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PB_tre_dCas9_KRAB
(Plasmid #126030)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 126030 Standard format: Plasmid sent in bacteria as agar stab 1 $94
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DNA 126030-D50 50 μg of DNA in Tris buffer $495
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DNA 126030-D100 100 μg of DNA in Tris buffer $585
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Backbone

  • Vector backbone
    piggyBac cargo vector
  • Backbone manufacturer
    System Biosciences
  • Total vector size (bp) 11669
  • Vector type
    Mammalian Expression, CRISPR ; piggyBac
  • Selectable markers
    Hygromycin

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    30°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    Unknown

Gene/Insert 1

  • Gene/Insert name
    dCas9 fusion with KRAB
  • Alt name
    dCas9-KRAB
  • Insert Size (bp)
    4460
  • Mutation
    D10A, H840A (catalytically inactive Cas9)
  • Promoter TRE

Cloning Information for Gene/Insert 1

Gene/Insert 2

  • Gene/Insert name
    Hygromycin resistance
  • Insert Size (bp)
    1008
  • Promoter EF1 alpha

Cloning Information for Gene/Insert 2

  • Cloning method Restriction Enzyme
  • 5′ cloning site HpaI (destroyed during cloning)
  • 3′ cloning site SpeI (destroyed during cloning)
  • 5′ sequencing primer NA
  • (Common Sequencing Primers)

Resource Information

  • A portion of this plasmid was derived from a plasmid made by
    dCas9-KRAB from Addgene #50917 (Kearns et al., 2014)

Terms and Licenses

Trademarks:

  • Zeocin® is an InvivoGen trademark.

Depositor Comments

Please visit https://www.biorxiv.org/content/10.1101/448803v1 for BioRxiv preprint

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:

    PB_tre_dCas9_KRAB was a gift from Mauro Calabrese (Addgene plasmid # 126030 ; http://n2t.net/addgene:126030 ; RRID:Addgene_126030)

  • For your References section:

    A piggyBac-based toolkit for inducible genome editing in mammalian cells. Schertzer MD, Thulson E, Braceros KCA, Lee DM, Hinkle ER, Murphy RM, Kim SO, Vitucci EM, Calabrese JM. RNA. 2019 May 17. pii: rna.068932.118. doi: 10.1261/rna.068932.118. 10.1261/rna.068932.118 PubMed 31101683
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