pINSPECT:NEAT1-total_SigP:NLuc
(Plasmid #197267)

• Purpose: Donor plasmid for CRISPR/Cas9-mediated insertion of INSPECT into human NEAT1 locus (both isoforms). Encodes for intronic IRES-driven SigP:NLuc. Plasmid contains FRT-site flanked PuroR-TK cassette.
• Depositing Lab: Gil Westmeyer
• Publication: Truong et al Nat Cell Biol. 2022 Nov;24(11):1666-1676. doi: 10.1038/s41556-022-00998-6. Epub 2022 Nov 7.

Backbone

• Vector backbone: pINSPECT_SigP:NLuc
• Backbone size w/o insert (bp): 9113
• Total vector size (bp): 13409
• Vector type: Mammalian Expression, CRISPR, Luciferase, Synthetic Biology, Unspecified
• Selectable markers: Puromycin ; HSV-Thymidine kinase for counter selection using Ganciclovir after removal of selection cassette.

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: NEAT1-total homology arms
• Species: H. sapiens (human)
• Insert Size (bp): 4296

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: T7
• 3′ sequencing primer: T3

Resource Information

• Supplemental Documents:
  • pINSPECT_NEAT1-total_SigP-Nluc.gb

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • NanoLuc Luciferase Limited Use Label License
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Two SNPs present in NEAT1 hom arms will not affect plasmid function.