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pBAD_GFP11_T7LysH17A
(Plasmid #59591)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 59591 Standard format: Plasmid sent in bacteria as agar stab 1 $85

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pBAD-MCS
  • Backbone manufacturer
    Arie Geerlof - EMBL
  • Backbone size (bp) 4332
  • Modifications to backbone
    The XhoI site of pBAD-MCS was first removed by site directed mutagenesis. Between the NcoI and HindIII sites of the modified pBAD-MCS vector a 132 bp DNA cassette was inserted for expression of cloned protein fragments with an N-terminal 6xHis-tag and a C-terminal GFP11 tag. Protein fragments can be cloned into this site using sticky end (SpeI and XhoI) or blunt end (PvuII) ligation. A 636 bp fragment of the plasmid pLysS, encompassing the T7 lysozyme gene, was ligated into a SphI site in a non-essential region of the vector. Finally the mutation H17A was introduced to the T7 lysozyme gene to reduce its amidase activity.
  • Vector type
    Bacterial Expression
  • Promoter pARA
  • Tags / Fusion Proteins
    • 6xHis-tag (N terminal on backbone)
    • GFP11 (C terminal on backbone)

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    Unknown

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ sequencing primer ATG CCA TAG CAT TTT TAT CC
  • 3′ sequencing primer GAT TTA ATC TGT ATC AGG
  • (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:

    pBAD_GFP11_T7LysH17A was a gift from Richard Kingston (Addgene plasmid # 59591 ; http://n2t.net/addgene:59591 ; RRID:Addgene_59591)
  • For your References section:

    Identifying protein domains by global analysis of soluble fragment data. Bulloch EM, Kingston RL. Anal Biochem. 2014 Jul 10. pii: S0003-2697(14)00280-2. doi: 10.1016/j.ab.2014.06.021. 10.1016/j.ab.2014.06.021 PubMed 25016192