U6_sgRNA_CAG_APOBEC-1_hSt1Cas9_TH1477_2xUGI_3xHA
(Plasmid #136659)

• Purpose: Expresses St1BE4max-TH1477 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 LMD9:TH1477 chimera
• Species: H. sapiens (human), Synthetic
• Promoter: CAG
• Tags / Fusion Proteins:
  • SV40 NLS (N terminal on insert)
  • SV40 NLS (C terminal on insert)
  • APOBEC-1 (N terminal on insert)
  • 2xUGI (C terminal on insert)

Cloning Information for Gene/Insert 1

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

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: gagggcctatttcccatgattcct

Gene/Insert 3

• Gene/Insert name: APOBEC-1
• Species: H. sapiens (human)
• Entrez Gene: APOBEC1 (a.k.a. APO1, APOBEC-1, BEDP, CDAR1, HEPR)
• Promoter: CAG
• Tag / Fusion Protein:
  • hSt1Cas9 (C terminal on insert)

Cloning Information for Gene/Insert 3

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

Gene/Insert 4

• Gene/Insert name: UGI
• Species: H. sapiens (human), Synthetic
• Promoter: CAG
• Tag / Fusion Protein:
  • hSt1Cas9 (N terminal on insert)

Cloning Information for Gene/Insert 4

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

Gene/Insert 5

• Gene/Insert name: HA tag
• Species: Synthetic
• Tag / Fusion Protein:
  • UGI (N terminal on insert)

Resource Information

• Supplemental Documents:
  • U6_sgRNA_CAG_APOBEC_hSt1Cas9_TH1477_2xUGI_3xHA.gb

Terms and Licenses

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