Pre-service biology teachers must apply Technological Pedagogical and Content Knowledge (TPACK) acquired at university in real classroom situations to utilize the instructional potential of digital technologies for teaching biology. So far, there is little evidence on how pre-service biology teachers translate TPACK into teaching practice. The present study addresses this gap by accompanying 42 pre-service biology teachers in planning, implementing, and reflecting on a biology lesson as part of their internship semester at school. Data were collected via written lesson plans, videotaped lesson observations, and stimulated-recall reflection interviews and evaluated by applying a sequential explanatory mixed-method design. The results indicate that pre-service biology teachers enact their TPACK by focusing on technology with the content of the subject receding into the background. In addition, pre-service biology teachers focus particularly on aspects that serve to structure the lesson, rather than on aspects of student activation. The use of emerging technologies in the classroom seems to lead to insecurity among pre-service biology teachers for various reasons, whereby surface characteristics and structuring lesson aspects are focused. Within the sample, we can distinguish between two types of TPACK enactment: the split-focus type separates between content and technology, whereas the novelty-focus type systematically links content and technology, utilizing the technology as a tool for subject teaching.