AAVS1-iSAM
(Plasmid #211495)

• Purpose: AAVS1 targeting vector containing an all-in-one DOX-inducible system for CRISPRa
• Depositing Labs: Antonella Fidanza, Lesley Forrester
• Depositing Organization: University of Edinburgh
  Located in the United Kingdom of Great Britain and Northern Ireland
• Publication: Petazzi et al Elife. 2024 Dec 23;13:RP94884. doi: 10.7554/eLife.94884.
• Citations: 1 Article

Backbone

• Vector backbone: pz Donor
• Total vector size (bp): 15822
• Vector type: Mammalian Expression, CRISPR, Synthetic Biology
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): Stbl3
• Copy number: Unknown

Gene/Insert 1

• Gene/Insert name: UniSAM insert
• Alt name: SV40NLS- dCas9-VP64-MS2-SV40NLS-p65-HSF1-T2A-mCherry
• Species: Synthetic
• Insert Size (bp): 6669
• Promoter: TREGV promoter
• Tag / Fusion Protein:
  • T2A-mCherry Tag

Cloning Information for Gene/Insert 1

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

Gene/Insert 2

• Gene/Insert name: TET-ON
• Alt name: EF1a-TET-ON
• Species: Synthetic
• Insert Size (bp): 747
• Promoter: EF1a

Cloning Information for Gene/Insert 2

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

Gene/Insert 3

• Gene/Insert name: Neo Resistance
• Alt name: SplicingAcceptor-T2A-NeoR
• Species: Synthetic
• Insert Size (bp): 858
• Promoter: Splicing acceptor

Cloning Information for Gene/Insert 3

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

Gene/Insert 4

• Gene/Insert name: Insulator
• Alt name: chicken b-globin insulator
• Species: Synthetic
• Insert Size (bp): 574
• Promoter: NA

Cloning Information for Gene/Insert 4

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

Terms and Licenses

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

Depositor Comments

Please note that the Addgene verified sequence identified differences in the TRE3GV promoter region; however, the plasmid still functions as expected and inducible expression has been tested variety of cell lines (HepG2, iPSCs, hESC, HeLa, HEK).

DNA (Catalog # 211495-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
• 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 # 211495-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
• 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.