MIP 227 CoMiP 4in1 wo shRNA p53
(Plasmid #63727)

• Purpose: To express Oct4, Klf4, Sox2, and c-Myc. A single plasmid reprogramming system using a mini-intronic plasmid (MIP).
• Depositing Labs: Mark Kay, Joseph Wu
• Publication: Diecke et al Sci Rep. 2015 Jan 28;5:8081. doi: 10.1038/srep08081.

Backbone

• Vector backbone: codon-optimized minicircle (CoMiC) construct
• Backbone size w/o insert (bp): 7608
• Modifications to backbone: codon-optimized expression cassette of the four canonical reprogramming factors OCT4, KLF4, SOX2, and c-MYC (OKSM)
• Vector type: Mammalian Expression
• Selectable markers: RNA-OUT

Growth in Bacteria

• Bacterial Resistance(s): RNA-OUT ( 6% sucrose)
• Growth Temperature: 30°C
• Growth Strain(s): RNA-OUT selection competent cells, such as NTC4862 DH5α competent cells (Nature Technology)
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Mini-intronic-plasmid intron
• Alt name: pUC-RNA-OUT
• Alt name: MIP intron
• Species: H. sapiens (human)
• Insert Size (bp): 1355
• Mutation: codon-optimised
• Entrez Gene: POU5F1 (a.k.a. OCT3, OCT4, OTF-3, OTF3, OTF4, Oct-3, Oct-4, Oct3/4)
• Entrez Gene: KLF4 (a.k.a. EZF, GKLF)
• Entrez Gene: SOX2 (a.k.a. ANOP3, MCOPS3)
• Entrez Gene: MYC (a.k.a. MRTL, MYCC, bHLHe39, c-Myc)
• Promoter: SFFV
• Tag / Fusion Protein:
  • IRES-dtomato (C terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: AgeI (not destroyed)
• 3′ cloning site: AgeI (not destroyed)
• 5′ sequencing primer: unknown

Resource Information

• Supplemental Documents:
  • annotated sequence MIP227
• A portion of this plasmid was derived from a plasmid made by: The pUC-RNA-OUT sequence used as MIP intron was cloned from NTC vector 8682.

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry

Depositor Comments

In addition to the publication listed above, more details can be found in Lu, et al. Mol Ther. 2013, PMID 23459514.