pJK383
(Plasmid
#72474)
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PurposeProduces Thermoplasma acidophilum citrate synthase with Asp317>Gly mutant (TpCS-D317G)
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 72474 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepRec7-NdeI
- Total vector size (bp) 3764
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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 Temperature30°C
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Growth Strain(s)DH5alpha
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Copy numberUnknown
Gene/Insert
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Gene/Insert namecitrate synthase
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Alt namegltA
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SpeciesThermoplasma acidophilum DSM 1728
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Mutationchanges tryptophan-17 to phenylalanine, tryptophan-115 to phenylalanine, tryptophan-245 to phenylalanine, and aspartate-317 to glycine; silent mutations in leucine-10 and alanine-240.
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GenBank IDTA_RS00855 P21553
- Promoter recA
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site NdeI (destroyed during cloning)
- 3′ cloning site AseI (destroyed during cloning)
- 5′ sequencing primer lambda-PL promoter
- 3′ sequencing primer n/a (Common Sequencing Primers)
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
Protein production is induced using nalidixic acid as described. (Kurz, L. C., Drysdale, G., Riley, M., Tomar, M. A., Chen, J., Russell, R. J. M., and Danson, M. J. ( 2000) Kinetics and mechanism of the citrate synthase from the thermophilic archaeon Thermoplasma acidophilum Biochemistry 39, 2283– 2296 DOI: 10.1021/bi991982r)
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:
pJK383 was a gift from T. J. Kappock (Addgene plasmid # 72474 ; http://n2t.net/addgene:72474 ; RRID:Addgene_72474)