pUC18-mini-Tn7T-Lac-dCas9 (Ptac)
(Plasmid #105234)

• Purpose: Tn7 integrating plasmid with S. pasteurianus dCas9 expressed from the Ptac promoter for expression in P. putida and P. fluorescens
• Depositing Lab: Kristala Prather
• Publication: Tan et al J Bacteriol. 2018 Mar 12;200(7). pii: JB.00575-17. doi: 10.1128/JB.00575-17. Print 2018 Apr 1.

Backbone

• Vector backbone: pUC18
• Backbone size w/o insert (bp): 5979
• Total vector size (bp): 9402
• Vector type: Bacterial Expression, CRISPR, Synthetic Biology
• Selectable markers: Gentamicin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: S. pasteurianus dCas9
• Species: S. pasteurianus
• Insert Size (bp): 3423
• Mutation: D10A, H599A
• Tag / Fusion Protein:
  • HA (N terminal on insert)

Resource Information

• Supplemental Documents:
  • pUC18-mini-Tn7T-Lac-dCas9 (Ptac) wps.ape
• A portion of this plasmid was derived from a plasmid made by: S. pasteurianus dCas9 was kindly provided by Rock et al. 2017. Programmable transcriptional repression in mycobacteria using an orthogonal CRISPR interference platform. Nat Microbiol 2:16274. pUC18-mini-Tn7T-Lac was derived from Choi et al. 2006. mini-Tn7 insertion in bacteria with single attTn7 sites: example Pseudomonas aeruginosa. Nat Protoc 1:153–161.

Terms and Licenses

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