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pKS diaCas9_sgRNA
(Plasmid #74923)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 74923 Standard format: Plasmid sent in bacteria as agar stab 1 $85

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pBlueScript KS(+)
  • Backbone manufacturer
    Stratagene
  • Backbone size w/o insert (bp) 2904
  • Total vector size (bp) 8577
  • Modifications to backbone
    Bsa1 site located in ampicillin resistance gene removed
  • Vector type
    CRISPR

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    High Copy

Gene/Insert 1

  • Gene/Insert name
    LHCF2 promoter
  • Species
    Phaeodactylum tricornutum
  • Insert Size (bp)
    442
  • GenBank ID
    Z24761.1
  • Promoter Cas9 module

Cloning Information for Gene/Insert 1

  • Cloning method Restriction Enzyme
  • 5′ cloning site Sac1 (not destroyed)
  • 3′ cloning site Xba1 (not destroyed)
  • 5′ sequencing primer none
  • 3′ sequencing primer none
  • (Common Sequencing Primers)

Gene/Insert 2

  • Gene/Insert name
    diaCas9
  • Species
    Synthetic
  • Insert Size (bp)
    4283
  • Promoter Cas9 module

Cloning Information for Gene/Insert 2

  • Cloning method Restriction Enzyme
  • 5′ cloning site Xba1 (not destroyed)
  • 3′ cloning site BamHI (not destroyed)
  • 5′ sequencing primer none
  • 3′ sequencing primer none
  • (Common Sequencing Primers)

Gene/Insert 3

  • Gene/Insert name
    LHCF1 terminator
  • Species
    Phaeodactylum tricornutum
  • Insert Size (bp)
    216
  • GenBank ID
    Z24761.1
  • Promoter Cas9 module

Cloning Information for Gene/Insert 3

  • Cloning method Restriction Enzyme
  • 5′ cloning site BamHI (not destroyed)
  • 3′ cloning site Pst1 (not destroyed)
  • 5′ sequencing primer none
  • 3′ sequencing primer none
  • (Common Sequencing Primers)

Gene/Insert 4

  • Gene/Insert name
    U6 promoter
  • Species
    Phaeodactylum tricornutum
  • Insert Size (bp)
    280
  • GenBank ID
    CM000611.1
  • Promoter sgRNA module

Cloning Information for Gene/Insert 4

  • Cloning method Restriction Enzyme
  • 5′ cloning site Pst1 (not destroyed)
  • 3′ cloning site none (unknown if destroyed)
  • 5′ sequencing primer none
  • 3′ sequencing primer none
  • (Common Sequencing Primers)

Gene/Insert 5

  • Gene/Insert name
    sgRNA
  • Species
    Synthetic
  • Insert Size (bp)
    103
  • Promoter sgRNA module

Cloning Information for Gene/Insert 5

Gene/Insert 6

  • Gene/Insert name
    U6 3' region
  • Species
    Phaeodactylum tricornutum
  • Insert Size (bp)
    330
  • GenBank ID
    CM000611.1
  • Promoter sgRNA module

Cloning Information for Gene/Insert 6

  • Cloning method Restriction Enzyme
  • 5′ cloning site none (unknown if destroyed)
  • 3′ cloning site HindIII (not destroyed)
  • 5′ sequencing primer none
  • 3′ sequencing primer none
  • (Common Sequencing Primers)

Resource Information

Terms and Licenses

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

Depositor Comments

The diaCas9 module was synthesized by GeneArt® Services Thermo Fisher Scientific Inc.
The sgRNA module was synthesized by Eurofins Genomic Services Ltd.

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:

    pKS diaCas9_sgRNA was a gift from Per Winge (Addgene plasmid # 74923 ; http://n2t.net/addgene:74923 ; RRID:Addgene_74923)
  • For your References section:

    A CRISPR/Cas9 system adapted for gene editing in marine algae. Nymark M, Sharma AK, Sparstad T, Bones AM, Winge P. Sci Rep. 2016 Apr 25;6:24951. doi: 10.1038/srep24951. 10.1038/srep24951 PubMed 27108533