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pMLM3705
(Plasmid #47754)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 47754 Standard format: Plasmid sent in bacteria as agar stab 1 $94
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DNA 47754-D50 50 μg of DNA in Tris buffer $495
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DNA 47754-D100 100 μg of DNA in Tris buffer $585
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Backbone

  • Vector backbone
    pJDS246
  • Backbone manufacturer
    Joung lab (Addgene plasmid # 43861)
  • Modifications to backbone
    codon-optimized Cas9 modified by site-directed mutagenesis to dCas9; added VP64 activation domain
  • Vector type
    Mammalian Expression, CRISPR

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    XL1 Blue
  • Copy number
    Unknown

Gene/Insert

  • Gene/Insert name
    codon optimized dCas9-VP64
  • Alt name
    inactive Cas9 nuclease
  • Alt name
    VP64 activation domain
  • Species
    Synthetic; Streptococcus pyogenes
  • Mutation
    D10A and H840A mutations in Cas9 (catalytically inactive)
  • Promoter CMV
  • Tags / Fusion Proteins
    • NLS (C terminal on insert)
    • 3x FLAG (C terminal on insert)
    • VP64 (C terminal on insert)

Cloning Information

Terms and Licenses

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

Trademarks:

  • Zeocin® is an InvivoGen trademark.

Depositor Comments

Plasmid Features:
Cas9 = 409-4512
NLS = 4519-4539
3X FLAG = 4546-4611
VP64 = 4633-4794

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:

    pMLM3705 was a gift from Keith Joung (Addgene plasmid # 47754 ; http://n2t.net/addgene:47754 ; RRID:Addgene_47754)

  • For your References section:

    CRISPR RNA-guided activation of endogenous human genes. Maeder ML, Linder SJ, Cascio VM, Fu Y, Ho QH, Joung JK. Nat Methods. 2013 Jul 25. doi: 10.1038/nmeth.2598. 10.1038/nmeth.2598 PubMed 23892898
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