Lactic Acid Bacteria (LAB) are a heterogeneous group of Gram-positive bacteria with low G+C content. Lactobacillus is the largest genus, with over 150 species and can be divided into several groups based on genetic heterogeneity. Within the Lactobacillus plantarum group the diversity of phenotypical traits and preferred habitats still varies largely. L. plantarum CD033 was isolated from a stable silage in Austria. High transformation efficiencies (up to 109 cfu/g DNA) make this strain highly attractive for strain improvement by genetic engineering e.g. for the expression of recombinant proteins. This study deals with the development of genetic traits for L. plantarum. We have established a variety of suitable shuttle vectors, mainly based on high copy plasmids pCDLbu1 isolated from L. buchneri CD034. We tested endogenous constitutive promoters (e.g. Ptuf33 and Ptuf34) in either high or low copy number vectors. The optimal spacer sequence between Shine-Dalgarno sequence and translation start of reporter gene mCherry was determined. L. plantarum CD033 embodies a 7.9 kb plasmid pCD033, which was sequenced and annotated. Relative plasmid copy number of pCD033 was determined as well as the minimal stable replicon. Furthermore, we suggest replication by a novel theta mechanism. By providing a curing plasmid, and addition of sub-lethal doses of novobiocin, L. plantarum CD033 was cured of pCD033. A novel, plasmid free strain L. plantarum 3NSH was achieved, and evaluated for expression of recombinant proteins. Inducible promoters further contribute to a comprehensive use of L. plantarum as production host. Therefore, we established and characterized three different promoter/repressor systems (namely PlacA, PxylA and PlacISynth) regarding mCherry expression. We determined the preferred inducer (lactose, xylose and IPTG) and their required concentrations. Fluorescence signals and growth behavior were monitored with the BioLector® micro-fermentation system. The mCherry expression was inducible, but weak in comparison to a constitutive promoter and basal expression without inducer was observable. Furthermore, we successfully applied the T7 RNA polymerase for the first time in L. plantarum under control of an inducible promoter. This study reveals a novel plasmid free L. plantarum strain and appropriate tools for biotechnological applications e.g. enhanced silage processes or recombinant protein production in a Generally Regarded as Safe (GRAS) microorganism.