pEGFP-Sec22b-P33-shR
(Plasmid #208359)

• Purpose: Mammalian expression of fluorescent N-terminally tagged shRNA resistant Sec22b-P33 mutant
• Depositing Lab: Paula Nunes-Hasler
• Publication: Criado Santos et al Commun Biol. 2023 Oct 4;6(1):1008. doi: 10.1038/s42003-023-05382-0.

Backbone

• Vector backbone: pEGFP-C1
• Backbone manufacturer: Clonetech
• Backbone size w/o insert (bp): 3930
• Total vector size (bp): 5430
• 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: Sec22b
• Alt name: ERS24
• Alt name: ERS-24
• Species: R. norvegicus (rat)
• Insert Size (bp): 1404
• Mutation: four silent mutations (shRNA resistance), 33 aa proline tract
• GenBank ID: NM_001025686
• Entrez Gene: Sec22b (a.k.a. Sec22l1)
• Promoter: CMV
• Tag / Fusion Protein:
  • EGFP (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: PvuI (not destroyed)
• 3′ cloning site: PvuI (not destroyed)
• 5′ sequencing primer: GTCCTGCTGGAGTTCGTG
• 3′ sequencing primer: CACACCTCCCCCTGAACC

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Thierry Galli, INSERM Paris

Terms and Licenses

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

Depositor Comments

Original plasmid was generated in the laboratory of Dr. Thierry Galli, INSERM Paris (Ref: Petkovic, M., Jemaiel, A., Daste, F. et al. Nat Cell Biol 16, 434–444 (2014). https://doi.org/10.1038/ncb2937) It was cloned into a PvuI site in the pEGFP-Sec22b construct. The plasmid here was generated by site directed mutagenesis using the pEGFP-Sec22b-P33 construct as a template.