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pAH243
(Plasmid #121147)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 121147 Standard format: Plasmid sent in bacteria as agar stab 1 $75

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pSLF273
  • Backbone size w/o insert (bp) 8585
  • Total vector size (bp) 15174
  • Vector type
    Yeast Expression, CRISPR
  • Selectable markers
    S.pombe ura4

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    High Copy

Gene/Insert 1

  • Gene/Insert name
    humanized Streptococcus pyogenes Cas9
  • Species
    S. pombe (fission yeast), Synthetic
  • Insert Size (bp)
    5725
  • Promoter nmt41
  • Tag / Fusion Protein
    • Flag (N terminal on insert)

Cloning Information for Gene/Insert 1

  • Cloning method Restriction Enzyme
  • 5′ cloning site XhoI (destroyed during cloning)
  • 3′ cloning site BglII (not destroyed)
  • 5′ sequencing primer TCTCACTTTCTGACTTATAGTCGCT
  • 3′ sequencing primer AGCAGTACTGGCAAGGGAGAC
  • (Common Sequencing Primers)

Gene/Insert 2

  • Gene/Insert name
    gRNA expression module
  • Species
    S. pombe (fission yeast)
  • Promoter S.pombe rrk1

Cloning Information for Gene/Insert 2

  • Cloning method Restriction Enzyme
  • 5′ cloning site MluI (destroyed during cloning)
  • 3′ cloning site MluI (destroyed during cloning)
  • 5′ sequencing primer CACACATGAACAAGGAAGTACAGG
  • 3′ sequencing primer CAGATAAGTCACTATGTCCGAGTG
  • (Common Sequencing Primers)

Resource Information

Depositor Comments

Ran, F.A., Hsu, P.D.P., Wright, J., Agarwala, V., Scott, D. a and Zhang, F. (2013) Genome engineering using the CRISPR-Cas9 system. Nat. Protoc., 8, 2281–2308.

Jacobs, J.Z., Ciccaglione, K.M., Tournier, V. and Zaratiegui, M. (2014) Implementation of the CRISPR-Cas9 system in fission yeast. Nat. Commun., 5, 5344. After prepping the DNA, it is recommended to heat the DNA to 65C before digesting with Bbs I. Supplementary data is available at Figshare: https://doi.org/10.25387/g3.7685642.

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:

    pAH243 was a gift from Katsunori Tanaka (Addgene plasmid # 121147 ; http://n2t.net/addgene:121147 ; RRID:Addgene_121147)
  • For your References section:

    Short-Homology-Mediated CRISPR/Cas9-Based Method for Genome Editing in Fission Yeast. Hayashi A, Tanaka K. G3 (Bethesda). 2019 Feb 12. pii: g3.118.200976. doi: 10.1534/g3.118.200976. 10.1534/g3.118.200976 PubMed 30755408