pEEK2
(Plasmid #83492)

• Purpose: (Empty Backbone) Expression plasmid for conjugational transfer into cyanobacteria
• Depositing Lab: Pia Lindberg
• Publication: Miao et al Metab Eng Commun. 2017 Jul 29;5:45-53. doi: 10.1016/j.meteno.2017.07.003. eCollection 2017 Dec.

Backbone

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

Cloning Information

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

Resource Information

• Supplemental Documents:
  • Word document with annotated sequence
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

Expression plasmid to overexpress genes in cyanobacteria using bicistronic design from Mutalik et al. 2013. 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 with the use of helper cells pRL443. pEEK2 has a broad host replication origin that has been tested to work in E.coli, Synechocystis 6803, Nostoc 7120 and Nostoc punctiforme.