pSLPB2B-FKBP12_F36V-CasRx-tagBFP-PGK-Blasticidin
(Plasmid #187952)

• Purpose: FKBP12 (F36V mutant) degron-tagged CasRx fused with tagBFP, expressed under EF1-alpha promoter in a piggyBac plasmid. Expresses Blasticidin resistance under PGK promoter.
• Depositing Lab: Edda Schulz
• Publication: Noviello et al Nat Commun. 2023 Jun 3;14(1):3225. doi: 10.1038/s41467-023-38909-4.

Backbone

• Vector backbone: pSLPB2-FKBP12_F36VSpdCas9-tagBFP-PGK-Blasticidin
• Backbone size w/o insert (bp): 6727
• Total vector size (bp): 9817
• Vector type: Mammalian Expression, CRISPR, Synthetic Biology
• Selectable markers: Blasticidin

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: FKBP12_F36V-CasRx-tagBFP
• Alt name: RfxCas13d
• Alt name: FKBP12 (F36V mutant)
• Species: H. sapiens (human), Synthetic; R. flavifaciens
• Insert Size (bp): 3090
• Tags / Fusion Proteins:
  • GGS linker (C terminal on backbone)
  • GGS linker (N terminal on insert)
  • HA-tag (C terminal on insert)
  • NLS (N terminal on insert)
  • NLS (C terminal on insert)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: GCGGATCTGGAGGTAGTGGT
• 3′ sequencing primer: CAGATCCACCACTACCTCCAG

Resource Information

• A portion of this plasmid was derived from a plasmid made by: The CasRx domain was amplified from pLentiRNACRISPR_007 - TetO-NLS-RfxCas13d-NLS-WPRE-EFS-rtTA3-2A-Blast from Neville Sanjana (Addgene plasmid # 138149 ; http://n2t.net/addgene:138149 ; RRID:Addgene_138149)

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Evrogen Limited Use Label License for FPs
  • piggyBac Limited Use Label License
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Please visit https://www.biorxiv.org/content/10.1101/2022.10.05.511019v1 for bioRxiv preprint.