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PB-U6-gRNA-Ef1a-CRISPRi-blast
(Plasmid #262893)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 262893 Standard format: Plasmid sent in bacteria as agar stab 1 $94
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DNA 262893-D50 50 μg of DNA in Tris buffer $495
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DNA 262893-D100 100 μg of DNA in Tris buffer $585
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Backbone

  • Vector backbone
    piggyBac vector
  • Backbone size w/o insert (bp) 4697
  • Total vector size (bp) 11281
  • Vector type
    Mammalian Expression, CRISPR ; piggyBac
  • Selectable markers
    Blasticidin

Growth in Bacteria

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

Gene/Insert 1

  • Gene/Insert name
    U6-gRNA Cloning Site (BsmB1 GoldenGate)
  • Species
    H. sapiens (human), Synthetic
  • Insert Size (bp)
    341
  • Promoter U6

Cloning Information for Gene/Insert 1

Gene/Insert 2

  • Gene/Insert name
    KRAB-dCas9-MeCP2
  • Insert Size (bp)
    5832
  • Promoter Ef1a
  • Tag / Fusion Protein
    • blasticidin S deaminase (C terminal on insert)

Cloning Information for Gene/Insert 2

Resource Information

  • A portion of this plasmid was derived from a plasmid made by
    iPSC Neurodegenerative Disease Initiative (iNDI) & Michael WardThe piggyBac vector was built from PB-TO-oNgn2 (Addgene plasmid # 198397); Neville Sanjana: The CRISPRi cassette came from lentiCRISPRi(v2)-Blast (Addgene plasmid #170068); Feng Zhang: The U6-gRNA cassette started with pSpCas9(BB)-2A-Puro (PX459) V2.0 (Addgene plasmid #62988), but the BbsI scaffold element was replaced with a new scaffold element including two BsmB1 sites for Golden Gate cloning of gRNA sequences.

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:

    PB-U6-gRNA-Ef1a-CRISPRi-blast was a gift from Ronald Hart (Addgene plasmid # 262893 ; http://n2t.net/addgene:262893 ; RRID:Addgene_262893)
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