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pAH-CTX1-rhadCas9
(Plasmid #129391)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 129391 Standard format: Plasmid sent in bacteria as agar stab 1 $75

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pAH-CTX1-rha
  • Backbone manufacturer
    Andrew Hogan
  • Backbone size w/o insert (bp) 7636
  • Total vector size (bp) 11897
  • Modifications to backbone
    Insertion of codon-optimized dCas9 for Burkholderia cenocepacia.
  • Vector type
    Bacterial Expression, CRISPR

Growth in Bacteria

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

Gene/Insert

  • Gene/Insert name
    Burkholderia cenocepacia codon-optimized dCas9
  • Species
    Synthetic; S. pyogenes
  • Insert Size (bp)
    4247
  • Mutation
    Entire dCas9 codon optimized for the GC-rich B. cenocepacia. See this sequence and that of pAH-CTX1-rhadCas9-native for comparison.
  • GenBank ID
    AE004092.2
  • Promoter PrhaBAD

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site NotI (not destroyed)
  • 3′ cloning site SpeI (not destroyed)
  • 5′ sequencing primer ATTTTTGCGGCGCGTGTAG
  • 3′ sequencing primer GGGTTCTATCGCCACGGAC
  • (Common Sequencing Primers)

Resource Information

  • A portion of this plasmid was derived from a plasmid made by
    Codon-optimized dCas9 was synthesized in two parts by IDT. Using ligation and PCR amplification, the full dCas9 vector was constructed and subsequently cloned into the pAH-CTX1-rha background. This insert was designed based on the dCas9 CDS from pdCas9-bacteria (Qi et al. 2013).
  • Terms and Licenses
  • Industry Terms
    • Not Available to Industry
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:

    pAH-CTX1-rhadCas9 was a gift from Silvia Cardona (Addgene plasmid # 129391 ; http://n2t.net/addgene:129391 ; RRID:Addgene_129391)
  • For your References section:

    A broad-host-range CRISPRi toolkit for silencing gene expression in Burkholderia. Hogan AM, Rahman ASMZ, Lightly TJ, Cardona ST. ACS Synth Biol. 2019 Sep 6. doi: 10.1021/acssynbio.9b00232. 10.1021/acssynbio.9b00232 PubMed 31491085