pAAVS1_TREtight-GEM2-Halo-FRB-IRES-FKBP-SNAP-GFPNb_EF1a-rtTA3-T2A-Puro-P2A-Thy1.1
(Plasmid #197061)

• Purpose: Donor plasmid for human AAVS1 locus knock-in, doxycycline-inducible expression of GEM2-Halo-FRB and Adaptor Protein (FKBP-SNAP-vhhGFP4)
• Depositing Lab: Julia Mahamid
• Depositing Organization: European Molecular Biology Laboratory (EMBL), Heidelberg
  Located in Germany
• 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: Encapsulin
• Alt name: GEM2
• Species: Synechococcus elongatus
• Mutation: Codon-optimised for human cell expression
• GenBank ID:
• Tags / Fusion Proteins:
  • HaloTag (C terminal on insert)
  • FRB (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: Addgene plasmids #129715 and #60510 were used in the creation of this plasmid. Anti-GFP nanobody vhhGFP4 was derived from Addgene #117713.

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.

DNA (Catalog # 197061-D50)

Purpose

Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.

Delivery

• Amount: 50 μg
• Concentration: 1 μg/μL
• Pricing: $495 USD
• Storage: 4 ℃ (short-term) or -20 ℃ (long-term)

Terms and Licenses

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

Quality Control

Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.

Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.

DNA (Catalog # 197061-D100)

Purpose

Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.

Delivery

• Amount: 100 μg
• Concentration: 1 μg/μL
• Pricing: $585 USD
• Storage: 4 ℃ (short-term) or -20 ℃ (long-term)

Terms and Licenses

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

Quality Control

Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.

Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.