pAB1677 REPAIR.t3
(Plasmid #176320)

• Purpose: CMV-HIVNES-GS-dCas13bt3- (GGS)2-huADAR2dd(E488Q) (REPAIR.t3 expression)
• Depositing Lab: Feng Zhang
• Publication: Kannan et al Nat Biotechnol. 2021 Aug 30. pii: 10.1038/s41587-021-01030-2. doi: 10.1038/s41587-021-01030-2.

Backbone

• Vector backbone: pcDNA3.1
• Total vector size (bp): 8919
• Vector type: Mammalian Expression, CRISPR
• Selectable markers: Neomycin (select with G418)

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 1

• Gene/Insert name: human codon optimized Cas13bt3 (catalytically inactivated)
• Species: Planctomycetes bacterium isolate B28_G16 (marine sediment)
• Insert Size (bp): 2322
• Mutation: R84A/H89A/R740A/H744A
• GenBank ID: RKY08123.1
• Promoter: CMV
• Tag / Fusion Protein:
  • HIV NES (N terminal on insert)

Cloning Information for Gene/Insert 1

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

Gene/Insert 2

• Gene/Insert name: huADAR2dd(E488Q)
• Alt name: human ADAR2 deaminase domain
• Species: H. sapiens (human)
• Insert Size (bp): 1158
• Mutation: E488Q, only the deaminase domain (aa 276-702 are expressed)
• Entrez Gene: ADARB1 (a.k.a. ADAR2, DRABA2, DRADA2, NEDHYMS, RED1)

Resource Information

• Supplemental Documents:
  • pab1677-cmv-hivnes-dcas13b-t3-ggs2-huadar2dde488q.gb

Terms and Licenses

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