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CVS-N2c-RVdG-MCP:oScarlet:KASH-6xMBS
(Plasmid #231011)

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Item Catalog # Description Quantity Price (USD)
Plasmid 231011 Standard format: Plasmid sent in bacteria as agar stab 1 $89
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Backbone

  • Vector backbone
    Rabies CVS N2c(deltaG)
  • Backbone manufacturer
    Matthias J. Schnell, Thomas Jefferson University
  • Backbone size w/o insert (bp) 13959
  • Total vector size (bp) 15287
  • Modifications to backbone
    T7 promoter includes the ATAAT sequence at the 3' end for improved transcription (Conrad et al., 2020). HDVrz includes supercut "AGGCA" at the 3' end for improved cleavage efficiency (Ghanem et al., 2012). HDVrz is followed by a minimal WPRE element and SV40 polyA signal to promote nuclear export of CAG-driven genome during virus production. SV40 mammalian origin of replication was incorporated to promote plasmid replication in HEK293T. f1 phage-derived origin of replication was removed.
  • Vector type
    Neurotropic virus

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    30°C
  • Growth Strain(s)
    DH5alpha
  • Growth instructions
    Due to the size, this plasmid can behave like a low copy number plasmid in DH5alpha. If you experience difficulty, depositors had very good experience with Stellar cells from Takara, growing at 30 °C for 24 h.
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    MCP-oScarlet-KASH-6xMBS-SacIIMCS
  • Species
    Synthetic
  • Insert Size (bp)
    1705
  • Tags / Fusion Proteins
    • MCP (N terminal on insert)
    • KASH (C terminal on insert)

Cloning Information

  • Cloning method Gibson Cloning
  • 5′ sequencing primer aacatccctcaaaagacc
  • 3′ sequencing primer ctggatgagaagtgttgc
  • (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

Please visit https://doi.org/10.1101/2024.10.01.616167 for bioRxiv preprint.

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:

    CVS-N2c-RVdG-MCP:oScarlet:KASH-6xMBS was a gift from Tomasz Nowakowski (Addgene plasmid # 231011 ; http://n2t.net/addgene:231011 ; RRID:Addgene_231011)

  • For your References section:

    High-Complexity Barcoded Rabies Virus for Scalable Circuit Mapping Using Single-Cell and Single-Nucleus Sequencing. Shin D, Urbanek ME, Larson HH, Moussa AJ, Lee KY, Baker DL, Standen-Bloom E, Ramachandran S, Bogdanoff D, Cadwell CR, Nowakowski TJ. bioRxiv [Preprint]. 2024 Dec 11:2024.10.01.616167. doi: 10.1101/2024.10.01.616167. 10.1101/2024.10.01.616167 PubMed 39713304
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