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pET30Xa-SBL1(160E,NoCys)
(Plasmid #66875)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 66875 Standard format: Plasmid sent in bacteria as agar stab 1 $85
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Backbone

  • Vector backbone
    pET30Xa-LIC
  • Backbone manufacturer
    Novagen
  • Backbone size w/o insert (bp) 5444
  • Total vector size (bp) 7978
  • Modifications to backbone
    11 bp C-term extension: TAAACTAGTCATGGGGC
  • Vector type
    Bacterial Expression

Growth in Bacteria

  • Bacterial Resistance(s)
    Kanamycin, 50 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Growth instructions
    37C before induction, 30 deg for 14-16+ hrs after induction
  • Copy number
    Unknown

Gene/Insert

  • Gene/Insert name
    soybean lipoxygenase-1 (S160E) with all Cys to Ser mutations
  • Species
    Glycine max
  • Insert Size (bp)
    2517
  • Mutation
    S160E; changed 4 (all) cysteines to serine
  • Promoter T7lac
  • Tag / Fusion Protein
    • Histag (N terminal on backbone)

Cloning Information

  • Cloning method Ligation Independent Cloning
  • 5′ sequencing primer tca tca ttc ttc tgg tct gg
  • 3′ sequencing primer ttg ccg act ctc att agc c
    • (Common Sequencing Primers)

Resource Information

Terms and Licenses

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

Depositor Comments

prepared by four sequential mutations of Cys to Ser, from pET30Xa-SBL1(160S,NoCys)

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:

    pET30Xa-SBL1(160E,NoCys) was a gift from Betty Gaffney (Addgene plasmid # 66875 ; http://n2t.net/addgene:66875 ; RRID:Addgene_66875)

  • For your References section:

    On the relationships of substrate orientation, hydrogen abstraction, and product stereochemistry in single and double dioxygenations by soybean lipoxygenase-1 and its Ala542Gly mutant. Coffa G, Imber AN, Maguire BC, Laxmikanthan G, Schneider C, Gaffney BJ, Brash AR. J Biol Chem. 2005 Nov 18;280(46):38756-66. Epub 2005 Sep 12. 10.1074/jbc.M504870200 PubMed 16157595
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