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pST1374-NLS-flag-linker-Cas9
(Plasmid #44758)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 44758 Standard format: Plasmid sent in bacteria as agar stab 1 $94
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DNA 44758-D50 50 μg of DNA in Tris buffer $495
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DNA 44758-D100 100 μg of DNA in Tris buffer $585
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Backbone

  • Vector backbone
    pST1374 (Addgene #13426)
  • Backbone manufacturer
    pST1374-based custom vector
  • Backbone size w/o insert (bp) 5052
  • Total vector size (bp) 9318
  • Vector type
    Mammalian Expression, 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
    Human codon optimized Cas9
  • Insert Size (bp)
    4266
  • Mutation
    codon-optimized
  • Promoter CMV
  • Tags / Fusion Proteins
    • Flag (N terminal on insert)
    • NLS (N terminal on insert)
    • 32 amino acid linker (N terminal on insert)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site NheI (not destroyed)
  • 3′ cloning site ApaI (not destroyed)
  • 5′ sequencing primer CMV-fwd
  • 3′ sequencing primer BGH-rev
  • (Common Sequencing Primers)

Terms and Licenses

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

Trademarks:

  • Zeocin® is an InvivoGen trademark.

Depositor Comments

The 32 amino acid linker sequence comes after the Flag tag and before the beginning of the Cas9 sequence. This 32 amino acid linker sequence is not depicted in the depositor's full plasmid sequence.

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

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

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

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

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:

    pST1374-NLS-flag-linker-Cas9 was a gift from Xingxu Huang (Addgene plasmid # 44758 ; http://n2t.net/addgene:44758 ; RRID:Addgene_44758)

  • For your References section:

    Generation of gene-modified mice via Cas9/RNA-mediated gene targeting. Shen B, Zhang J, Wu H, Wang J, Ma K, Li Z, Zhang X, Zhang P, Huang X. Cell Res. 2013 Apr 2. doi: 10.1038/cr.2013.46. 10.1038/cr.2013.46 PubMed 23545779
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