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pEC3006 (pSpy1C-PZn_ffluc)
(Plasmid #218515)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 218515 Standard format: Plasmid sent in bacteria as agar stab 1 $85
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Backbone

  • Vector backbone
    pSpy1C
  • Total vector size (bp) 5568
  • Modifications to backbone
    The mrfp cassette was replaced with the zinc-inducible promoter PZn (PczcD) from S. pyogenes and the ffluc reporter from pLZ12Km2-P23R:TA:ffluc.
  • Vector type
    Bacterial Expression, Luciferase

Growth in Bacteria

  • Bacterial Resistance(s)
    Chloramphenicol, 25 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Growth instructions
    For E. coli, use 20 µg/mL chloramphenicol in LB medium. For S. pyogenes, use 20 µg/mL chloramphenicol in THY medium. The zinc-inducible promoter is inducible with 80 µM ZnSO4.
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    ffluc
  • Species
    Photinus pyralis
  • Insert Size (bp)
    1653
  • Mutation
    deletion of Plac_mrfp cassette
  • GenBank ID
    XM_031473197
  • Promoter PZn (PczcD)

Cloning Information

  • Cloning method Gibson Cloning
  • 5′ sequencing primer GTAAAACGACGGCCAGTC
  • 3′ sequencing primer CAGGAAACAGCTATGAC
    • (Common Sequencing Primers)

Resource Information

Terms and Licenses

Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

This high-copy replicative shuttle plasmid features the zinc-inducible promoter PZn driving expression of the Firefly luciferase reporter in the presence of zinc. Luminescence can be detected by adding luciferin to the culture. See associated publication for more details.

Resource Information: The zinc-responsive promoter PZn (PczcD) was taken from the S. pyogenes SF370 genome. The ffluc reporter gene was taken from the original pLZ12Km2-P23R:TA:ffluc plasmid (Plasmid #88900).

Please download the detailed plasmid map using the file linked below

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:

    pEC3006 (pSpy1C-PZn_ffluc) was a gift from Emmanuelle Charpentier (Addgene plasmid # 218515 ; http://n2t.net/addgene:218515 ; RRID:Addgene_218515)

  • For your References section:

    Expanding the genetic toolbox for the obligate human pathogen Streptococcus pyogenes. Lautenschlager N, Schmidt K, Schiffer C, Wulff TF, Hahnke K, Finstermeier K, Mansour M, Elsholz AKW, Charpentier E. Front Bioeng Biotechnol. 2024 Jun 7;12:1395659. doi: 10.3389/fbioe.2024.1395659. eCollection 2024. 10.3389/fbioe.2024.1395659 PubMed 38911550
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