Venus-7aa-Cb5-pEGFP-C1
(Plasmid #166764)

• Purpose: To target Venus to the Endoplasmic Reticulum. Cb5 tail anchor inserts into the membrane and Venus faces the cytosol. This serves as a collisional control in FLIM-FRET experiments.
• Depositing Lab: David Andrews
• Publication: Andrew lab Venus and Cerulean fusions (unpublished)

Backbone

• Vector backbone: pEGFP-C1
• 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: Cb5 tail anchor
• Alt name: Venus-Cb5, V-Cb5
• Species: R. norvegicus (rat)
• Entrez Gene: Cyb5a (a.k.a. Cyb5)
• Promoter: CMV
• Tag / Fusion Protein:
  • Venus (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: Unknown (unknown if destroyed)
• 3′ cloning site: Unknown (unknown if destroyed)
• 5′ sequencing primer: TGACGCAAATGGGCGGTAGG
• 3′ sequencing primer: TCGCCCTTTGACGTTGGAGTCCAC

Resource Information

• Supplemental Documents:
  • Venus-7aa-Cb5-pEGFP-C1.gb
• A portion of this plasmid was derived from a plasmid made by: See gene entry for "cytochrome b5 [Rattus norvegicus]", See NCBI Reference Sequence: NP_071581.1. Here we have used only the C-terminal tail anchor sequence.

Terms and Licenses

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

Depositor Comments

"KITTVESNSSWWTNWVIPAISALVVALMYRLYMAED" amino acid targeting sequence, which we refer to as "Cb5", for endoplasmic reticulum targeting. "7aa" indicates the 7 amino acid flexible linker region between Venus and the Cb5 targeting signal.