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KI(GAPDH)-P2A-EGFP-CMV-NeoR
(Plasmid #114009)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 114009 Standard format: Plasmid sent in bacteria as agar stab 1 $85
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Backbone

  • Vector backbone
    LSL-Cas9-Rosa26TV
  • Backbone size w/o insert (bp) 4900
  • Total vector size (bp) 7350
  • Vector type
    Mammalian Expression
  • Selectable markers
    Neomycin (select with G418)

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    High Copy

Gene/Insert 1

  • Gene/Insert name
    GAPDH left arm
  • Species
    G. gallus (chicken)
  • Insert Size (bp)
    1000
  • GenBank ID
    NM_204305.1
  • Entrez Gene
    GAPDH (a.k.a. KNC-NDS6)

Cloning Information for Gene/Insert 1

  • Cloning method Restriction Enzyme
  • 5′ cloning site MauBI (not destroyed)
  • 3′ cloning site PmeI (not destroyed)
  • 5′ sequencing primer TTGTTGACCTGACCTGCCGTCTGGAG
  • 3′ sequencing primer CTCCTTGGATGCCATGTGGACCATCAAG
    • (Common Sequencing Primers)

Gene/Insert 2

  • Gene/Insert name
    eGFP
  • Insert Size (bp)
    700
  • Promoter endogenous promoter (chicken GAPDH in the case of insertion)

Cloning Information for Gene/Insert 2

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

Gene/Insert 3

  • Gene/Insert name
    NeoR
  • Insert Size (bp)
    800
  • Promoter CMV

Cloning Information for Gene/Insert 3

  • Cloning method Restriction Enzyme
  • 5′ cloning site NotI (not destroyed)
  • 3′ cloning site AscI (not destroyed)
  • 5′ sequencing primer ctagtggtgccagggcgtgc
  • 3′ sequencing primer ggcgcgcccTAAGATACATTG
    • (Common Sequencing Primers)

Gene/Insert 4

  • Gene/Insert name
    GAPDH right arm
  • Species
    G. gallus (chicken)
  • Insert Size (bp)
    3000
  • GenBank ID
    NM_204305.1
  • Promoter -

Cloning Information for Gene/Insert 4

  • Cloning method Restriction Enzyme
  • 5′ cloning site AscI (not destroyed)
  • 3′ cloning site SgrDI (not destroyed)
  • 5′ sequencing primer GCATTCTAGTTGTGGTTTGTCC
  • 3′ sequencing primer acccggtagaattcgatatcaagc
    • (Common Sequencing Primers)

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry
Trademarks:
  • Zeocin® is an InvivoGen trademark.
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:

    KI(GAPDH)-P2A-EGFP-CMV-NeoR was a gift from Pavel Volchkov (Addgene plasmid # 114009 ; http://n2t.net/addgene:114009 ; RRID:Addgene_114009)

  • For your References section:

    Successful CRISPR/Cas9 mediated homologous recombination in a chicken cell line. Antonova E, Glazova O, Gaponova A, Eremyan A, Zvereva S, Grebenkina N, Volkova N, Volchkov P. F1000Res. 2018 Feb 28;7:238. doi: 10.12688/f1000research.13457.2. eCollection 2018. 10.12688/f1000research.13457.2 PubMed 29946437
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