pEP4 E02S EM2K
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||20923||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
- Backbone size w/o insert (bp) 12600
Growth in Bacteria
Growth instructionsTop10 "Extremely" low copy number
Copy numberLow Copy
Gene/Insert nameOct4 and Sox2; Myc and Klf4
SpeciesH. sapiens (human)
Insert Size (bp)2700
GenBank IDNM_002701 NM_004235 NM_002467 NM_003106
Entrez GeneSOX2 (a.k.a. ANOP3, MCOPS3)
Entrez GeneKLF4 (a.k.a. EZF, GKLF)
Entrez GeneMYC (a.k.a. MRTL, MYCC, bHLHe39, c-Myc)
Entrez GenePOU5F1 (a.k.a. OCT3, OCT4, OTF-3, OTF3, OTF4, Oct-3, Oct-4)
- 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)
Oct4 and Sox2 are coexpressed under a strong EF promoter. Myc 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.
Note that this a K278M mutation in Klf4, but it does not affect the reprogramming ability.
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 EM2K was a gift from James Thomson (Addgene plasmid # 20923 ; http://n2t.net/addgene:20923 ; RRID:Addgene_20923)
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