FH95pUp95bGW (B5)
(Plasmid #74014)

• Purpose: lentiviral expression of shRNA resistant Psd95-beta-EGFP fusion and Psd95 shRNA
• Depositing Labs: Robert Malenka, Oliver Schluter, Weifeng Xu
• Publication: Schluter et al Neuron. 2006 Jul 6;51(1):99-111.

Backbone

• Vector backbone: FUGW
• Backbone manufacturer: David Baltimore (Addgene plasmid # 14883)
• Backbone size w/o insert (bp): 9941
• Vector type: Mammalian Expression, Lentiviral, RNAi

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: Psd95 shRNA
• Alt name: shRNA-resistant Psd95-EGFP fusion co-expressed
• gRNA/shRNA sequence: GGA CAT CCA GGC ACA CAA G
• Species: R. norvegicus (rat)
• Entrez Gene: Dlg4 (a.k.a. Dlgh4, PSD95, Sap90)
• Promoter: H1

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoRI (not destroyed)
• 3′ cloning site: BstBI (destroyed during cloning)
• 5′ sequencing primer: H1 tcgctatgtgttctgggaaa

Terms and Licenses

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

Depositor Comments

Map: FHUG+W-HinDIII-SalI-XbaI-(p95b)-KpnI-SacII-PspOMI-SmaI-BamHI-(GFP)-BglII-BsrBI-EcoRV-HinDIII-ClaI
Also associated with Liu, et al. J Neurophysiol. 2014 Feb;111(3):648-58. doi: 10.1152/jn.00262.2013 PMID 24225540.

Psd95 shRNA expressed under control of H1 promoter. Psd95 coding sequence modified to GGATATTCAAGCGCATAAG make it resistant to the shRNA. Psd95-EGFP fusion protein expressed under control of UbC promoter.

The publication by Schluter et al linked above describes the creation of the beta isoform of Psd95 in detail