pLVX-TetOn-puro-ICP4-RR/LL
(Plasmid #250614)

• Purpose: Mammalian lentiviral vector for the inducible (TetOn) expression of codon-optimized HSV-1 ICP4 with a double amino acid substitution in the site-specific DNA binding domain (R456L-R457L)
• Depositing Lab: Liam Holt
• Publication: Herpes simplex virus 1 fluidizes the nucleus enabling condensate formation (unpublished)

Backbone

• Vector backbone: pLVX-TetOn-puro
• Backbone size w/o insert (bp): 9180
• Total vector size (bp): 13079
• Modifications to backbone: Insertion of two overlapping fragments from codon-optimized HSV-1 ICP4 at the MCS
• Vector type: Mammalian Expression, Lentiviral
• Selectable markers: Puromycin

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: ICP4-RR/LL
• Alt name: mDBD
• Alt name: RR/LL
• Alt name: R456L-R457L
• Species: Herpes simplex virus 1 (KOS strain)
• Insert Size (bp): 3885
• Mutation: ICP4 contains two point mutations in the site-specific DNA binding domain: R456L & R457L
• Promoter: TRE3GS promoter

Cloning Information

• Cloning method: Other
• 5′ sequencing primer: cgtaccacttcctaccctcgtaaagaattcgcaggctcttaaggccaccA
• 3′ sequencing primer: tcattggctgtccagcttagctcGCAGGGGAGGTGGTCTGggagggagaggggcatcg

Resource Information

• Supplemental Documents:
  • pLH2658.gb
• A portion of this plasmid was derived from a plasmid made by: Cloned codon-optimized ICP4 from Addgene #250609 (pTRE-ICP4-BFP-Neo)

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Tet Systems Limited Use Label License
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Point mutations were introduced with overlapping overhangs and NEB HiFi DNA Assembly Cloning.
Please visit https://doi.org/10.1101/2025.06.20.660750 for bioRxiv preprint.