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N-Terminal Split Cas9 D10A Nickase with GyrA intein
(Plasmid #58695)


Item Catalog # Description Quantity Price (USD)
Plasmid 58695 Standard format: Plasmid sent in bacteria as agar stab 1 $85

This material is available to academics and nonprofits only.


  • Vector backbone
    Plasmid 42235: pX335-U6-Chimeric_BB-CBh-hSpCas9n(D10A)
  • Backbone size w/o insert (bp) 6412
  • Total vector size (bp) 6769
  • Modifications to backbone
    Inserted 5' ITR using Afl3 site in pX335 backbone as well as split the hSpCas9 D10A nickase using EcoRI/EcoRV and replaced the c-terminus with the GyrA Nsplit intein for protein trans-splicing with the C-Terminal Split Cas9 with GyrA intein plasmid
  • Vector type
    Mammalian Expression, AAV, CRISPR

Growth in Bacteria

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


  • Gene/Insert name
    D10A Nickase humanized S. pyogenes Cas9 with Gyra Nsplit Intein
  • Alt name
    GyraN_hSpCas9n (D10A)
  • Alt name
  • Species
    H. sapiens (human)
  • Insert Size (bp)
  • Mutation
    D10A nickase converting mutation to SpCas9
  • Promoter CBh
  • Tags / Fusion Proteins
    • GyrA Nsplit Intein (C terminal on insert)
    • NLS (N terminal on insert)
    • HA Tag (N terminal on insert)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site EcoRV (not destroyed)
  • 3′ cloning site EcoRI (not destroyed)
  • 5′ sequencing primer cctccgggctgtaattagc
  • 3′ sequencing primer AGGGTCAAGGAAGGCACG
  • (Common Sequencing Primers)

Resource Information

  • A portion of this plasmid was derived from a plasmid made by
    The Bao Lab modified the plasmids pX330 and pX335, which were received from AddGene (#42230 and #42335)and originally deposited by Feng Zhang

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry
  • 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:

    N-Terminal Split Cas9 D10A Nickase with GyrA intein was a gift from Gang Bao (Addgene plasmid # 58695 ; ; RRID:Addgene_58695)
  • For your References section:

    Trans-spliced Cas9 allows cleavage of HBB and CCR5 genes in human cells using compact expression cassettes. Fine EJ, Appleton CM, White DE, Brown MT, Deshmukh H, Kemp ML, Bao G. Sci Rep. 2015 Jul 1;5:10777. doi: 10.1038/srep10777. 10.1038/srep10777 PubMed 26126518