pBABE.puro/TO/Flag/tGFP.HuR.S221D
(Plasmid #110351)

• Purpose: Doxycycline inducible mammalian retroviral expression of HuR (S221D) with FLAG and turboGFP tags
• Depositing Lab: Sandra Martha Gomes Dias
• Publication: Mammalian retroviral expression plasmids based on pBABE family (unpublished)

Backbone

• Vector backbone: pBABE-puro
• Backbone manufacturer: Plasmid #1764
• Backbone size w/o insert (bp): 5177
• Total vector size (bp): 7010
• Modifications to backbone: Sequence obtained from pcDNA5/FRT/TO/Flag-tGFP-HuR.S221D (Addgene #110377) using BglII-XhoI enzymes, containing 1xTO/Flag and tGFP
• 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: S221D, P237S
• GenBank ID: 1994 NM_001419
• Entrez Gene: ELAVL1 (a.k.a. ELAV1, HUR, Hua, MelG)
• Promoter: gag
• Tags / Fusion Proteins:
  • FLAG (N terminal on insert)
  • tGFP (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (destroyed during cloning)
• 3′ cloning site: SalI (destroyed during cloning)
• 5′ sequencing primer: pBABE 5' 5'-CTTTATCCAGCCCTCAC-3'

Resource Information

• Supplemental Documents:
  • pD169-pBABE.puro_TO_Flag_tGFP.HuR.S221D.gb

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Evrogen Limited Use Label License for FPs
• Industry Terms:
  • Not Available to Industry

Depositor Comments

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