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PX458_3XHA_den1FnCas9_KRAB_T2A_eGFP
(Plasmid #200910)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 200910 Standard format: Plasmid sent in bacteria as agar stab 1 $94
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DNA 200910-D50 50 μg of DNA in Tris buffer $495
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DNA 200910-D100 100 μg of DNA in Tris buffer $585
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Backbone

  • Vector backbone
    PX458
  • Backbone size w/o insert (bp) 4832
  • Total vector size (bp) 10320
  • Vector type
    Mammalian Expression, CRISPR

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
    3XHA_den1FnCas9_KRAB_eGFP
  • Alt name
    Nuclease-dead en1FnCas9 with 3XHA tag, KRAB domain and eGFP for mammalian expression
  • Alt name
    en1FnCas9 is FnCas9b with E1369R substitution
  • Species
    Synthetic; Francisella novicida U112
  • Insert Size (bp)
    5488
  • Mutation
    Changed Glutamic Acid to Arginine at 1369 position
  • GenBank ID
    NZ_CP009607.1
  • Promoter CBH
  • Tags / Fusion Proteins
    • 3xHA-SV40NLS (N terminal on insert)
    • Nucleoplasmin NLS-KRAB-T2A-EGFP (C terminal on insert)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site BamH1 (not destroyed)
  • 3′ cloning site Eco53kI (not destroyed)
  • 5′ sequencing primer CTGGCTTATAGGTCTAGGCGCGTGAAGATCAAAAG
  • 3′ sequencing primer CTTTTGATCTTCACGCGCCTAGACCTATAAGCCAG
  • (Common Sequencing Primers)

Resource Information

  • A portion of this plasmid was derived from a plasmid made by
    Subcloned from PX458, a gift from Feng Zhang's Lab

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:

    PX458_3XHA_den1FnCas9_KRAB_T2A_eGFP was a gift from Debojyoti Chakraborty (Addgene plasmid # 200910 ; http://n2t.net/addgene:200910 ; RRID:Addgene_200910)
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