Molecular epidemiology studies aiming at understanding the acquisition of antimicrobial resistance by clinical isolates of Klebsiella pneumoniae are regularly published; however, information on the genes that contribute to its characteristic phenotype of resistance to antibiotics (intrinsic resistome) is scarce. To fill this gap, a K. pneumoniae transposon mutant library was screened and 171 mutants presenting changes in their susceptibility to antibiotics were selected, in which the transposon insertion site was determined in 75. Twenty-seven mutants for which insertion points had been previously identified were included in the analysis. A total of 102 mutants were selected for further studies. In 70 mutants the transposon was inserted in a gene with a known function, whilst in 19 the insertion occurred in genes encoding proteins with unknown functions and 13 insertions occurred in intergenic regions. Moreover, 87 of the insertions were localised in the chromosome, with 15 insertions located in the two plasmids carried by this strain. Whereas some of the mutated genes are already known to be involved in antimicrobial resistance (ampG, acrB, tolC), several of them are involved in regular processes of bacterial physiology, including K. pneumoniae virulence. Together with results published for other organisms, these results support that determinants involved in basic processes of bacterial physiology may contribute to antimicrobial resistance. These findings also indicate that, besides acquired resistance genes, plasmids may harbour other genes belonging to their backbone that can also be involved in resistance.