pDONR223_HyPer7RgDAAO
(Plasmid #217746)

• Purpose: Donor vector for gateway cloning of HyPer7 DAAO, a fusion protein used to measure the uptake of D-amino acids across the plasma membrane.
• Depositing Lab: Seth Parker
• Publication: Johal et al Cell Reports, Volume 43, Issue 12, 115069

Backbone

• Vector backbone: pDONR223
• Total vector size (bp): 5370
• Modifications to backbone: Inserted HyPer7 Rg DAAO fusion between the att sites.
• Vector type: Mammalian Expression

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: HyPer7 D amino acid oxidase
• Alt name: HyPer7 DAAO
• Species: Neisseria meningitidis, Rhodotorula garcilis
• Insert Size (bp): 2638
• Mutation: Fused DAAO to the C-terminus of HyPer7 using a Gly-Gly-Ser-Gly link while also removing the ATG start codon for DAAO
• Tag / Fusion Protein:
  • Nuclear export signal (C terminal on insert)

Cloning Information

• Cloning method: Gateway Cloning
• 5′ sequencing primer: TGTAAAACGACGGCCAGT
• 3′ sequencing primer: CAGGAAACAGCTATGAC

Resource Information

• Supplemental Documents:
  • pDONR223_HyPer7RgDAAO.gbk
• A portion of this plasmid was derived from a plasmid made by: Valeriy V Pak and Vsevolod Belousov

Terms and Licenses

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