attB Neo* attP
(Plasmid #138412)

• Purpose: attB and attP-flanked pPgk-pEM7-Neo*-bpA positive selection cassette
• Depositing Lab: Mario Capecchi
• Publication: Additional gene targeting vector construction plasmids (unpublished)

Backbone

• Vector backbone: pBluescript
• Backbone manufacturer: Stratagene
• Vector type: Mammalian Expression, Bacterial Expression, Mouse Targeting ; att/PhiC31
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): DH10B
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Neo*
• Species: Synthetic

Resource Information

• Supplemental Documents:
  • attB Neo attP.gb
• A portion of this plasmid was derived from a plasmid made by: The parental plasmid pFNF is derived from plasmid PL451 from Neal Copeland.

Terms and Licenses

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

Depositor Comments

The FRT sites in plasmid pFNF #22687 were replaced by attB and attP sites.

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)