Venus-30aa-Noxa-4E-pEGFP-C1
(Plasmid #167529)

• Purpose: To increase the length of the linker between Venus and Noxa-4E proteins
• 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• Publication: Andrew lab Venus and Cerulean fusions (unpublished)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: PMA-induced protein 1
• Alt name: Noxa
• Species: H. sapiens (human)
• Mutation: C25E, L29E, F32E, L36E
• Entrez Gene: PMAIP1 (a.k.a. APR, NOXA)
• Promoter: CMV
• Tag / Fusion Protein:
  • Venus (N terminal on insert)

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

Terms and Licenses

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

Depositor Comments

The Noxa sequence aligns with phorbol-12-myristate-13-acetate-induced protien 1 isoform 6 [Homo sapiens] (NCBI Reference Sequence: NM_021127.3)

Venus fused to the N-terminus of human Noxa amino acid sequence: "MPGKKARKNAQPSPARAPAELEVECATQLRRFGDKLNFRQKLLNLISKLFCSGT". Linked by a 30 amino acid (aa) linker. Noxa BH3 region has hydrophobic positions H1-H4 mutated to glutamic acid (E), referred to as "BH3-4E" mutation, BH3 mutations: C25E, L29E, F32E, L36E. When compared with EYFP, the Venus fluorophore contains the following mutations: F46L, F64L, M153T, V163A and S175G. This Venus protein contains an A207K, F224R and L232H mutations to make it monomeric. An additional A164V mutation in Venus occurred in this construct, which does not appear to affect fluorescence of the protein.