pET21-10XHis-GST-HRV-dHer1cys
(Plasmid #73218)

• Purpose: Bacterial expression vector for Z1907 affibody protein (A) with N-terminal His10, GST and HRV3C cleavage site (cysteine tag)
• Depositing Lab: Marcel Bruchez
• Publication: Wang et al Bioconjug Chem. 2015 Jan 21;26(1):137-44. doi: 10.1021/bc500525b. Epub 2014 Dec 19.

Backbone

• Vector backbone: pET21a
• Backbone manufacturer: Novagen
• Backbone size w/o insert (bp): 5387
• Total vector size (bp): 6500
• Modifications to backbone: none
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: For expression use a strain that contains T7 RNA polymerase and trxB/gor double mutant such as Rosetta-gami(DE3)
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: 10XHis-GST-HRV-dHer1cys
• Alt name: dHer1
• Alt name: A
• Species: Synthetic; Schistosoma japonicum
• Insert Size (bp): 1113
• Promoter: T7
• Tags / Fusion Proteins:
  • 10XHis-GST-HRV (N terminal on insert)
  • The dimer of the Her1 Affibody is fused with 4 amino acids between and there is a C terminal cysteine

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NdeI and HindIII for Affibody-Affibody (not destroyed)
• 3′ cloning site: XhoI (not destroyed)
• 5′ sequencing primer: T7 and GSTSeqF
• 3′ sequencing primer: T7term

Terms and Licenses

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

Depositor Comments

Please visit the Molecular Biosensor and Imaging Center website for current protocols and information about dyes to be used in combination with the proteins expressed from this plasmid.

http://pathways.mbic.cmu.edu/