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
  • Price
    $65 (USD)
Available to Academic and Nonprofits Only

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
  • Growth Temperature
    37°C
  • Growth Strain(s)
    Top10F'
  • Growth instructions
    Top10 "Extremely" low copy number
  • Copy number
    Low Copy

Sequence Information

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

Cloning Information

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

Resource Information

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.

How to cite this plasmid

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.

  • For your Materials & Methods section:

    pEP4 E02S CK2M EN2L was a gift from James Thomson (Addgene plasmid # 20924)
  • For your References section:

    Human Induced Pluripotent Stem Cells Free of Vector and Transgene Sequences. Yu J, Hu K, Smuga-Otto K, Tian S, Stewart R, Slukvin II, Thomson JA. Science. 2009 Mar 26. 10.1126/science.1172482 PubMed 19325077