The multi-resistance presented by P. aeruginosa has greatly increased due to the presence of genes for carbapenemases such as blaKPC. The dissemination of this gene has been associated with the Tn4401, the main mobile genetic element that carries blaKPC in its structure. However, some non-Tn4401 elements (NTEKPC) associated with blaKPC have been found in different bacteria. Here, we characterized in silico and in vitro blaKPC-associated elements in P. aeruginosa. To identify these elements, a search algorithm was performed using NCBI databases; sequences were filtered and pair-aligned to describe the blaKPC genetic environment. Additionally, a PCR-based strategy was designed to target Tn4401 variants and NTEKPC groups and assessed in 61 Colombian clinical isolates. Using an in silico approach, 51 blaKPC-positive entries longer than 3kb (in the blaKPC upstream region) were found; from these, 72.7% carried an NTEKPC. On the PCR assay, Tn4401 was the most frequent element among the P. aeruginosa in Colombia. However, NTEKPC-IIf was presented on 29.5% of the isolates, in different genetic lineages and at least in four hospitals. These results show high NTEKPC prevalence in P. aeruginosa.