AAV-pCbh-SpRY[C-term]-gRNAentry (CA20)
(Plasmid #197511)

• Purpose: Cbh promoter expression plasmid for C-terminal intein-split AAV construct with C-term of SpRY and BsmBI entry cassette to clone SpCas9 gRNA spacer
• Depositing Lab: Benjamin Kleinstiver
• Publication: Alves et al Nat Biomed Eng. 2023 Dec 6. doi: 10.1038/s41551-023-01132-z.

Backbone

• Vector backbone: Cbh_v5 AAV-ABE C-terminal (Addgene Plasmid #137178)
• Vector type: AAV, CRISPR

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): NEB Stable
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: AAV-[ITR]-pCbh-BPNLS-NpuC-SpRY[C-term]-BPNLS-gRNA[BsmBI]-pU6-[ITR]
• Alt name: CA20
• Alt name: C-term AAV-SpRY-gRNAentry
• Species: Synthetic
• Mutation: C-terminal mutations of SpRY(L1111R/D1135L/S1136W/G1218K/E1219Q/N1317R/A1322R/R1333P/R1335Q/T1337R)
• Promoter: Cbh
• Tags / Fusion Proteins:
  • BPNLS-NpuC(intein) (N terminal on insert)
  • BPNLS (C terminal on insert)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: oBK7321-GGGCTGTAATTAGCTGAGCAAGAGG
• 3′ sequencing primer: oBK219-GGGAGTGGCACCTTCCAGGGTC

Resource Information

• Supplemental Documents:
  • AAV-pCbh-SpRY(C-term)-gRNAentry (CA20).gb

Terms and Licenses

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

Depositor Comments

Please visit https://www.biorxiv.org/content/10.1101/2023.01.20.524978v1 for bioRxiv preprint.