pAAVS1_TREtight-FRB-GEM23-Halo-IRES-FKBP-SNAP-GFPNb_EF1a-rtTA3-T2A-Puro-P2A-Thy1.1
(Plasmid #197065)

• Purpose: Donor plasmid for human AAVS1 locus knock-in, doxycycline-inducible expression of FRB-GEM23-Halo and Adaptor Protein (FKBP-SNAP-vhhGFP4)
• Depositing Lab: Julia Mahamid
• Publication: Fung et al Nat Methods. 2023 Dec;20(12):1900-1908. doi: 10.1038/s41592-023-02053-0. Epub 2023 Nov 6.

Backbone

• Vector backbone: pSH-EFIRES-P
• Vector type: Mammalian Expression, CRISPR
• Selectable markers: Puromycin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Unknown

Gene/Insert 1

• Gene/Insert name: I3-01
• Alt name: GEM23
• Species: Synthetic
• Mutation: Codon-optimised for human cell expression
• GenBank ID:
• Tags / Fusion Proteins:
  • FRB (N terminal on insert)
  • HaloTag (C terminal on insert)

Gene/Insert 2

• Gene/Insert name: Adaptor Protein
• Alt name: FKBP-SNAP-vhhGFP4

Resource Information

• A portion of this plasmid was derived from a plasmid made by: The protein sequence for nanocage I3-01 was derived from Addgene Plasmid #80046, first reported in Hsia, Y. et al. Design of a hyperstable 60-subunit protein icosahedron. Nature 535, 136–139 (2016). Addgene plasmids #129715 and #60510 were used in the creation of this plasmid. Anti-GFP nanobody vhhGFP4 was derived from Addgene #117713.
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • HaloTag Vector Limited Use Label License
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Please visit https://www.biorxiv.org/content/10.1101/2022.12.10.519870v1 for bioRxiv preprint.