pQC hKGA.S286A V5-His IRES G418
(Plasmid #110391)

• Purpose: γ-Retroviral transfer vector for expressing glutminase isoform KGA (S286A), C-terminal V5-6xHis tags, IRES-driven Geneticin selection.
• Depositing Lab: Sandra Martha Gomes Dias
• Publication: HuR (ELAVL1) silencing sensitizes triple-negative breast cancer to glutaminase-targeted therapy (unpublished)

Backbone

• Vector backbone: pQC MCS IRES G418
• Backbone manufacturer: #110344
• Backbone size w/o insert (bp): 7423
• Total vector size (bp): 9561
• Vector type: Mammalian Expression, Retroviral
• 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: KGA glutaminase
• Alt name: GLS (KGA Isoform)
• Species: H. sapiens (human)
• Insert Size (bp): 2007
• Mutation: S286A
• GenBank ID: 2744 NM_014905
• Entrez Gene: GLS (a.k.a. AAD20, CASGID, DEE71, EIEE71, GAC, GAM, GDPAG, GLS1, KGA)
• Promoter: CMV
• Tags / Fusion Proteins:
  • V5 (C terminal on insert)
  • 6xHis (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: AgeI (not destroyed)
• 3′ cloning site: EcoRI (not destroyed)
• 5′ sequencing primer: pQC Seq Fow 5'-ACGCCATCCACGCTGTTTTGACCT-3'
• 3′ sequencing primer: pQC Seq Rev 5'-AAGCGGCTTCGGCCAGTAACGTTA-3'

Resource Information

• Supplemental Documents:
  • pD019-pQC hKGA.S286A V5-His IRES G418.gb

Terms and Licenses

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

Depositor Comments

pQC-based γ-Retroviral transfer vector for KGA.S286A-V5 expression. IRES-driven Puromycin selection.

Weinberg Lab recommends using pUMVC (Addgene #8449) and VSV-G (#8454) for packaging.