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C-Terminal Split Cas9 with GyrA intein
(Plasmid #58694)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 58694 Standard format: Plasmid sent in bacteria as agar stab 1 $65

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    Plasmid 42230: pX330-U6-Chimeric_BB-CBh-hSpCas9
  • Backbone size w/o insert (bp) 6580
  • Total vector size (bp) 6812
  • Modifications to backbone
    Inserted 5' ITR using Afl3 cloning site of pX330 vector and split hSpCas9 using AgeI/PmlI to replace 5' portion of hSpCas9 with Gyra Csplit intein that when combined with the GyraNsplit intein backbone splices the two components together forming full length hSpCas9
  • Vector type
    Mammalian Expression, AAV, CRISPR

Growth in Bacteria

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

Gene/Insert

  • Gene/Insert name
    Humanized C-Terminal S. pyogenes Cas9 with GyrA Csplit Intein
  • Alt name
    GyraC_hSpCas9
  • Alt name
    tsCas9-C
  • Species
    H. sapiens (human)
  • Insert Size (bp)
    237
  • Promoter CBh
  • Tags / Fusion Proteins
    • GyrA Csplit Intein (N terminal on insert)
    • NLS (C terminal on insert)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site AgeI (not destroyed)
  • 3′ cloning site PmlI (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 plasmids pX330 and pX335, which were received from AddGene (#42230 and #42335)and originally deposited by Feng Zhang
  • Terms and Licenses
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:

    C-Terminal Split Cas9 with GyrA intein was a gift from Gang Bao (Addgene plasmid # 58694 ; http://n2t.net/addgene:58694 ; RRID:Addgene_58694)
  • 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