pEP4 E02S CK2M EN2L
(Plasmid #20924)

• Purpose: used in the derivation of human iPS cells using non-integrating episomal vectors; expresses Oct4 and Sox2; Klf4 and Myc; Nanog and Lin28
• 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): 11600
• 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 Lin28; Klf4 and Myc
• Species: H. sapiens (human)
• Insert Size (bp): 4900
• GenBank ID: NM_002701 NM_002467 NM_004235 NM_024674 NM_024865 NM_003106
• Entrez Gene: MYC (a.k.a. MRTL, MYCC, bHLHe39, c-Myc)
• Entrez Gene: SOX2 (a.k.a. ANOP3, MCOPS3)
• Entrez Gene: KLF4 (a.k.a. EZF, GKLF)
• Entrez Gene: NANOG
• Entrez Gene: POU5F1 (a.k.a. OCT3, OCT4, OTF-3, OTF3, OTF4, Oct-3, Oct-4, Oct3/4)
• Entrez Gene: LIN28A (a.k.a. CSDD1, LIN-28, LIN28, ZCCHC1, lin-28A)

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:
  • 15 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 Lin28 are coexpressed under a separate (but identical) EF promoter. Klf4 and Myc are coexpressed under a strong CMV 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.

Note that there are some point mutations in the Klf4 gene, but they do not effect the function of this plasmid.