Kanamycin-Resistant Clone Selection: Isolating transformants carrying kanamycin-resistance plasmids on LB agar with 25 µg/mL kanamycin.

Plasmid Maintenance: Sustaining selection pressure to maintain kanamycin-marked plasmids in Escherichia coli and other Gram-negative hosts.

sacB Counter-Selection: Screening double-crossover allelic exchange events using 7% sucrose-mediated lethality in sacB-bearing intermediates.

CRISPR/Cas Genome Editing Workflows: Selecting kanamycin-edited clones and counter-selecting sacB intermediates during two-step recombineering.

Conjugation/Mating Selections: Post-conjugation selection of kanamycin-marked transconjugants and sucrose-based sacB counter-selection.

Strain Purification by Restreaking: Maintaining kanR phenotypes while isolating single colonies on LB agar.

Library Screening: Selecting kanamycin-tagged mutant or plasmid libraries with sucrose-enabled sacB counter-selection.

Verification of Allelic Exchange: Distinguishing resolved recombinants from single-crossover sacB carriers using 7% sucrose.

Workflow in Diverse Hosts: Selection and sacB counter-selection in organisms such as Escherichia coli, Pseudomonas, and Vibrio.

Quality Control of KanR Constructs: Confirming functional kanamycin resistance markers at 25 µg/mL on a standard LB base (1.0% tryptone, 0.5% yeast extract, 1.0% NaCl, 1.5% agar) with 7.0% sucrose.