pBabe-puro-miR-9 sponge
(Plasmid #25040)

• Depositing Lab: Bob Weinberg
• Publication: Ma et al Nat Cell Biol. 2010 Mar . 12(3):247-56.

Backbone

• Vector backbone: pBabe-puro
• Backbone manufacturer: Bob Weinberg, Addgene plasmid 1764
• Backbone size w/o insert (bp): 5169
• Vector type: Mammalian Expression, Retroviral
• Selectable markers: Puromycin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: miR-9 sponge
• Insert Size (bp): 186
• Tag / Fusion Protein:
  • d2EGFP (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (unknown if destroyed)
• 3′ cloning site: SalI (unknown if destroyed)
• 5′ sequencing primer: pBABE 5'
• 3′ sequencing primer: pBABE 3'

Resource Information

• Articles Citing this Plasmid:
  • 5 References

Terms and Licenses

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

Depositor Comments

8 tandem binding sites to human miR-9. Repeated sequence is TCATACAGCTATATACCAAAGA.

Annealed oligonucleotides for tandem miR-9-binding sites were ligated into the pcDNA5-CMV-d2eGFP vector (Invitrogen) digested with XhoI and ApaI. The gfp mRNA, along with the miR-9 sponge sequence in the 3′ UTR was then subcloned into the pBabe-puro vector digested with BamHI and SalI.

Please note that the total insert size between BamHI and SalI is approximately 1327bp, as it includes d2EGFP, the miR-9 binding sites and a BGH polyA terminator.