pEP4 E02S EN2K
(Plasmid #20925)

• Purpose: used in the derivation of human iPS cells using non-integrating episomal vectors; expresses Oct4 and Sox2; Nanog and Klf4
• Depositing Lab: James Thomson
• Publication: Yu et al Science. 2009 Mar 26.

Backbone

• Vector backbone: pCEP4
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 10500
• Vector type: Episomal/EBNA

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): Top10F'
• Growth instructions: Top10 "Extremely" low copy number
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: Oct4 and Sox2; Nanog and Klf4
• Species: H. sapiens (human)
• Insert Size (bp): 5700
• GenBank ID: NM_002701 NM_004235 NM_024865 NM_003106
• Entrez Gene: SOX2 (a.k.a. ANOP3, MCOPS3)
• Entrez Gene: POU5F1 (a.k.a. OCT3, OCT4, OTF-3, OTF3, OTF4, Oct-3, Oct-4, Oct3/4)
• Entrez Gene: NANOG
• Entrez Gene: KLF4 (a.k.a. EZF, GKLF)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: multiple (unknown if destroyed)
• 3′ cloning site: multiple (unknown if destroyed)
• 5′ sequencing primer: na

Resource Information

• Addgene Notes:
  • Addgene diagnostic digest
• Articles Citing this Plasmid:
  • 21 References

Terms and Licenses

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

Depositor Comments

Oct4 and Sox2 are coexpressed under a strong EF promoter. Nanog and Klf4 are coexpressed under a separate (but identical) EF promoter. This plasmid contains EBNA-1 and OriP for limited maintenance in mammalian cells.

These plasmids were used at concentrations of >1.5 mg/ml. When preparing these plasmids for transfection, 3 low concentration maxipreps are done and the elution is combined in about 700 ul.