The article considers the eigenvalue problem of the differentiation operator, when the spectral param- eter is also present in the boundary condition with an integral perturbation, where the integrand has the property of limited variation and has a value of unity at the ends of the segment [-1, 1]. The derivatives with respect to the time variable included in the boundary condition naturally arise when solving (by the Fourier method) initial boundary value problems for evolution equations. The conjugate operator is constructed. It is shown that the spectral questions of the conjugate operator have a similar structure. The characteristic determinant of the original direct spectral problem with an integral perturbation of the boundary condition and for the eigenvalue problem of a loaded first-order differential equation on a segment with a periodic boundary condition, which is an entire analytical function of the spectral parameter, is constructed. Based on the formula of the characteristic determinant, conclusions are drawn about the asymptotic behavior of the spectrum of the original «perturbed» differentiation operator and the loaded first-order differential equation on the segment. A special feature of the operator under consideration is the non-self-adjointness of the operator in L2(–1,1). The quadratic proximity of the systems of eigen functions of the «unperturbed» and «perturbed» differentiation operators and the Riesz basis property of these systems are proved. In this case, the system of eigenfunctions of the «perturbed» operator is not orthonormal.