pD-St.VA.SN.Mu
(Plasmid #256358)

• Purpose: Tetracistronic transcription unit for SNARE proteins containing 4 Shine-Dalgarno upstream of Syntaxin1A, VAMP2, SNAP25 and Munc18 cDNAs, respectively. Induced by IPTG.
• Depositing Lab: Thierry Galli
• Depositing Organization: UMR INSERM U1266, Institute of Psychiatry and Neuroscience of Paris
  Located in France
• Publication: Ferreras et al J Biol Chem. 2023 Mar;299(3):102974. doi: 10.1016/j.jbc.2023.102974. Epub 2023 Feb 3.

Backbone

• Vector backbone: pRSFDuet-1
• Backbone manufacturer: Novagen
• Vector type: Bacterial Expression

Growth in Bacteria

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

Gene/Insert 1

• Gene/Insert name: Syntaxin 1A
• Species: R. norvegicus (rat)
• Entrez Gene: Stx1a
• Promoter: T7
• Tag / Fusion Protein:
  • 6xHis (N terminal on backbone)

Cloning Information for Gene/Insert 1

• Cloning method: Gibson Cloning
• 5′ sequencing primer: n/a

Gene/Insert 2

• Gene/Insert name: VAMP2
• Species: R. norvegicus (rat)
• Entrez Gene: Vamp2 (a.k.a. RATVAMPB, RATVAMPIR, SYB, Syb2)
• Tag / Fusion Protein:
  • Strep-tagII (N terminal on insert)

Cloning Information for Gene/Insert 2

• Cloning method: Gibson Cloning
• 5′ sequencing primer: n/a

Gene/Insert 3

• Gene/Insert name: SNAP25
• Species: R. norvegicus (rat)
• Entrez Gene: Snap25 (a.k.a. SNAP-25B, SNAP-25a)
• Tag / Fusion Protein:
  • CBP (Calmodulin-binding peptide) (N terminal on insert)

Cloning Information for Gene/Insert 3

• Cloning method: Gibson Cloning
• 5′ sequencing primer: n/a

Gene/Insert 4

• Gene/Insert name: Munc18
• Species: R. norvegicus (rat)
• Entrez Gene: Stxbp2 (a.k.a. Munc18b)
• Tag / Fusion Protein:
  • FLAG (C terminal on insert)

Cloning Information for Gene/Insert 4

• Cloning method: Gibson Cloning
• 5′ sequencing primer: n/a

Terms and Licenses

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