pGAGmEos2-N1
(Plasmid #46372)

• Depositing Lab: Suliana Manley
• Publication: Gunzenhauser et al Nano Lett. 2012 Sep 12;12(9):4705-10. doi: 10.1021/nl3021076. Epub 2012 Aug 27.

Backbone

• Vector backbone: tdEos-N1
• Backbone manufacturer: Shroff, et al PNAS 2007 PMID: 18077327
• Backbone size w/o insert (bp): 4054
• Modifications to backbone: made by replacing EGFP with tdEos in EGFP-N1 (clonetech).
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Gag
• Alt name: HIV assembly protein
• Species: Human immunodeficiency virus 1
• Mutation: Rev independentHXB2 Gag sequence
• Promoter: CMV
• Tag / Fusion Protein:
  • mEos2 (C terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: XhoI (not destroyed)
• 3′ cloning site: BamHI (not destroyed)
• 5′ sequencing primer: CMV-F
• 3′ sequencing primer: SV40pA-R

Terms and Licenses

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

Depositor Comments

The plasmid encoding for Gag-tdEos was cloned as described [Manley et al, Nature Methods (2008)]. The HIV-1 assembly protein Gag (a Rev independentHXB2 Gag sequence) was excised from pGagEGFP2 (gift from Dr.Marilyn Resh, Memorial Sloan-Kettering Cancer Center, New York, NY, USA) and inserted into tdEos-N1 via XhoI/BamHI sites. Gag-mEos2, replacing the tdEos gene, was constructed by subcloning the BamHI/XbaI fragment from pmEos2-N1 (provided by G.Patterson [McKinney SA Nat. Methods]) into the plasmid encoding for Gag-tdEos after digestion with BamHI/XbaI.