pcDNA3.1-3xFLAG-TEV-UBE2O
(Plasmid #105718)

• Purpose: Expresses WT full length human N- terminally tagged UBE2O
• Depositing Lab: Ramanujan Hegde
• Publication: Yanagitani et al Science. 2017 Aug 4;357(6350):472-475. doi: 10.1126/science.aan0178.

Backbone

• Vector backbone: pcDNA3.1
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 5000
• Total vector size (bp): 8900
• Modifications to backbone: 3xFLAG-TEV was inserted into the vector to serve as a N-terminal tag
• Vector type: Mammalian Expression
• Selectable markers: Zeocin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: UBE2O
• Alt name: ubiquitin conjugating enzyme E2 O
• Species: H. sapiens (human)
• Insert Size (bp): 3876
• GenBank ID: NM_022066.3
• Entrez Gene: UBE2O (a.k.a. E2-230K)
• Promoter: CMV (from vector)
• Tag / Fusion Protein:
  • 3xFLAG-TEV (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoRI (unknown if destroyed)
• 3′ cloning site: XbaI (unknown if destroyed)
• 5′ sequencing primer: GGTCTATATAAGCAGAGC
• 3′ sequencing primer: GCCTGCTATTGTCTTCCC

Resource Information

• A portion of this plasmid was derived from a plasmid made by: UBE2O CDS was originally amplified from pF1KSDA1734 plasmid obtained from Kazusa repository
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

Addgene NGS results found S50F and G1207S within the UBE2O translation compared to NP_071349.3; however the depositing lab confirmed that both of these mutations are known variations and not thought to be detrimental to protein function