pQC V5 mKO2 HuR.DelHNS IRES Puro
(Plasmid #110389)

• Purpose: γ-Retroviral transfer vector for expressing HuR (delHNS), V5 and mKO2 tags, IRES-driven Puromycin 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• Publication: HuR (ELAVL1) silencing sensitizes triple-negative breast cancer to glutaminase-targeted therapy (unpublished)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 V5 mKO2 MCS IRES Puro
• Backbone manufacturer: #110341
• Backbone size w/o insert (bp): 7757
• Total vector size (bp): 8603
• Vector type: Mammalian Expression, Retroviral
• Selectable markers: Puromycin

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: ELAV like RNA binding protein 1
• Alt name: ELAVL1
• Alt name: HuR
• Species: H. sapiens (human)
• Insert Size (bp): 981
• Mutation: HNS (HuR nuclear-cytoplasmic shuttling sequence) deleted, 205-237, replaced by lisine-leucine (KL), HindIII site, N121Y
• GenBank ID: 1994 NM_001419
• Entrez Gene: ELAVL1 (a.k.a. ELAV1, HUR, Hua, MelG)
• Promoter: CMV
• Tags / Fusion Proteins:
  • V5 (N terminal on backbone)
  • mKO2 (N terminal on backbone)

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:
  • pD012-pQC V5 mKO2 HuR.DelHNS IRES Puro.gb

Terms and Licenses

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

Depositor Comments

Insert created by joining regions before (Addgene #110395) and after (Addgene #110395) HuR HNS by HindIII restriction site

pQC-based γ-Retroviral transfer vector for V5-HuR.DelHNS expression. IRES-driven Puromycin selection.

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