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LentiCRISPRv2-mCherry
(Plasmid #99154)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 99154 Standard format: Plasmid sent in bacteria as agar stab 1 $65

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    LentiCRISPRv2
  • Backbone manufacturer
    Feng Zhang
  • Backbone size w/o insert (bp) 10000
  • Total vector size (bp) 14984
  • Modifications to backbone
    mCherry P2A-linked to hSpCAS9 by InFusion cloning.
  • Vector type
    Mammalian Expression, Lentiviral, CRISPR
  • Selectable markers
    mCherry

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    NEB Stable
  • Growth instructions
    Use SapI digest to check for unwanted recombination of lentiviral plasmid. Only amplify in RecA- bacteria (eg. Stbl3).
  • Copy number
    High Copy

Gene/Insert 1

  • Gene/Insert name
    Cas9
  • Alt name
    S. pyogenes CRISPR-Cas9
  • Species
    Synthetic
  • Insert Size (bp)
    4200
  • Promoter EFS-NS
  • Tag / Fusion Protein
    • FLAG (C terminal on insert)

Cloning Information for Gene/Insert 1

  • Cloning method Restriction Enzyme
  • 5′ cloning site AgeI, XbaI, AfeI (not destroyed)
  • 3′ cloning site BamHI (not destroyed)
  • 5′ sequencing primer hEF1aprom-F
  • 3′ sequencing primer WPRE-R
  • (Common Sequencing Primers)

Gene/Insert 2

  • Gene/Insert name
    mCherry
  • Species
    Discoma sp.
  • Insert Size (bp)
    708
  • Promoter EFS-NS

Cloning Information for Gene/Insert 2

  • Cloning method Ligation Independent Cloning
  • 5′ sequencing primer ACC AAA GAG GTG CTG GAC G
  • 3′ sequencing primer WPRE-R
  • (Common Sequencing Primers)

Resource Information

Depositor Comments

This LentiCRISPRv2 variant was produced by restriction digest and multi-fragment InFusion (Clontech) assembly. It has been validated for cutting efficiency following transduction and FACS-sorting for mCherry. Suitable for single- and multi-colour guided CRISPR editing in stem cells and cell lines.

sgRNA of choice can be cloned in using BsmBI digest. Follow original LentiCRISPRv2 cloning protocol.

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:

    LentiCRISPRv2-mCherry was a gift from Agata Smogorzewska (Addgene plasmid # 99154 ; http://n2t.net/addgene:99154 ; RRID:Addgene_99154)