pEGFP VAMP7 (102-220)
(Plasmid #42314)

• Depositing Lab: Thierry Galli
• Publication: Martinez-Arca et al J Cell Biol. 2000 May 15;149(4):889-900.

Backbone

• Vector backbone: pEGFP-C1
• Backbone manufacturer: Clontech
• Backbone size w/o insert (bp): 4700
• 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: VAMP7 Coiled Coil domain (102-220)
• Alt name: VAMP7
• Alt name: vesicle-associated membrane protein 7
• Alt name: TI-VAMP/Synaptobrevin-like (Sybl 1)
• Species: H. sapiens (human)
• Mutation: Δ1-102
• GenBank ID: NM_005638.5 NP_005629.1
• Entrez Gene: VAMP7 (a.k.a. SYBL1, TI-VAMP, TIVAMP, VAMP-7)
• Promoter: CMV
• Tag / Fusion Protein:
  • EGFP (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: SalI (not destroyed)
• 3′ cloning site: SmaI (destroyed during cloning)
• 5′ sequencing primer: EGFP-C; CMV-F
• 3′ sequencing primer: SV40-pA-R

Terms and Licenses

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

Depositor Comments

Alternate plasmid name: ΔNH2-terminal domain-TIVAMP (ΔNter-TIVAMP, from M102 to the end)

For production of NH2-terminal GFP fusion proteins, the distinct cDNAs were cloned into the pEGFP-C3 vector (CLONTECH Laboratories, Inc.). Full-length TI-VAMP (TIVAMP), NH2-terminal domain-TIVAMP (Nter-TIVAMP, from M to N120), cytosolic domain-TIVAMP (Cyt-TIVAMP, from M to K188), and ΔNH2-terminal domain-TIVAMP (ΔNter-TIVAMP, from M102 to the end), were obtained by PCR using standard procedures and the following sets of oligonucleotides:

5′-ATGGCGATTCTTTTTGCTGTTGTTGCC-3′
and
5′-CTATTTCTTCACACAGCTTGGCCATGT-3′
for TIVAMP;

5′-ATGGCGATTCTTTTTGCTGTTGTTGCC-3′
and
5′-CTTATTCTCAGAGTGATGCTTCAG-CTG-3′
for Nter-TIVAMP;

5′-ATGGCGATTCTTTTTGCTGTTGTTGCC-3′
and
5′-ATCCTACTTGAGGTTCTTCATACACATGGCTC
for Cyt-TIVAMP;

5′-ATGAATAGCGAGTTCTCAAGTGTCTTA-3′
and
5′-CTATTTCTTCACACAGCTTGGCCATGT-3′
for ΔNter-TIVAMP.

TIVAMP, Nter-TIVAMP, and Cyt-TIVAMP were cloned in pEGFP-C3 using the KpnI-XbaI sites, while ΔNter-TIVAMP was cloned in HindIII-XbaI sites.