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Addgene

pJK_proNR2_LGK.1
(Plasmid #81161)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 81161 Standard format: Plasmid sent in bacteria as agar stab 1 $85
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Backbone

  • Vector backbone
    pJK
  • Backbone manufacturer
    pET29 derivative
  • Backbone size w/o insert (bp) 3582
  • Total vector size (bp) 4899
  • Modifications to backbone
    Removal of the T7 promoter, T7 operator, lacI gene. Addition of the constitutive promoter proJK1 between BglI and XbaI. LGK is between NdeI and XhoI.
  • Vector type
    Bacterial Expression

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Growth instructions
    Rich media for plasmid production as the constitutive promoter will reduce production under minimal media.
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    Levoglucosan kinase
  • Alt name
    B3VI55_LIPST
  • Species
    Synthetic; Lipomyces starkeyi
  • Insert Size (bp)
    1317
  • Mutation
    LGK.1 (L140I,S142A,A373C)
  • GenBank ID
    ACE79748.1 EU751287.1
  • Promoter proNR2
  • Tag / Fusion Protein
    • His6x (C terminal on backbone)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site NdeI (not destroyed)
  • 3′ cloning site XhoI (not destroyed)
  • 5′ sequencing primer GGTGATTCATTCTGCTAA
  • 3′ sequencing primer T7 Term
    • (Common Sequencing Primers)

Terms and Licenses

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

    pJK_proNR2_LGK.1 was a gift from Timothy Whitehead (Addgene plasmid # 81161 ; http://n2t.net/addgene:81161 ; RRID:Addgene_81161)

  • For your References section:

    Comprehensive sequence-flux mapping of a levoglucosan utilization pathway in E. coli. Klesmith JR, Bacik JP, Michalczyk R, Whitehead TA. ACS Synth Biol. 2015 Sep 14. 10.1021/acssynbio.5b00131 PubMed 26369947
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