pII5
(Plasmid #79347)

• Purpose: (Empty Backbone) EcoExpress cloning vector with tac promoter, N term GST tag, and TEV site. For expressing individual protein in E.coli or assembly into position 5 of co-expression vector
• Depositing Lab: Junbiao Dai
• Publication: Qin et al ACS Synth Biol. 2016 Jul 7.

Backbone

• Vector backbone: pET-28b
• Vector type: Bacterial Expression
• Tags / Fusion Proteins:
  • TEV cleavage site (N terminal on backbone)
  • GST (N terminal on backbone)

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): ccdB Survival
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: None
• Mutation: Removed native BsaI site in ccdB gene
• Promoter: tac
• Tag / Fusion Protein:
  • GST (N terminal on backbone)

Cloning Information

• Cloning method: Unknown
• 5′ sequencing primer: M13pUC-rev
• 3′ sequencing primer: T7-term

Terms and Licenses

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

Depositor Comments

Cloning system is based on Golden Gate Assembly, using BsaI for individual vectors and BsmBI for assembly vectors. Assembly is compatible with the BioBrick standard. Oligonucleotide blocking cloning method (OBCM) is used to improve efficiency of multi-insert assembly. See methods in associated article for more details.