Journal: Microbiology Spectrum

Article Title: Versatile allelic replacement and self-excising integrative vectors for plasmid genome mutation and complementation

doi: 10.1128/spectrum.03387-23

Figure Lengend Snippet: Strains and vectors used in this study

Article Snippet: The zeocin resistance marker ( zeoR ) was amplified from pCRBlunt II-TOPO vector (Invitrogen, Waltham, MA USA) using zeo1 and zeo2 primers and inserted into the Not I- Bam HI-restricted pKOV vector (Addgene plasmid #25769, a gift from George Church) in the same orientation as in chloramphenicol resistance gene ( cmR ). zeo1 was designed by adding the Not I restriction site at the 5′end, and similarly, Sac I, Xho I, and Bam HI restriction sites were incorporated to the 5′end of zeo2 for cloning purposes. lacZ alpha amplified from V308 pDNR MCS lacZ alpha-Donor (Addgene plasmid #11691, a gift from Tony Pawson) using lac1 and lac2 primers was inserted at the Xho I site, and the final vector was designated as pKOV-ZeO (Fig. S1).

Techniques: Clone Assay, Plasmid Preparation, Marker, Expressing

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Cell Factory Design and Culture Process Optimization for Dehydroshikimate Biosynthesis in Escherichia coli

doi: 10.3389/fbioe.2019.00241

Figure Lengend Snippet: Plasmids and strains used in the study.

Article Snippet: For disruption and substitution of chromosomal genes, suicide vector pKOV (Addgene, USA) was used, which has the sacB gene for providing a markerless system.

Techniques: Cloning, Plasmid Preparation, Expressing, Disruption, Modification