pDisplay sHRPa-KDEL
(Plasmid #73149)

• Purpose: Large fragment of split HRP targeted for expression in the lumen of the endoplasmic reticulum (ER). CMV promoter.
• Depositing Lab: Alice Ting
• Publication: Martell et al Nat Biotechnol. 2016 May 30. doi: 10.1038/nbt.3563.

Backbone

• Vector backbone: pDisplay
• Backbone size w/o insert (bp): 5208
• Total vector size (bp): 5979
• Vector type: Mammalian Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: sHRPa-KDEL
• Species: Synthetic
• Insert Size (bp): 771
• Promoter: CMV
• Tag / Fusion Protein:
  • FLAG tag (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BglII (not destroyed)
• 3′ cloning site: AscI (not destroyed)
• 5′ sequencing primer: T7

Terms and Licenses

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

Depositor Comments

sHRPa is the large split HRP fragment. It consists of amino acids 1-213 of horseradish peroxidase (HRP) with the following 4 mutations: T21I, P78S, R93G, N175S.

The insert contains the following features:
EcoRI-Ig k-chain ss-BglII-FLAG tag-sHRPa-3aa linker-KDEL-Stop-AscI

Ig k-chain signal sequence: METDTLLLWVLLLWVPGSTGD
3 aa linker: GSG
FLAG tag: DYKDDDDK