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Addgene

pLY74
(Plasmid #130945)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 130945 Standard format: Plasmid sent in bacteria as agar stab 1 $85
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Backbone

  • Vector backbone
    p15AC
  • Backbone manufacturer
    p15A vector from pdCas9-bacteria (Plasmid #44249)
  • Backbone size w/o insert (bp) 1725
  • Total vector size (bp) 3006
  • Modifications to backbone
    Adding 2 BbsI sites for construction
  • Vector type
    Bacterial Expression, Synthetic Biology

Growth in Bacteria

  • Bacterial Resistance(s)
    Chloramphenicol, 25 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Growth instructions
    Culture in the Lennox’s Luria-Bertani (LB) medium (10 g/L peptone, 5 g/L yeast extract, 5 g/L NaCl)
  • Copy number
    Low Copy

Gene/Insert

  • Gene/Insert name
    sgRNA-G6
  • gRNA/shRNA sequence
    AAAGCTGTTGTGGCAAAAAA
  • Species
    Synthetic
  • Promoter Plux2

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site EcoRI (destroyed during cloning)
  • 3′ cloning site SpeI (destroyed during cloning)
  • 5′ sequencing primer AACGGTCTGGTTATAGGTACATTGAGCAAC
  • 3′ sequencing primer GTTCGTAAGCCATTTCCGCTCGCCGCAGTC
    • (Common Sequencing Primers)

Terms and Licenses

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

Depositor Comments

The Top10 bacterial strain should be used for downstream applications

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:

    pLY74 was a gift from Baojun Wang (Addgene plasmid # 130945 ; http://n2t.net/addgene:130945 ; RRID:Addgene_130945)

  • For your References section:

    Engineered CRISPRa enables programmable eukaryote-like gene activation in bacteria. Liu Y, Wan X, Wang B. Nat Commun. 2019 Aug 26;10(1):3693. doi: 10.1038/s41467-019-11479-0. 10.1038/s41467-019-11479-0 PubMed 31451697
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