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pSK9065
(Plasmid #84345)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 84345 Standard format: Plasmid sent in bacteria as agar stab 1 $85

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    constructed by Firth lab
  • Modifications to backbone
    See supplemental information from associated article for full details.
  • Vector type
    Bacterial Expression
  • Promoter Pxyl/tet
  • Selectable markers
    Neomycin (select with G418)
  • Tag / Fusion Protein
    • mRFPmars (C terminal on backbone)

Growth in Bacteria

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

Cloning Information

  • Cloning method Restriction Enzyme

Resource Information

Terms and Licenses

Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

In S. aureus, pSK9065 replicates via the origin of replication from the staphylococcal multiresistance plasmid pSK1, whereas in pSK9067, the compatible origin from the conjugative S. aureus multiresistance plasmid pSK41 is used. These low-copy-number theta-replicating staphylococcal origins (5 to 10 copies per cell) were employed to minimize the structural instability that can arise in recombinant plasmids that utilize rolling circle replication. pSK9065 contains the aadD gene, which encodes neomycin resistance in S. aureus. pSK9065 contains the Pxyl/tet which has been modified to enable tight regulation of the expression of rfp gene fusions. The tetR promoter contains an optimized −10 box. These vectors facilitate tightly controlled expression of fusion proteins in S. aureus to enable visualization in the presence of natively expressed wild-type counterparts.
The excitation and emission spectra for mRFPmars is 585 and 602 nm, respectively.

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:

    pSK9065 was a gift from Neville Firth (Addgene plasmid # 84345 ; http://n2t.net/addgene:84345 ; RRID:Addgene_84345)
  • For your References section:

    Two-plasmid vector system for independently controlled expression of green and red fluorescent fusion proteins in Staphylococcus aureus. Brzoska AJ, Firth N. Appl Environ Microbiol. 2013 May;79(9):3133-6. doi: 10.1128/AEM.00144-13. Epub 2013 Mar 1. 10.1128/AEM.00144-13 PubMed 23455338