pAED2
(Plasmid #234599)

• Purpose: Fluorescent reporter of the SOS response
• Depositing Lab: Tanel Tenson
• Publication: Kaldalu et al NPJ Antimicrob Resist. 2025 Apr 11;3(1):25. doi: 10.1038/s44259-025-00097-0.

Backbone

• Vector backbone: pSC101
• Backbone size w/o insert (bp): 3968
• Total vector size (bp): 6022
• Vector type: Bacterial Expression ; reporter

Growth in Bacteria

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

Gene/Insert 1

• Gene/Insert name: mScarlet-I
• Species: Synthetic
• Insert Size (bp): 696
• Mutation: The single amino acid substitution T74I found in mScarlet-I results in a marked maturation acceleration in cells, but at the cost of a moderate decrease in fluorescence quantum yield (0.54) and fluorescence lifetime (3.1 ns), although both values are still higher than those of all previously engineered bright mRFPs.
• GenBank ID: APD76536.1
• Promoter: cda

Cloning Information for Gene/Insert 1

• Cloning method: Ligation Independent Cloning
• 5′ sequencing primer: ACATGGTCCTTCTTGAGTTTGTAACAGC
• 3′ sequencing primer: TTAGGCGGTCACTTGTACAGCTCG

Gene/Insert 2

• Gene/Insert name: gfp-mut2
• Species: Aequorea victoria
• Insert Size (bp): 726
• Mutation: GFPmut2 was derived from avGFP with the following mutations: S65A/V68L/S72A
• Promoter: Ptet+dnaK P1

Cloning Information for Gene/Insert 2

• Cloning method: Ligation Independent Cloning
• 5′ sequencing primer: CTCATGTTTGACAGCTTATCATCGATAAGC
• 3′ sequencing primer: GGCGGTTATTTGTACAATTCATCCATACC

Resource Information

• A portion of this plasmid was derived from a plasmid made by: cda promoter was amplified from pANO1::cda’ that was gift from Dr. Lars Hestbjerg Hansen and is published in Norman, A., Hansen, L. H. & Sørensen, S. J. Construction of a ColD cda promoter-based SOS-green fluorescent protein whole-cell biosensor with higher sensitivity toward genotoxic compounds than constructs based on recA, umuDC, or sulA 1135 promoters. Appl. Environ. Microbiol. 71, 2338–2346 (2005).

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Please visit https://doi.org/10.1101/2024.11.06.622235 for bioRxiv preprint.