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pET45b-tsr(B/X) (6X-His-Dm-Cofilin)
(Plasmid #128278)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 128278 Standard format: Plasmid sent in bacteria as agar stab 1 $85
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Backbone

  • Vector backbone
    pet45b(+)
  • Backbone manufacturer
    Novagen
  • Backbone size w/o insert (bp) 5260
  • Total vector size (bp) 5650
  • Vector type
    Bacterial Expression

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Growth instructions
    Grow with shaking at 225 rpm in Luria broth or 2X YT broth. For optimizing expression of the protein of interest, use appropriate E. coli protein expression strain such as BL21 (DE3) cells in 2X YT broth. Plasmid will be maintained at lower copy in these strains than in chemically-competent cloning strains such as E. coli Top10(TM) or DH10B(TM).
  • Copy number
    Low Copy

Gene/Insert

  • Gene/Insert name
    twinstar (tsr)
  • Alt name
    Dm-Cofilin
  • Species
    D. melanogaster (fly)
  • Insert Size (bp)
    452
  • Mutation
    Met1 changed to Pro1; Cys77 silent basepair mutation/codon switch from UGU to UGC
  • GenBank ID
    U24490.1 U24676.2
  • Entrez Gene
    tsr (a.k.a. Dmel_CG4254, 12/8, ADF, ADF/cofilin, Adf, CG4254, Cadf, D-cof, D61, Dmel\CG4254, T8, Tsr, cofilin, cofilin/ADF, cofilin/actin depolymerizing factor homolog, l(2)03101, l(2)k03101, l(2)k05633, l(2)k13409, ntf)
  • Promoter T7
  • Tag / Fusion Protein
    • 6X Histidine tag (N terminal on insert)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site BamHI (not destroyed)
  • 3′ cloning site XhoI (not destroyed)
  • 5′ sequencing primer T7 (Eurofins standard primers)
  • 3′ sequencing primer T7 terminal (Eurofins standard primers)
    • (Common Sequencing Primers)

Resource Information

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:

    pET45b-tsr(B/X) (6X-His-Dm-Cofilin) was a gift from Anna Sokac (Addgene plasmid # 128278 ; http://n2t.net/addgene:128278 ; RRID:Addgene_128278)

  • For your References section:

    Cofilin-Mediated Actin Stress Response Is Maladaptive in Heat-Stressed Embryos. Figard L, Zheng L, Biel N, Xue Z, Seede H, Coleman S, Golding I, Sokac AM. Cell Rep. 2019 Mar 26;26(13):3493-3501.e4. doi: 10.1016/j.celrep.2019.02.092. 10.1016/j.celrep.2019.02.092 PubMed 30917306
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