pFNF
(Plasmid #22687)

• Depositing Lab: Robert Benezra
• Publication: Nam et al Cell Stem Cell. 2009 Nov 6. 5(5):515-26.

Backbone

• Vector backbone: PL451
• Backbone size w/o insert (bp): 5000
• Vector type: Mammalian Expression, Bacterial Expression, Mouse Targeting
• Selectable markers: Neomycin (select with G418)

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: Neo*
• Insert Size (bp): 700
• Mutation: FRT-pPGK-pEM7-Neo*-bpA-FRT

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ sequencing primer: n/a

Resource Information

• A portion of this plasmid was derived from a plasmid made by: N. Copeland
• Articles Citing this Plasmid:
  • 10 References

Terms and Licenses

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

Depositor Comments

The Neo* coding sequence has an E to D mutation at residue 185 corresponding to the original residue 182 (see Yenofsky et al., 1990, https://doi.org/10.1073/pnas.87.9.3435) Its reduced activity could mean that the cell would have to have higher level of the mutant protein expressed in order to achieve drug resistance at the same G418 dose. This could be relevant to mouse embryonic stem cell gene targeting efficiency (also see Hanson and Sedivy, 1995, https://doi.org/10.1128/mcb.15.1.45)