pEC3006 (pSpy1C-PZn_ffluc)
(Plasmid #218515)

• Purpose: replicative E. coli-S. pyogenes shuttle plasmid for zinc-inducible expression of the Firefly luciferase in S. pyogenes
• Depositing Lab: Emmanuelle Charpentier
• Publication: Lautenschlager et al Front Bioeng Biotechnol. 2024 Jun 7;12:1395659. doi: 10.3389/fbioe.2024.1395659. eCollection 2024.

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

Resource Information

• Supplemental Documents:
  • pSpy1C-PZn_ffluc.gb

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Luciferase Limited Use Label License
• Industry Terms:
  • Not Available to Industry

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