 (CHO codon-optimized)
• Depositing Lab: Lise Marie Grav
• Depositing Organization: Danmarks Tekniske Universitet
  Located in Denmark
• Publication: Sergeeva et al ACS Synth Biol. 2020 Aug 24. doi: 10.1021/acssynbio.0c00322.

Backbone

• Vector backbone: pJ607
• Backbone manufacturer: DNA2.0
• Backbone size w/o insert (bp): 2541
• Modifications to backbone: Amplified for USER cloning
• Vector type: Mammalian Expression, CRISPR
• Selectable markers: Hygromycin

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: GFP-2A-Cas9
• Insert Size (bp): 4941
• Mutation: Codon-optimized (Cricetulus griseus)
• Promoter: CMV

Cloning Information for Gene/Insert 1

• Cloning method: Unknown
• 5′ sequencing primer: unknown

Gene/Insert 2

• Gene/Insert name: HygR
• Insert Size (bp): 1026
• Promoter: SV40 promoter

Cloning Information for Gene/Insert 2

• Cloning method: Unknown
• 5′ sequencing primer: unknown

Terms and Licenses

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

Depositor Comments

This plasmid contains a codon-optimized Cas9 fused to eGFP for genome editing of Chinese hamster ovary (CHO) cells.

The plasmid was initially described in:
Grav LM et al. 2015 "One-step generation of triple knockout CHO cell lines using CRISPR/Cas9 and fluorescent enrichment" DOI: 10.1002/biot.201500027

A protocol describing the use of the plasmid for CRISPR/Cas9-mediated knockout in CHO cells:
Grav LM et al. 2017 "Application of CRISPR/Cas9 Genome Editing to Improve Recombinant Protein Production in CHO Cells" DOI: 10.1007/978-1-4939-6972-2_7

A protocol describing the use of the plasmid for CRISPR/Cas9-mediated targeted integration in CHO cells:
Sergeeva D et al. 2019 "CRISPR/Cas9 as a Genome Editing Tool for Targeted Gene Integration in CHO Cells". DOI: 10.1007/978-1-4939-9170-9_13

The plasmid was assembled by USER cloning.

DNA (Catalog # 154830-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 # 154830-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.