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Addgene

pDB2-hCRY2-Luciferase-CD4-bla
(Plasmid #189985)

Ordering

This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 189985 Standard format: Plasmid sent in bacteria as agar stab 1 $85

Backbone

  • Vector backbone
    pDB2, derived from pUC57
  • Backbone size w/o insert (bp) 8417
  • Total vector size (bp) 10064
  • Modifications to backbone
    regions homologous to the hCRY2 genomic region (surrounding STOP codon) flank insert + floxed positive selection cassette (mCerulean-P2A-blaR). Silent mutations to the hCRY2 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 #189989 or #189990
  • 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
    Luciferase
  • Alt name
    fLUC
  • Species
    Firefly
  • Insert Size (bp)
    1647
  • Tag / Fusion Protein
    • 3xFlag (C terminal on insert)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site MluI (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

Terms and Licenses

Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

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:

    pDB2-hCRY2-Luciferase-CD4-bla was a gift from Achim Kramer (Addgene plasmid # 189985 ; http://n2t.net/addgene:189985 ; RRID:Addgene_189985)
  • For your References section:

    Circadian period is compensated for repressor protein turnover rates in single cells. Gabriel CH, Del Olmo M, Rizki Widini A, Roshanbin R, Woyde J, Hamza E, Gutu NN, Zehtabian A, Ewers H, Granada A, Herzel H, Kramer A. Proc Natl Acad Sci U S A. 2024 Aug 20;121(34):e2404738121. doi: 10.1073/pnas.2404738121. Epub 2024 Aug 14. 10.1073/pnas.2404738121 PubMed 39141353