The crude oils from the reservoirs of Mela-01 and Mela-04 wells located in the Kohat Basin, Pakistan, were geochemically analyzed to evaluate the origin, depositional conditions, and thermal maturity of the rock units and possible facies from which these oils were sourced. Gas chromatography-mass spectrometry (GC-MS) was performed on the samples to obtain biomarker and non-biomarker parameters. Analyzed non-biomarker parameters, including carbon preference index (CPI), terrigenous to aquatic ratio (TAR), isoprenoids pristane to phytane (Pr/Ph), and biomarker parameters, including steranes and dibenzothiophene/phenanthrene (DBT/P) of aromatic compounds, were utilized in the present study to achieve the objectives. Most of these parameters suggest a mixed source of organic matter (marine/terrestrial) with sub-oxic conditions in the source rocks for the analyzed oil samples in the studied wells from Mela oilfield, Kohat Basin. Furthermore, the CPI and different biomarker parameters such as steranes C29 S/S+R, ββ/αα+ββ), moretane to hopane (M29/C30H), pentacyclic terpanes C27 (Ts/Ts+Tm), H32 (S/S+R) hopanes, and aromatic methylphenanthrene index (MPI) indicate that the analyzed oils have originated from thermally mature rocks falling in the oil window. As the studied Kohat Basin has multiple source rocks and contributes to the major petroleum production of the country, the present investigations reveal that its okthe Mela oils were generated by the strata of mixed organic matter that were deposited in marine sub-oxic conditions. Furthermore, this study suggests that this stratum would also have been produced in unexplored surrounding areas such as Tirah, Orakzai, and the Bannu Depression.