pLNGY-PML.I-2K1R-3D
(Plasmid #260897)

• Purpose: Expresses human full length PML I triple mutant with an N-terminal EGFP tag in mammalian cells
• Depositing Lab: Irina Bezsonova
• Publication: PXL: a Nucleic Acid–Binding Module of Promyelocytic Leukemia Protein (unpublished)

Backbone

• Vector backbone: pLKO.1
• Total vector size (bp): 10661
• Modifications to backbone: The BamHI-KpnI fragment encoding puromycin resistance was removed from pLKO-shPML1 and replaced with a PCR fragment encoding G418 resistance flanked by the same restriction sites, to create pLKOneo-shPML1. Lentivirus vector plasmids expressing proteins from the HSV-1 gD (US6) gene promoter were constructed by removing the RNA polymerase III and shRNA sequences from pLKOneo.shPML1 and replacing them with a fragment from −122 to +11 of the HSV-1 gD promoter (Everett, 1983) linked to the EYFP coding region. Plasmids with this backbone were designated with the prefix pLNGY. The open reading frames of PML isoforms I to VI, as defined (Jensen et al., 2001), were inserted downstream of the EYFP sequences to give plasmid pLNGY-PML.I
• Vector type: Mammalian Expression, Lentiviral
• Selectable markers: Neomycin (select with G418)

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: PML.I
• Species: H. sapiens (human)
• Insert Size (bp): 2648
• Mutation: K678D, K725D, R792D
• Promoter: human U6
• Tag / Fusion Protein:
  • EYFP (N terminal on insert)

Cloning Information

• Cloning method: Gene Synthesis
• 5′ sequencing primer: EGFP-C

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Roger D Everett

Terms and Licenses

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