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AAV_Hs.TRAC_SEED_HDR_(TCRAFT2_makeTCR)
(Plasmid #256431)

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  • Purpose
    (Empty Backbone) CRISPR mediated HDR knock-in integration of TCRs into the Hs.TRAC locus using the SEED selection method, compatible with both makeTCR (BsmBI) and TCRAFT2 (SapI) high-throughput Golden Gate TCR cloning
  • Depositing Lab
    Michael Platten
  • Depositing Organization
    Deutsches Krebsforschungszentrum (DKFZ)
    Located in Germany
  • Publication
    Hamberger et al bioRxiv 2025.04.27.647198 ( How to cite )
  • Sequence Information

Ordering

This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 256431 Standard format: Plasmid sent in bacteria as agar stab 1 $94
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Backbone

  • Vector backbone
    pAAV2-Amp
  • Backbone manufacturer
    ELEGEN
  • Backbone size (bp) 2594
  • Modifications to backbone
    Removal of BsmBI sites in Amp promoter, removal of SapI sites, removal of lac promoter
  • Vector type
    Mammalian Expression, AAV
  • Promoter TRAC (knockin)
  • Selectable markers
    SEED selection - see https://www.nature.com/articles/s41587-024-02531-6

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    30°C
  • Growth Strain(s)
    NEB Stable
  • Growth instructions
    Recommend growing at low temperature in Recombination deficient bacteria due to the presence of repeat 141bp AAV2 LTR sequences
  • Copy number
    High Copy

Cloning Information

  • Cloning method Golden Gate

Resource Information

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry

Trademarks:

  • Zeocin® is an InvivoGen trademark.

Depositor Comments

Compatible with the Eyquem lab's SEED-Selection TCR knocking sorting technique when cloned with suitable TRBC domains (i.e. containing the D112K mutation), see https://www.nature.com/articles/s41587-024-02531-6

Compatible with BsmBI mediated makeTCR system, see https://www.biorxiv.org/content/10.1101/2025.04.27.647198v2

Compatible with Birnbaum lab's SapI mediated TCRAFT system, see https://linkinghub.elsevier.com/retrieve/pii/S1074761326000014

Replacement for previous version of vector (unreleased) that was not compatible with SEED selection sgRNA target site.

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:

    AAV_Hs.TRAC_SEED_HDR_(TCRAFT2_makeTCR) was a gift from Michael Platten (Addgene plasmid # 256431 ; http://n2t.net/addgene:256431 ; RRID:Addgene_256431)

  • For your References section:

    makeTCR: A Modular Platform for Rapid, Flexible, Scalable, Single-Step T Cell Receptor Synthesis. Hamberger M, Neuhoff M-T, Pietrantonio SV, Boschert T, Maldonado Torres C, Errerd A, Tan CL, Lindner JM, Platten M, Green EW. bioRxiv 2025.04.27.647198 10.1101/2025.04.27.647198
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