MSCV FLIPi P2G_Thy1.1 Dbl (p53,PTEN)
(Plasmid #19746)

• Depositing Lab: Richard Hynes
• Publication: Stern et al Proc Natl Acad Sci U S A. 2008 Sep 16. 105(37):13895-900.

Backbone

• Vector backbone: MSCV FLIPi
• Backbone size w/o insert (bp): 7994
• Vector type: Mammalian Expression, Retroviral, RNAi, Cre/Lox
• Selectable markers: Puromycin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): Stbl3
• Growth instructions: grow in stbl3 cells at 30oC
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: oligo targeting p53 and PTEN
• gRNA/shRNA sequence: none given
• Species: M. musculus (mouse)
• Insert Size (bp): 778
• Entrez Gene: Pten (a.k.a. 2310035O07Rik, A130070J02Rik, B430203M17Rik, MMAC1, PTENbeta, TEP1)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NotI (not destroyed)
• 3′ cloning site: PmeI (not destroyed)
• 5′ sequencing primer: GFP miR primer (GCTGGAGTTCGTGACCGCC)

Resource Information

• Supplemental Documents:
  • FLIP vector protocols
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Constitutively expresses puromycin-resistance and GFP. Recombines to express surface Thy1.1 and miR30-based RNAi. The miRNA is contained within an artificial intron, and splicing of the intron can boost transgene expression (1.5 to 2-fold). The expression of surface Thy1.1 facilitates antibody-mediated cell sorting.

See FLIP vector protocols (PDF from this page) for more information.
There are minor discrepancies between the depositor's sequence and Addgene's sequencing results. The mismatches are either coding joints or a sequencing difference between the actual and the published miR30 vector sequence.