pCK354
(Plasmid #129698)

• Purpose: For insulated expression in Synechocystis 6803. As pCK306 (rhaBAD rhamnose-inducible promoter, YFP, an E. coli-Synechocystis shuttle vector, chromosomal-integration sites) + terminator ECK120010799
• Depositing Lab: John Heap
• Publication: Kelly et al Microorganisms. 2019 Aug 16;7(8). pii: microorganisms7080263. doi: 10.3390/microorganisms7080263.

Backbone

• Vector backbone: pACYC177
• Backbone size w/o insert (bp): 3941
• Total vector size (bp): 6059
• Modifications to backbone: Addition of two 1000bp sequences with homology to the Synechocystis sp. PCC6803 genome, allowing chromosomal integration. Introduction of the rhaBAD promoter from E. coli (allowing inducible expression with L-rhamnose). Introduction of a synthetic RBS and yfp as a reporter. Introduction of rhaS gene from E. coli, the transcriptional activator required for rhaBAD function in Synechocystis.
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Low Copy

Gene/Insert 1

• Gene/Insert name: PrhaBAD
• Alt name: promoter found 5' end of rhaBAD operon in E. coli
• Species: E. coli
• Insert Size (bp): 122

Gene/Insert 2

• Gene/Insert name: yfp
• Alt name: enhanced yellow fluorescent protein from A. victoria (BBa_E0030)
• Species: A. victoria
• Insert Size (bp): 720

Gene/Insert 3

• Gene/Insert name: rhaS
• Alt name: transcriptional activator of the rhaBAD promoter in E. coli with native E. coli RBS
• Species: E. coli
• Insert Size (bp): 837

Gene/Insert 4

• Gene/Insert name: Terminator ECK120010799
• Species: E. coli
• Insert Size (bp): 60

Resource Information

• Supplemental Documents:
  • pCK354.gb

Terms and Licenses

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