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pHL-EF1a-SphcCas9(D10A)-iP-A
(Plasmid #60600)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 60600 Standard format: Plasmid sent in bacteria as agar stab 1 $94
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DNA 60600-D50 50 μg of DNA in Tris buffer $495
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DNA 60600-D100 100 μg of DNA in Tris buffer $585
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Backbone

  • Vector backbone
    pHL-EF1a-GW-iP-A
  • Backbone manufacturer
    Hotta Lab, CiRA
  • Backbone size w/o insert (bp) 6923
  • Total vector size (bp) 9442
  • Modifications to backbone
    Gateway system (attR1/R2) compatible mammalian expression plasmid vector, especially optimized for the use in human iPS cells. Human EF1a promoter allows to express a gene of interest at the high-level in human pluripotent stem cells.
  • Vector type
    Mammalian Expression, CRISPR
  • Selectable markers
    Puromycin

Growth in Bacteria

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

Gene/Insert 1

  • Gene/Insert name
    CRISPR Cas9 D10A
  • Alt name
    SphcCas9 (D10A)
  • Species
    Streptococcus pyogenes
  • Insert Size (bp)
    4140
  • Mutation
    Changed Asp 10 to Ala, Codon usage optimized for human usage

Cloning Information for Gene/Insert 1

Gene/Insert 2

  • Gene/Insert name
    puromycin resistance gene
  • Alt name
    PuroR
  • Species
    Streptomyces alboniger
  • Insert Size (bp)
    600

Cloning Information for Gene/Insert 2

Terms and Licenses

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

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

  • 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:

    pHL-EF1a-SphcCas9(D10A)-iP-A was a gift from Akitsu Hotta (Addgene plasmid # 60600 ; http://n2t.net/addgene:60600 ; RRID:Addgene_60600)

  • For your References section:

    Precise Correction of the Dystrophin Gene in Duchenne Muscular Dystrophy Patient Induced Pluripotent Stem Cells by TALEN and CRISPR-Cas9. Li HL, Fujimoto N, Sasakawa N, Shirai S, Ohkame T, Sakuma T, Tanaka M, Amano N, Watanabe A, Sakurai H, Yamamoto T, Yamanaka S, Hotta A. Stem Cell Reports. 2014 Nov 25. pii: S2213-6711(14)00335-X. doi: 10.1016/j.stemcr.2014.10.013. 10.1016/j.stemcr.2014.10.013 PubMed 25434822
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