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Addgene

pEC3105 (pSpy1C-PxylS2_ffluc)
(Plasmid #218512)

Ordering

This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 218512 Standard format: Plasmid sent in bacteria as agar stab 1 $85

Backbone

  • Vector backbone
    pSpy1C
  • Total vector size (bp) 5706
  • Modifications to backbone
    The Plac_mrfp cassette was replaced with the synthetic PxylS2 promoter and the ffluc reporter from pLZ12Km2-P23R:TA:ffluc. See associated publication for detailed construction information.
  • Vector type
    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, also use 20 µg/mL chloramphenicol in THY medium.
  • 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 PxylS2

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 constitutive synthetic PxylS2 promoter from S. pyogenes driving expresses the Firefly luciferase. Luminescence can be detected by adding luciferin to the culture. See associated publication for more details.

Resource Information: The PxylS2 promoter was obtained via gene synthesis. Its sequence was derived from the following publication:
Xie Z, Qi F, Merritt J2013.Development of a Tunable Wide-Range Gene Induction System Useful for the Study of Streptococcal Toxin-Antitoxin Systems. Appl Environ Microbiol79:.https://doi.org/10.1128/AEM.02320-13
The ffluc reporter gene was taken from pLZ12Km2-P23R:TA:ffluc (Addgene 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:

    pEC3105 (pSpy1C-PxylS2_ffluc) was a gift from Emmanuelle Charpentier (Addgene plasmid # 218512 ; http://n2t.net/addgene:218512 ; RRID:Addgene_218512)
  • 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