B(pET30acer_oFTN2)

The BL21(DE3) production strain is a lysogen of bacteriophage DE3. Due to a chromosomal copy of the T7 RNA polymerase gene under the control of the lacUV5 promoter, such strains are suitable for expressing T7 promoter-regulated genes using the lactose analog Isopropyl-β-D-thiogalactopyranoside (IPTG) as an inducer. doi:10.3389/fmicb.2014.00172.

The pET30acer_oFTN2 vector enables T7 RNA polymerase-based expression of FTN2, which is a humanized Fab that neutralizes tumor necrosis factor α. For FTN2 translocation into the periplasmic space, the light chain (LC) and the heavy chain (HC) of the Fab were fused to the outer membrane protein A signal sequence (ompA_SS), which utilizes the post-translational SecB-pathway. doi: 10.1002/BIOT.201800637.

Transcription termination is initialized by the strong terminator tZenit, consisting of 3 single termination loops. More precisely, tZenit includes a synthetically improved T7 terminator (T7T UUCG), a rrnBT1 termination sequence, and a native T7 terminator. doi:10.1021/sb5000115.

The cer element provides additional stability by preventing the formation of plasmid di- and multimers via recombination events catalysed by host cell enzymes. doi: 10.1002/j.1460-2075.1988.tb02884.x.

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