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pET28a(+)_His6_Halo_TEV_SrcSH2pYsuperbinder
(Plasmid #260541)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 260541 Standard format: Plasmid sent in bacteria as agar stab 1 $94
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Backbone

  • Vector backbone
    pET28a(+)
  • Backbone manufacturer
    TWIST
  • Backbone size w/o insert (bp) 5233
  • Total vector size (bp) 6522
  • Modifications to backbone
    None
  • Vector type
    Bacterial Expression, Affinity Reagent/ Antibody

Growth in Bacteria

  • Bacterial Resistance(s)
    Kanamycin, 50 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Growth instructions
    Standard growth conditions used.
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    Src SH2 pY superbinder
  • Alt name
    sSrc
  • Alt name
    Chain A, Proto-oncogene tyrosine-protein kinase Src [Homo sapiens]
  • Species
    H. sapiens (human)
  • Insert Size (bp)
    327
  • Mutation
    Thr43Val, Cys48Ala, Lys66Leu as identified by original Kaneko et al Science Signaling 2012 study
  • Promoter lacIQ
  • Tags / Fusion Proteins
    • His6 (N terminal on insert)
    • HaloTag7 (N terminal on insert)
    • Tobacco Etch Virus Cleavage site (N terminal on insert)

Cloning Information

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

Resource Information

  • A portion of this plasmid was derived from a plasmid made by
    Src SH2 pY superbinder insert sequence was originally evolved by Tomonori Kaneko et al., Superbinder SH2 Domains Act as Antagonists of Cell Signaling. Sci. Signal.5, ra68-ra68 (2012). DOI:10.1126/scisignal.2003021 HaloTag7 sequence was based on: SnoopLigase Catalyzes Peptide-Peptide Locking and Enables Solid-Phase Conjugate Isolation. Buldun CM, Jean JX, Bedford MR, Howarth M. J Am Chem Soc. 2018 Feb 28;140(8):3008-3018. doi: 10.1021/jacs.7b13237. Epub 2018 Feb 14. 10.1021/jacs.7b13237 PubMed 29402082
  • Reference

Terms and Licenses

Trademarks:

  • Zeocin® is an InvivoGen trademark.

Depositor Comments

Accompanying protocol for phosphotyrosine peptide enrichment using HaloTag-sSrc can be found at: https://www.protocols.io/view/scalable-phosphotyrosine-enrichment-with-sh2-super-q26g7n3y9lwz/v1

HaloLink beads (magnetic or agarose) were purchased from Promega.

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:

    pET28a(+)_His6_Halo_TEV_SrcSH2pYsuperbinder was a gift from Judit Villén (Addgene plasmid # 260541 ; http://n2t.net/addgene:260541 ; RRID:Addgene_260541)

  • For your References section:

    Scalable phosphotyrosine enrichment with SH2 superbinder enables deep profiling of EGF responses. Chang AT, Rodriguez-Mias RA, Berg MD, Moggridge S, Villen J. EMBO J. 2026 Jul 3. doi: 10.1038/s44318-026-00843-8. 10.1038/s44318-026-00843-8 PubMed 42399519
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