pDN-Sth1Cas9
(Plasmid #221386)

• Purpose: Cas9 under control of AHT inducible TetR on integrating plasmid (KanR) using L5 integrase and attP for site-specific integration into attB
• Depositing Lab: Deborah Hung
• Depositing Organization: Broad Institute
  Located in the United States of America
• Publication: Neo et al J Bacteriol. 2024 Mar 21;206(3):e0033523. doi: 10.1128/jb.00335-23. Epub 2024 Feb 6.

Backbone

• Vector backbone: pJR962, colE1/phageL5 att/in origins
• Total vector size (bp): 8881
• Modifications to backbone: pJR962 (Addgene plasmid #115162; http://n2t.net/addgene:115162; RRID: Addgene_115162; gift from Jeremy Rock) was used as the template for the construction of the Cas9-containing plasmid (23). Site-directed mutagenesis was performed with the NEBuilder HiFi DNA Assembly Cloning Kit (New England Biolabs) to reintroduce A9D and A599H on the dCas9 allele. These two mutations restore the nuclease activity of the Cas9 allele to facilitate the formation of DSBs. The resulting plasmid pDN-Sth1Cas9 contains genes for the AHT-inducible Sth1Cas9, L5 integrase for integration into the attB site, tetracycline repressor TetR, and resistance selection marker KanR .
• Vector type: CRISPR

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: High copy in E. coli, single copy (integrated) in Mycobacterium
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Sth1 Cas9
• Species: Streptococcus thermophilus
• Insert Size (bp): 3366
• GenBank ID: WP_011680957.1

Terms and Licenses

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

DNA (Catalog # 221386-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 # 221386-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.