The fluorescent labeling of viruses is an important tool in the study of host-pathogen relationships. This is particularly true for studies of vaccinia virus, a close relative of smallpox; used as a vaccine for its eradication. Actin plays an important role in the life cycle of vaccinia virus, promoting virus motility and infection by the creation of virus-induced actin tails, as well as a role in virus-induced infected host cell motility. We report here a novel method that greatly reduces the number of steps required in the process of creating fluorescent tag-carrying recombinant viruses, which we have subsequently employed to create a recombinant virus capable of labeling various cellular structures, including the actin cytoskeleton, upon infection. This technique combines the ability of successful homologous recombination occurring using relatively short regions of homology with the principle of transient dominant selection (TDS). This technique provides a relatively rapid, robust and reliable method of fluorescently labeling viral proteins of interest or different cellular structures in a modular, sequential manner. Thus we have produced recombinant vaccinia viruses that are both fluorescent themselves and also fluorescently label polymerised actin, enabling us to visualise actin-dependent viral and/or host cellular motility in real-time. F11 is a vaccinia viral protein that inhibits RhoA signaling and promotes virus-induced host cell motility. We aim to examine its protein homolog in ectromelia virus (mousepox) by creating a fluorescent virus deletion mutant, capable of labeling the actin cytoskeleton, which can be used to monitor infection of its natural host.