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pSpCas9(BB)-2A-Puro(PX459)-GJA1 sgRNA
(Plasmid #164471)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 164471 Standard format: Plasmid sent in bacteria as agar stab 1 $85
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Backbone

  • Vector backbone
    pSpCas9(BB)-2A-Puro (PX459)
  • Backbone manufacturer
    Feng Zhang
  • Backbone size w/o insert (bp) 9200
  • Total vector size (bp) 9220
  • Modifications to backbone
    Insertion of a sgRNA sequence targeting GJA1
  • Vector type
    Mammalian Expression
  • Selectable markers
    Puromycin

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
    Gap junction alpha-1 (GJA1)
  • Alt name
    CX43
  • gRNA/shRNA sequence
    AAGCCTACTCAACTGCTGGA
  • Species
    H. sapiens (human)
  • GenBank ID
    NM_000165.5
  • Entrez Gene
    GJA1 (a.k.a. AVSD3, CMDR, CX43, EKVP, EKVP3, GJAL, HLHS1, HSS, ODDD, PPKCA)
  • Promoter U6

Cloning Information

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

Resource Information

  • A portion of this plasmid was derived from a plasmid made by
    Matthew R. Cring

Terms and Licenses

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

Depositor Comments

A student in University of Iowa did this cloning.

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:

    pSpCas9(BB)-2A-Puro(PX459)-GJA1 sgRNA was a gift from Markus Kuehn (Addgene plasmid # 164471 ; http://n2t.net/addgene:164471 ; RRID:Addgene_164471)

  • For your References section:

    Absence of Connexin 43 results in smaller retinas and arrested, depolarized retinal progenitor cells in human retinal organoids. Cheng L, Cring MR, Wadkins DA, Kuehn MH. Stem Cells. 2022 Mar 9. pii: 6545973. doi: 10.1093/stmcls/sxac017. 10.1093/stmcls/sxac017 PubMed 35263762
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