Skip to main content
Addgene

pNC2
(Plasmid #72438)

Print

Ordering

This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 72438 Standard format: Plasmid sent in bacteria as agar stab 1 $85
Add to Cart

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)
    DH5alpha
  • Growth instructions
    Overnight growth to saturation leads to leaky expression of PurE and PurK.
  • Copy number
    High Copy

Gene/Insert 1

  • Gene/Insert name
    N5-carboxyaminoimidazole ribonucleotide mutase
  • Alt name
    purE
  • Alt name
    JW0512
  • Species
    Escherichia coli str. K-12 substr. MG1655
  • GenBank ID
    AAC73625.1
  • 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
    AAC73624.1
  • 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
    A portion of this plasmid was derived from work done 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

Construction details: http://dx.doi.org/10.1021/bi9827159.

This plasmid encodes arginine-64 and histidine-65, an apparently natural variant in some E. coli accessions, as well as a glutamine-205 to arginine mutant introduced during plasmid assembly.

This plasmid encodes purK with arginine-64: Thoden, J.B., Kappock, T.J., Stubbe, J., Holden, H.M. (1999) "Three-dimensional structure of N5-carboxyaminoimidazole ribonucleotide synthetase: a member of the ATP grasp protein superfamily." Biochemistry 38: 15480-15492. PMID: 10569930 DOI: 10.1021/bi991618s

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:

    pNC2 was a gift from T. J. Kappock & JoAnne Stubbe (Addgene plasmid # 72438 ; http://n2t.net/addgene:72438 ; RRID:Addgene_72438)

  • For your References section:

    Evidence for the direct transfer of the carboxylate of N5-carboxyaminoimidazole ribonucleotide (N5-CAIR) to generate 4-carboxy-5-aminoimidazole ribonucleotide catalyzed by Escherichia coli PurE, an N5-CAIR mutase. Meyer E, Kappock TJ, Osuji C, Stubbe J. Biochemistry. 1999 Mar 9;38(10):3012-8. 10.1021/bi9827159 PubMed 10074353
Related items: