Plasmid General Structure

 

Overall organization of SEVA plasmids structure. SEVA vectors are formed by three variable modules: a cargo (blue), a replication origin (green) and an antibiotic marker (magenta). Enzymes used to change the functional modules are shown with the same colour code. Modules are separated by three fixed regions (shared by all vectors) including the T0 and T1 transcriptional terminators and the oriT origin for broad host range transfer to a suite on Gram-negative recipients mediated by the RP4 conjugation machinery. Note the numbering position +1 of the DNA sequence is the first T of the unique PacI site.

Default SEVA cargo

 

fig-seva-cargo-structure

The structure of the default SEVA cargo. Cargos are cloned as PacI/SpeI fragments and contain the typical pUC18 polylinker enzymes from EcoRI to HindIII (The completely ordered restriction enzyme list is highlighted in the figure). Additional enzymes (i.e. SfiI, AvrII and NotI) are placed outside of the polylinker for specific cloning purposes. Within the cargp sequence, the enzyme recognition sites and the hybridization site of the universal M13 (R24/F24) primers are shown.