Addgene - pEP4 E02S CK2M EN2L Plasmid Data

Plasmid 20924: pEP4 E02S CK2M EN2L

Gene/insert name:
Oct4 and Sox2; Nanog and Lin28; Klf4 and Myc
Insert size:
4900
Species:
H. sapiens (human)
GenBank ID:
NM_002701 NM_002467 NM_004235 NM_024674 NM_024865 NM_003106
Vector backbone:
pCEP4
(Search Vector Database)
Backbone manufacturer:
Invitrogen
Vector type:
Episomal/EBNA
Backbone size w/o insert (bp):
11600
Cloning site 5':
multiple
Site destroyed during cloning:
Unknown
Cloning site 3':
multiple
Site destroyed during cloning:
Unknown
5' sequencing primer:
na List of Sequencing Primers
Bacterial resistance(s)
Ampicillin
Growth strain(s)
Top10F'
Growth temperature (℃):
37
Growth instructions:
Top10 "Extremely" low copy number
High or low copy:
Low Copy
Sequence: View sequences (4)
Map:
View map
Supplemental document:
Notes from Addgene (1)
Principal Investigator:
James Thomson
Terms and Licenses:
MTA

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.

Addgene has sequenced a portion of this plasmid for verification. Click here for the sequencing result.

Article: Human Induced Pluripotent Stem Cells Free of Vector and Transgene Sequences Yu et al (Science. 2009 Mar 26. PubMed)

Please acknowledge the principal investigator and cite this article if you use this plasmid in a publication. Also, please include the text "Addgene plasmid 20924" in your Materials and Methods section.