pK4
$30.00
From Pablo Leivar, IQS School of Engineering-Barcelona Spain, December 2024
Host strain: DH5α
Selection in E. coli: Kanamycin (50 µg/ml)
BsaI sites presence for homology arms cloning: Yes
BbsI sites for Level 2 cloning: Yes: TGCC and GGGA.
LacZα gene flanked by BbsI sites: Yes
Chloroplast 5′ and 3′ homology arms: None. Customized by user by cloning at BsaI sites.
Function: Golden Gate compatible destination vector in a two step reaction: (1) user-chosen Chlamydomonas plastome 5’ and 3’ homology regions (arms) are introduced using BsaI enzyme. (2) Obtained vector is used as a Level 2 Golden Gate compatible destination vector to introduce modules using BbsI enzyme. For more information, refer to Melero-Cobo et al.
Melero-Cobo X, Gallemí M, Carnicer M, Monte E, Planas A, Leivar P. MoCloro: an extension of the Chlamydomonas reinhardtii modular cloning toolkit for microalgal chloroplast engineering. Physiol Plant. 2025 Jan-Feb;177(1):e70088. doi: 10.1111/ppl.70088. PMID: 39871102; PMCID: PMC11772913.