pDEST-SHmOCR
(Plasmid #206874)

• Purpose: (Empty Backbone) Empty Gateway pDEST vector for bacterial expression with Super His - Monomeric OCR fusion
• Depositing Lab: Murray Junop
• Publication: Szabla et al Nucleic Acids Res. 2024 Jul 22:gkae635. doi: 10.1093/nar/gkae635.

Backbone

• Vector backbone: pDest-527
• Backbone manufacturer: Dominic Esposito (Addgene plasmid # 11518)
• Backbone size (bp): 8695
• Modifications to backbone: Replaced 6xHis tag on pDEST-527 with 14xHis (SuperHis) + monomeric OCR (mOCR) fusion via BglII sites
• Vector type: Bacterial Expression
• Promoter: T7
• Tags / Fusion Proteins:
  • SuperHis (14xHis) + monomeric OCR (mOCR) (N terminal on backbone)
  • HSV + 6xHis (C terminal on backbone)

Growth in Bacteria

• Bacterial Resistance(s): Chloramphenicol and Ampicillin, 25 & 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): ccdB Survival
• Growth instructions: This is an empty Gateway pDEST vector - must propagate in ccdB-resistant strain such as DB3.1
• Copy number: High Copy

Cloning Information

• Cloning method: Gateway Cloning
• 5′ sequencing primer: T7 promoter
• 3′ sequencing primer: T7 terminator

Resource Information

• Supplemental Documents:
  • pMJ5973 - pDEST-SuperHis-mOCR - standard genbank.gb
  • Snapgene plasmid map - refer to "history" tab for plasmid construction details (previously loaded)

Terms and Licenses

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

Depositor Comments

Useful bacterial expression vector for purifying challenging proteins.
SuperHis (14xHis) ensures very high affinity to Ni-IMAC column.
mOCR tag is very negatively charged. This promotes proper folding, prevents aggregation and enables purification via ion-exchange.

Refer to DelProposto et al 2008 (DOI: 10.1016/j.pep.2008.08.011) for information on mOCR tag fusions.