U6_sgRNA_CAG_ABE7.10_St1Cas9_LMD9
(Plasmid #136660)

• Purpose: Expresses ABE St1Cas9 LMD9 in mammalian cells along with its U6-driven sgRNA
• Depositing Lab: Yannick Doyon
• Publication: Agudelo et al Genome Res. 2020 Jan;30(1):107-117. doi: 10.1101/gr.255414.119. Epub 2020 Jan 3.

Backbone

• Vector backbone: MSP1594_2x_NLS (Plasmid #110625)
• Total vector size (bp): 8715
• Vector type: Mammalian Expression, CRISPR, Synthetic Biology

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: St1Cas9 LMD-9
• Species: H. sapiens (human), Synthetic
• Promoter: CAG
• Tags / Fusion Proteins:
  • SV40 NLS (N terminal on insert)
  • SV40 NLS (C terminal on insert)

Cloning Information for Gene/Insert 1

• Cloning method: Gibson Cloning
• 5′ sequencing primer: ctcagaagaaggcccagagc

Gene/Insert 2

• Gene/Insert name: sgRNA for St1Cas9
• Species: H. sapiens (human), Synthetic
• Promoter: U6

Cloning Information for Gene/Insert 2

• Cloning method: Gibson Cloning
• 5′ sequencing primer: taaaattatgttttaaaatggac

Gene/Insert 3

• Gene/Insert name: ABE7.10
• Alt name: TadA-linker(32aa)-TadA*(7.10)-linker(32aa)-nCas9-NLS
• Species: H. sapiens (human), Synthetic; Escherichia coli
• Promoter: CAG
• Tag / Fusion Protein:
  • hSt1Cas9 (C terminal on insert)

Cloning Information for Gene/Insert 3

• Cloning method: Gibson Cloning
• 5′ sequencing primer: ggttattgtgctgtctcatcat

Resource Information

• Supplemental Documents:
  • U6_sgRNA_CAG_NLS_ABE_St1Cas9_NLS.gb

Terms and Licenses

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

Depositor Comments

A U6-driven sgRNA expression cassette was cloned upstream of the CAG promoter in MSP1594_2x_NLS (Plasmid # 110625). This plasmid is a derivative of MSP1594 (Plasmid # 110625). Please visit https://www.biorxiv.org/content/early/2018/05/14/321208 for BioRxiv preprint