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PurposeRNA-based iPSC generation. VEE RNA replicon 3'ORF encoding OCT4, KLF4, SOX2, separated by 2A peptides followed by an IRES then c-myc and a second IRES and PuroR ORF
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 58973 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backboneSP6-VEE-IRES-puro (Addgene plasmid 58971)
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Backbone manufacturerDowdy lab
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Vector typeMammalian Expression ; Venezuelan equine encephalitis (VEE) virus RNA replicon
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Selectable markersPuromycin
Growth in Bacteria
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Bacterial Resistance(s)Ampicillin, 100 μg/mL
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Growth Temperature37°C
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Growth Strain(s)DH5alpha
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Copy numberUnknown
Gene/Insert 1
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Gene/Insert nameOct4
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SpeciesH. sapiens (human)
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Entrez GenePOU5F1 (a.k.a. OCT3, OCT4, OTF-3, OTF3, OTF4, Oct-3, Oct-4, Oct3/4)
Gene/Insert 2
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Gene/Insert nameKlf4
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Alt nameKruppel-like factor 4
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SpeciesH. sapiens (human)
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Entrez GeneKLF4 (a.k.a. EZF, GKLF)
Gene/Insert 3
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Gene/Insert nameSox2
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Alt nameSRY (sex determining region Y)-box 2
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SpeciesH. sapiens (human)
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Entrez GeneSOX2 (a.k.a. ANOP3, MCOPS3)
Gene/Insert 4
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Gene/Insert namecMyc
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SpeciesH. sapiens (human)
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Entrez GeneMYC (a.k.a. MRTL, MYCC, bHLHe39, c-Myc)
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
Depositor Comments
Summary of features:
1-7561: VEE RNA replicases (nsP1, nsP2, nsP3, nsP4 and 26S promoter, nsP1~4 are nonstructural VEE sequences which encode RNA replicases)
7592-8671: human Oct4
8678-8731: T2A peptide sequence
8738-10147: human Klf4
10154-10213: E2A peptide sequence
10223-11176: human Sox2
11195-11805: IRES sequence
11818-13140: human cMyc
13165-13776: IRES sequence
13777-14376: Puromycin resistance gene
14383-14510:VEE 3’UTR and poly A tail
14679-15539: Ampicillin resistance gene
16320-16337: SP6 promoter
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.
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For your Materials & Methods section:
SP6-VEE-OKS-iM was a gift from Steven Dowdy (Addgene plasmid # 58973 ; http://n2t.net/addgene:58973 ; RRID:Addgene_58973) -
For your References section:
Efficient generation of human iPSCs by a synthetic self-replicative RNA. Yoshioka N, Gros E, Li HR, Kumar S, Deacon DC, Maron C, Muotri AR, Chi NC, Fu XD, Yu BD, Dowdy SF. Cell Stem Cell. 2013 Aug 1;13(2):246-54. doi: 10.1016/j.stem.2013.06.001. 10.1016/j.stem.2013.06.001 PubMed 23910086