pCK301
(Plasmid
#87767)
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PurposeE. coli rhaBAD promoter upstream of sfGFP, sfGFP can be replaced with any gene of interest to express using L-rhamnose, three terminators, ampR, pBR322 origin, lacI
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 87767 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepJ404
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Backbone manufacturerDNA2.0
- Backbone size w/o insert (bp) 3822
- Total vector size (bp) 4747
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Modifications to backboneT5 promoter replaced with PrhaBAD-sfGFP
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Vector typeBacterial Expression, Synthetic Biology
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 namePrhaBAD-sfGFP
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Alt namerhamnose promoter
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Alt namesuperfolder GFP
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SpeciesE. coli
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Insert Size (bp)925
- Promoter PrhaBAD rhamnose-inducible promoter from E. coli
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Tag
/ Fusion Protein
- 6xHis Tag (C terminal on insert)
Cloning Information
- Cloning method Gibson Cloning
- 5′ sequencing primer TGTAAAACGACGGCCAGT
- 3′ sequencing primer gctcagtcgaaagactgggcctttcgc (Common Sequencing Primers)
Resource Information
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Supplemental Documents
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A portion of this plasmid was derived from a plasmid made bypJT118-sfGFP (Programming Microbes Using Pulse Width Modulation of Optical Signals Davidson, Eric A.; Basu, Amar S.; Bayer, Travis S. Journal of Molecular Biology (2013), 425 (22), 4161-4166 CODEN: JMOBAK; ISSN:0022-2836.)
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Article Citing this Plasmid
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:
pCK301 was a gift from John Heap (Addgene plasmid # 87767 ; http://n2t.net/addgene:87767 ; RRID:Addgene_87767) -
For your References section:
Synthetic Chemical Inducers and Genetic Decoupling Enable Orthogonal Control of the rhaBAD Promoter. Kelly CL, Liu Z, Yoshihara A, Jenkinson SF, Wormald MR, Otero J, Estevez A, Kato A, Marqvorsen MH, Fleet GW, Estevez RJ, Izumori K, Heap JT. ACS Synth Biol. 2016 Oct 21;5(10):1136-1145. Epub 2016 Jun 21. 10.1021/acssynbio.6b00030 PubMed 27247275