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Addgene

pJK048
(Plasmid #75230)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 75230 Standard format: Plasmid sent in bacteria as agar stab 1 $85
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Backbone

  • Vector backbone
    pET23a
  • Backbone size w/o insert (bp) 3666
  • Total vector size (bp) 5191
  • Vector type
    Bacterial Expression

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    BL21(DE3)
  • Copy number
    Unknown

Gene/Insert 1

  • Gene/Insert name
    N5-carboxyaminoimidazole ribonucleotide mutase
  • Alt name
    purE
  • Alt name
    JW0512
  • Species
    Escherichia coli str. K-12 substr. MG1655
  • Mutation
    changes histidine-45 to tryptophan
  • GenBank ID
    AAC73625
  • Promoter T7

Cloning Information for Gene/Insert 1

  • Cloning method Restriction Enzyme
  • 5′ cloning site NdeI (not destroyed)
  • 3′ cloning site n/a (unknown if destroyed)
  • 5′ sequencing primer T7 promoter
  • 3′ sequencing primer n/a
    • (Common Sequencing Primers)

Gene/Insert 2

  • Gene/Insert name
    N5-carboxyaminoimidazole ribonucleotide synthetase
  • Alt name
    purK
  • Alt name
    JW0511
  • Species
    Escherichia coli str. K-12 substr. MG1655
  • Mutation
    Pro64 to Arg; Asp65 to His; Gln206 to Arg
  • GenBank ID
    AAC73624
  • Promoter n/a

Cloning Information for Gene/Insert 2

  • Cloning method Restriction Enzyme
  • 5′ cloning site n/a (unknown if destroyed)
  • 3′ cloning site HindIII (not destroyed)
  • 5′ sequencing primer n/a
  • 3′ sequencing primer T7 terminator
    • (Common Sequencing Primers)

Resource Information

  • A portion of this plasmid was derived from a plasmid made by
    Constructed by T. J. Kappock in the laboratory of JoAnne Stubbe (MIT).

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry
Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

This plasmid encodes a purK gene with arginine-64 and histidine-65 codons, an apparently natural variant in some E. coli accessions, as well as a glutamine-205 to arginine mutant introduced during assembly of the parental plasmid pNC2 (Addgene #72438)..

How to cite this plasmid ( Back to top)

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.

  • For your Materials & Methods section:

    pJK048 was a gift from T. J. Kappock & JoAnne Stubbe (Addgene plasmid # 75230 ; http://n2t.net/addgene:75230 ; RRID:Addgene_75230)

  • For your References section:

    N5-CAIR mutase: role of a CO2 binding site and substrate movement in catalysis. Hoskins AA, Morar M, Kappock TJ, Mathews II, Zaugg JB, Barder TE, Peng P, Okamoto A, Ealick SE, Stubbe J. Biochemistry. 2007 Mar 13;46(10):2842-55. Epub 2007 Feb 14. 10.1021/bi602436g PubMed 17298082
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