pHC01
(Plasmid #106900)

• Purpose: Ratiometric gas biosensor plasmid for 3-oxo-C12-HSL sensing
• Depositing Lab: Jonathan Silberg
• Publication: Cheng et al ACS Synth Biol. 2018 Jan 24. doi: 10.1021/acssynbio.7b00405.

Backbone

• Vector backbone: ColE1 origin with Kan resistance marker
• Backbone size w/o insert (bp): 3000
• Total vector size (bp): 6977
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): XL1 Blue
• Copy number: High Copy

Gene/Insert 1

• Gene/Insert name: LasR
• Alt name: Biobrick BBa_C0179
• Species: Pseudomonas aeruginosa
• Insert Size (bp): 720

Gene/Insert 2

• Gene/Insert name: Methyl Halide Transferase
• Alt name: MHT
• Species: Batis maritima (sequence was E. coli codon optimized)
• Insert Size (bp): 693

Gene/Insert 3

• Gene/Insert name: Ethylene Forming Enzyme
• Alt name: EFE
• Species: Pseudomonas syringae (sequence was E. coli codon optimized)
• Insert Size (bp): 1053

Gene/Insert 4

• Gene/Insert name: Red Fluorescent Protein
• Alt name: RFP
• Species: Synthetic
• Insert Size (bp): 678

Resource Information

• Supplemental Documents:
  • phc01-ratiometric-gas-reporting plasmid.gb
• Addgene Notes:
  • Addgene Diagnostic Digest

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry

Depositor Comments

LasR contains V83A, G120D mutations and MHT contains an an A157V mutation compared to the NCBI reference sequences. Plasmid functions as described in the associated publication.