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Addgene

pDB-hCRY1-mClover3-CD4-bla
(Plasmid #179441)

Ordering

This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 179441 Standard format: Plasmid sent in bacteria as agar stab 1 $85

Backbone

  • Vector backbone
    pDB, derived from pUC57
  • Backbone manufacturer
    none
  • Backbone size w/o insert (bp) 8200
  • Total vector size (bp) 8948
  • Modifications to backbone
    regions homologous to the hCRY1 genomic region (surrounding STOP codon) flank insert+positive selection cassette (mCerulean-P2A-blaR). Silent mutations to the hCRY1 genomic sequence were inserted to avoid donor vector targeting by Cas9/sgRNA. Negative Selection cassette expresses hCD4. Should be used together with Cas9/sgRNA plasmid #179453
  • Vector type
    CRISPR
  • Selectable markers
    Blasticidin

Growth in Bacteria

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

Gene/Insert

  • Gene/Insert name
    mClover3
  • Species
    Aequorea victoria
  • Insert Size (bp)
    714
  • Promoter None
  • Tag / Fusion Protein
    • His/Flag (C terminal on insert)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site AgeI (not destroyed)
  • 3′ cloning site SpeI (not destroyed)
  • 5′ sequencing primer --
  • 3′ sequencing primer --
  • (Common Sequencing Primers)

Resource Information

  • Supplemental Documents
  • A portion of this plasmid was derived from a plasmid made by
    hCD4 was cloned from Addgene #35712, mClover3 was cloned from #74252

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:

    pDB-hCRY1-mClover3-CD4-bla was a gift from Achim Kramer (Addgene plasmid # 179441 ; http://n2t.net/addgene:179441 ; RRID:Addgene_179441)
  • For your References section:

    Live-cell imaging of circadian clock protein dynamics in CRISPR-generated knock-in cells. Gabriel CH, Del Olmo M, Zehtabian A, Jager M, Reischl S, van Dijk H, Ulbricht C, Rakhymzhan A, Korte T, Koller B, Grudziecki A, Maier B, Herrmann A, Niesner R, Zemojtel T, Ewers H, Granada AE, Herzel H, Kramer A. Nat Commun. 2021 Jun 18;12(1):3796. doi: 10.1038/s41467-021-24086-9. 10.1038/s41467-021-24086-9 PubMed 34145278