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PurposeTn7 integrating plasmid with S. pasteurianus dCas9 expressed from the Plac promoter for expression in P. aeruginosa
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 105235 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepUC18
- Backbone size w/o insert (bp) 5981
- Total vector size (bp) 9404
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Vector typeBacterial Expression, CRISPR, Synthetic Biology
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Selectable markersGentamicin
Growth in Bacteria
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Bacterial Resistance(s)Ampicillin, 100 μg/mL
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Growth Temperature37°C
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Growth Strain(s)DH5alpha
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert nameS. pasteurianus dCas9
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SpeciesS. pasteurianus
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Insert Size (bp)3423
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MutationD10A, H599A
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Tag
/ Fusion Protein
- HA (N terminal on insert)
Resource Information
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Supplemental Documents
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A portion of this plasmid was derived from a plasmid made byS. 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
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
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For your Materials & Methods section:
pUC18-mini-Tn7T-Plac-dCas9 (Plac) was a gift from Kristala Prather (Addgene plasmid # 105235 ; http://n2t.net/addgene:105235 ; RRID:Addgene_105235) -
For your References section:
A Robust CRISPR Interference Gene Repression System in Pseudomonas. Tan SZ, Reisch CR, Prather KLJ. J Bacteriol. 2018 Mar 12;200(7). pii: JB.00575-17. doi: 10.1128/JB.00575-17. Print 2018 Apr 1. 10.1128/JB.00575-17 PubMed 29311279