Emerging deep learning-based fault diagnosis methods have advanced in the current industrial scenarios of various working conditions. However, the prerequisite of obtaining target data in advance limits the application of these models to practical engineering scenarios. To address the challenge of fault diagnosis under unseen working conditions, a domain generation framework for unseen conditions fault diagnosis is proposed, which consists of an Adaptive Feature Fusion Domain Generation Network (AFFN) and a Mix-up Augmentation Method (MAM) for both the data and domain spaces. AFFN is utilized to fuse domain-invariant and domain-specific representations to improve the model’s generalization performance. MAM enhances the model’s exploration ability for unseen domain boundaries. The diagnostic framework with AFFN and MAM can effectively learn more discriminative features from multiple source domains to perform different generalization tasks for unseen working loads and machines. The feasibility of the proposed unseen conditions diagnostic framework is validated on the SDUST and PU datasets and achieved peak diagnostic accuracies of 94.15% and 93.27%, respectively.