pCM0-TB288
$30.00
From Nick Jacobebbinghaus and Dr. Thomas Baier, Bielefeld University, Germany, April 2024
Host strain: DH5a
Selection in E. coli: Streptomycin (50 μg/ml)
Position L0 (Patron et al.): B5
Brick name: pCM0-TB288
Brick type: Antibiotic Resistance gene (CDS)
Brick function: A synthetic construct, which allows bicistronic co-expression from a joint mRNA encoding an upstream-located GOI and the downstream-located selection marker aphVIII. A variable ORF 1 (B3 and B4 position) will be terminated by addition of a GS-Linker (8 aa long), the RBCS intron 2 and a C-terminal FLAG-tag including a Stop-codon. A 249 bp long, native intergenic sequence from the C. reinhardtii nuclear genome (gene locus Cre01.g022450/Cre01.g022462) allows transcriptional read through and ribosome assembly for individual translation of downstream aphVIII. A suitable terminator including a 3’UTR is required (in B6_C1 position) and terminates transcription. This construct transfers the aphVIII selection pressure on a joint mRNA for GOI expression, allows bicistronic expression designs and simplifies screening procedure after nuclear transformations due to increased expression level of the upstream-located GOI. Plasmid size: 3681 bp, see Jacobebbinghaus et al. 2024 for detailed description and application guidelines
Source of Brick: Jacobebbinghaus et al., 2024 https://doi.org/10.1111/tpj.16677
Acceptor plasmid: pAGM1301
Designer / Builder: N. Jacobebbinghaus and T. Baier