pEEK1
(Plasmid #83488)

• Purpose: (Empty Backbone) Expression plasmid for conjugational transfer into cyanobacteria
• Depositing Lab: Pia Lindberg
• Publication: Kukil et al Microb Cell Fact. 2022 Jan 10;21(1):8. doi: 10.1186/s12934-021-01735-8.

Backbone

• Vector backbone: pPMQAK1
• Vector type: Bacterial Expression, Synthetic Biology
• Promoter: Ptrc2O
• Tag / Fusion Protein:
  • Strep-tag (C 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

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ sequencing primer: VF2
• 3′ sequencing primer: VR

Resource Information

• Supplemental Documents:
  • Word document with annotated sequence

Terms and Licenses

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

Depositor Comments

Expression plasmid to overexpress genes in cyanobacteria. Insertion of gene into empty plasmid backbone is based on BglBrick cloning., BglII and BamHI forms a scar inside the ORF that encodes for a serine-glycine linker. Plasmid is inserted into a cyanobacteria by conjugation using the helper cells pRL443. pEEK has a broad host replication origin that has been tested to work in E.coli, Synechocystis 6803, Nostoc 7120 and Nostoc punctiforme.