EcPhnD90.L1∆∆AD.L297RL301R.pRSET
(Plasmid
#33376)
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Purposea single-wavelength phosphonate sensor constructed from E. coli PhnD and cpGFP
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Depositing Lab
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Publication
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 33376 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
This material is available to academics and nonprofits only.
Backbone
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Vector backbonepRSET
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Backbone manufacturerInvitrogen
- Backbone size w/o insert (bp) 2900
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Vector typeBacterial Expression
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 namephnD based Phosphonate Biosensor
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SpeciesE. coli
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Insert Size (bp)1668
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MutationcpGFP146 variant from GCaMP2 inserted in place of AA89-90 of phnD, L297R in phnD.
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Entrez GenephnD (a.k.a. B21_03937)
- Promoter T7
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Tag
/ Fusion Protein
- 6xHIS (N terminal on backbone)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site BamHI (not destroyed)
- 3′ cloning site EcoRI (not destroyed)
- 5′ sequencing primer T7 forward (Common Sequencing Primers)
Resource Information
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Supplemental Documents
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.
Depositor Comments
The EcPhnD-cpGFP insertion variants were constructed by overlap PCR using the synthetic EcPhnD sequence and the cpGFP146 variant from GCaMP2. Detailed sequences are provided in attached sheets. The linker regions were mutated by single-stranded template mutagenesis
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:
EcPhnD90.L1∆∆AD.L297RL301R.pRSET was a gift from Loren Looger (Addgene plasmid # 33376 ; http://n2t.net/addgene:33376 ; RRID:Addgene_33376) -
For your References section:
Structure of the Escherichia coli Phosphonate Binding Protein PhnD and Rationally Optimized Phosphonate Biosensors. Alicea I, Marvin JS, Miklos AE, Ellington AD, Looger LL, Schreiter ER. J Mol Biol. 2011 Oct 12. 10.1016/j.jmb.2011.09.047 PubMed 22019591