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Addgene

pETconNK-LGK
(Plasmid #81165)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 81165 Standard format: Plasmid sent in bacteria as agar stab 1 $85
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Backbone

  • Vector backbone
    pETconNK
  • Backbone size w/o insert (bp) 6109
  • Total vector size (bp) 7423
  • Modifications to backbone
    Starting with pETcon the ampicillin resistance gene was swapped with kanamycin with the BbvCI nicking site 3' of the KanR gene. The nicking sequence CCTCAGCAA is on the coding strand.
  • Vector type
    Bacterial Expression, Yeast Expression
  • Selectable markers
    TRP1

Growth in Bacteria

  • Bacterial Resistance(s)
    Kanamycin, 50 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    Levoglucosan kinase wild-type
  • Alt name
    B3VI55_LIPST
  • Species
    Synthetic; Lipomyces starkeyi
  • Insert Size (bp)
    1317
  • GenBank ID
    ACE79748.1 EU751287.1
  • Promoter GAL1
  • Tag / Fusion Protein
    • c-myc (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 GGACAATAGCTCGACGATTGAAGGTAGATACCCATA
  • 3′ sequencing primer TAATACGACTCACTATAGG
    • (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:

    pETconNK-LGK was a gift from Timothy Whitehead (Addgene plasmid # 81165 ; http://n2t.net/addgene:81165 ; RRID:Addgene_81165)

  • For your References section:

    Trade-offs between enzyme fitness and solubility illuminated by deep mutational scanning. Klesmith JR, Bacik JP, Wrenbeck EE, Michalczyk R, Whitehead TA. Proc Natl Acad Sci U S A. 2017 Feb 14. pii: 201614437. doi: 10.1073/pnas.1614437114. 10.1073/pnas.1614437114 PubMed 28196882
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