pA-Opto_N449S
(Plasmid #253256)

• Purpose: Opto-CheY N449S expressed using the proA promoter and T7 RBS variant GAAGGAGgT
• Depositing Lab: Aravinthan DT Samuel
• Publication: Vrabioiu et al Proc Natl Acad Sci U S A. 2026 Mar 10;123(10):e2516278123. doi: 10.1073/pnas.2516278123. Epub 2026 Mar 3.

Backbone

• Vector backbone: pEB2
• Backbone manufacturer: J. H. Davis, A. J. Rubin, R. T. Sauer, Design, construction and characterization of a set of insulated bacterial promoters.
• Backbone size w/o insert (bp): 3499
• Total vector size (bp): 4593
• Modifications to backbone: E. Balleza, J. M. Kim, P. Cluzel, Systematic characterization of maturation time of fluorescent proteins in living cells. Nat Methods 15, 47-51 (2018). A.M. Vrabioiu, B.G. Hosu, A.D.T. Samuel, The dynamic response of the bacterial flagellar motor to its direct intracellular input. PNAS in press.
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): NEB Stable
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: Opto-CheY with N449S mutation
• Alt name: CheY-cpAsLOV2-FliMp_N449S
• Species: Synthetic; E. coli/Avena Sativa
• Insert Size (bp): 924
• GenBank ID: 946393 AF033096.2
• Tag / Fusion Protein:
  • T7 RBS variant GAAGGAGgT

Terms and Licenses

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

Depositor Comments

Please visit https://doi.org/10.1101/2025.10.28.684865 for bioRxiv preprint.