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pRH2502
(Plasmid #84379)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 84379 Plasmid sent as bacteria in agar stab 1 $65

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pTC-0X-1L
  • Backbone manufacturer
    Addgene 20315
  • Backbone size w/o insert (bp) 4640
  • Total vector size (bp) 8779
  • Modifications to backbone
    The PacI site near the T4g32 sites was removed. PacI sites were introduced 15 bp after the NdeI site encompassing the start codon of lacZI, and 5’ of the SV40 PA terminator. lacZI was removed and dcas9 was inserted under the control of uv15tetO promoter at NdeI and PacI cloning sites
  • Vector type
    Bacterial Expression, CRISPR

Growth in Bacteria

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

Gene/Insert

  • Gene/Insert name
    dcas9
  • Alt name
    cas9 D10A and H840A
  • Species
    Streptococcus pyogenes
  • Insert Size (bp)
    4139
  • Mutation
    Aspartate at position 10 changed to Alanine, Histidine at position 840 changed to Alanine
  • GenBank ID
    AAK33936.1
  • Promoter uv15tetO

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site NdeI (not destroyed)
  • 3′ cloning site PacI (not destroyed)
  • 5′ sequencing primer N/A
  • 3′ sequencing primer N/A
  • (Common Sequencing Primers)

Resource Information

  • Terms and Licenses
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:

    pRH2502 was a gift from Robert Husson (Addgene plasmid # 84379)
  • For your References section:

    Investigating essential gene function in Mycobacterium tuberculosis using an efficient CRISPR interference system. Singh AK, Carette X, Potluri LP, Sharp JD, Xu R, Prisic S, Husson RN. Nucleic Acids Res. 2016 Jul 12. pii: gkw625. 10.1093/nar/gkw625 PubMed 27407107