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pDB-hCRY1-Halo-CD4-bla
(Plasmid #179445)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 179445 Standard format: Plasmid sent in bacteria as agar stab 1 $94
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DNA 179445-D50 50 μg of DNA in Tris buffer $495
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DNA 179445-D100 100 μg of DNA in Tris buffer $585
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Backbone

  • Vector backbone
    pDB, derived from pUC57
  • Backbone manufacturer
    none
  • Backbone size w/o insert (bp) 8200
  • Total vector size (bp) 9122
  • 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
    HaloTag
  • Alt name
    Halo7
  • Species
    Synthetic
  • Insert Size (bp)
    887
  • Entrez Gene
    Halo7 (a.k.a. Halo7)
  • 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

  • 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.

Purpose

Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.

Delivery

  • Amount 50 μg
  • Concentration 1 μg/μL
  • Pricing $495 USD
  • Storage 4 ℃ (short-term) or -20 ℃ (long-term)

Terms and Licenses

Quality Control

Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.

Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.

Purpose

Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.

Delivery

  • Amount 100 μg
  • Concentration 1 μg/μL
  • Pricing $585 USD
  • Storage 4 ℃ (short-term) or -20 ℃ (long-term)

Terms and Licenses

Quality Control

Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.

Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.

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-Halo-CD4-bla was a gift from Achim Kramer (Addgene plasmid # 179445 ; http://n2t.net/addgene:179445 ; RRID:Addgene_179445)

  • 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
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