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pKLV2-U6gRNA5(gBFP)-PGKmCherry2ABFP-W
(Plasmid #67986)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 67986 Standard format: Plasmid sent in bacteria as agar stab 1 $65

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pKLV2 lentiviral vector
  • Backbone size w/o insert (bp) 5783
  • Total vector size (bp) 8763
  • Vector type
    Lentiviral, CRISPR

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    Stbl3
  • Copy number
    High Copy

Gene/Insert 1

  • Gene/Insert name
    U6gRNA cassette, PGKmCherryABFP cassette, WPRE
  • Species
    H. sapiens (human), M. musculus (mouse)
  • Insert Size (bp)
    3005
  • Mutation
    Deleted BbsI site within WPRE, modified BFP

Cloning Information for Gene/Insert 1

  • Cloning method Restriction Enzyme
  • 5′ cloning site MluI (not destroyed)
  • 3′ cloning site KpnI (not destroyed)
  • 5′ sequencing primer CATAATAGCAACAGACATAC
  • (Common Sequencing Primers)

Gene/Insert 2

  • Gene/Insert name
    Guide RNA targeting modified BFP
  • Species
    Synthetic
  • Insert Size (bp)
    20

Cloning Information for Gene/Insert 2

  • Cloning method Restriction Enzyme
  • 5′ cloning site Bb (destroyed during cloning)
  • 3′ cloning site BbsI (destroyed during cloning)
  • 5′ sequencing primer GTAATAATTTCTTGGGTAGT
  • (Common Sequencing Primers)

Resource Information

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:

    pKLV2-U6gRNA5(gBFP)-PGKmCherry2ABFP-W was a gift from Kosuke Yusa (Addgene plasmid # 67986 ; http://n2t.net/addgene:67986 ; RRID:Addgene_67986)
  • For your References section:

    A CRISPR Dropout Screen Identifies Genetic Vulnerabilities and Therapeutic Targets in Acute Myeloid Leukemia. Tzelepis K, Koike-Yusa H, De Braekeleer E, Li Y, Metzakopian E, Dovey OM, Mupo A, Grinkevich V, Li M, Mazan M, Gozdecka M, Ohnishi S, Cooper J, Patel M, McKerrell T, Chen B, Domingues AF, Gallipoli P, Teichmann S, Ponstingl H, McDermott U, Saez-Rodriguez J, Huntly BJ, Iorio F, Pina C, Vassiliou GS, Yusa K. Cell Rep. 2016 Oct 18;17(4):1193-1205. doi: 10.1016/j.celrep.2016.09.079. 10.1016/j.celrep.2016.09.079 PubMed 27760321