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pAc-y1sgRNA-Cas9
(Plasmid #49331)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 49331 Plasmid sent as bacteria in agar stab 1 $65

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pAc-STABLE1-Puro
  • Backbone manufacturer
    Dr. James Sutherland
  • Backbone size w/o insert (bp) 6600
  • Total vector size (bp) 10658
  • Vector type
    Insect Expression, CRISPR
  • Selectable markers
    Puromycin

Growth in Bacteria

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

Gene/Insert 1

  • Gene/Insert name
    Cas9
  • Alt name
    CRISPR associated 9
  • Alt name
    hspCas9
  • Species
    Synthetic; Streptococcus pyogenes
  • Insert Size (bp)
    4278
  • Mutation
    Human codon optimised
  • Promoter Actin-5c
  • Tags / Fusion Proteins
    • 3xFLAG (N terminal on insert)
    • NLS (N terminal on insert)
    • NLS (C terminal on insert)

Cloning Information for Gene/Insert 1

  • Cloning method Restriction Enzyme
  • 5′ cloning site EcoRI (not destroyed)
  • 3′ cloning site HindIII (not destroyed)
  • 5′ sequencing primer gagttcttgtgctgtgtgga
  • 3′ sequencing primer tcgagacaaacggcgaaacc
  • (Common Sequencing Primers)

Gene/Insert 2

  • Gene/Insert name
    dU6-sgRNA
  • Species
    D. melanogaster (fly), Synthetic
  • Insert Size (bp)
    500
  • Promoter Drosophila U6

Cloning Information for Gene/Insert 2

  • Cloning method Restriction Enzyme
  • 5′ cloning site BglII (not destroyed)
  • 3′ cloning site BglII (not destroyed)
  • 5′ sequencing primer GTTCGACTTGCAGCCTGAAATACG
  • 3′ sequencing primer AAAAAAGCACCGACTCGGTGC
  • (Common Sequencing Primers)

Gene/Insert 3

  • Gene/Insert name
    y1 sgRNA
  • Species
    D. melanogaster (fly)
  • Insert Size (bp)
    20
  • Entrez Gene
    y (a.k.a. Dmel_CG3757, CG3757, Dmel\CG3757, EG:125H10.2, T6, Y)
  • Promoter Drosophila U6

Cloning Information for Gene/Insert 3

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

Resource Information

  • A portion of this plasmid was derived from a plasmid made by
    The human codon optimised spCas9 gene was amplified from pX330 (Addgene plasmid 42230) deposited by Feng Zhang. The pAc-STABLE1-Puro plasmid was obtained from Dr James Sutherland.
  • Terms and Licenses
  • Article Citing this Plasmid

Depositor Comments

gRNA target sequence GGTTTTGGACACTGGAACCG

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:

    pAc-y1sgRNA-Cas9 was a gift from Ji-Long Liu (Addgene plasmid # 49331)
  • For your References section:

    Mutagenesis and homologous recombination in Drosophila cell lines using CRISPR/Cas9. Bassett AR, Tibbit C, Ponting CP, Liu JL. Biol Open. 2013 Dec 10. pii: bio.20137120v1. doi: 10.1242/bio.20137120. 10.1242/bio.20137120 PubMed 24326186