From Gary Yates, Luke Mackinder lab, University of York-UK, July 2020

Recombineering plasmid containing the λ phage cassette under an L-arabinose inducible promoter and the temperature-sensitive SC101 origin of replication. Should be grown at 30°C, λ phage cassette expression activated at 37°C.

Host strain: DH5α
Selection in E. coli: tetracycline

pRed (a.k.a. pLM072) was a gift to Luke Mackinder from Mihail Sarov (Max Planck Institute, Dresden, Germany), see the following publication for details: https://www.nature.com/articles/nmeth933

Sequence


Wang J, Sarov M, Rientjes J, Fu J, Hollak H, Kranz H, Xie W, Stewart AF, Zhang Y. An improved recombineering approach by adding RecA to lambda Red recombination. Mol Biotechnol. 2006 Jan;32(1):43-53. doi: 10.1385/mb:32:1:043. PMID: 16382181.

From Gary Yates, Luke Mackinder lab, University of York-UK, July 2020

Recombineering plasmid designed for generating C-terminal mNeonGreen-3xFLAG fusion proteins.

Host strain: DB3.1 (streptomycin resistant, ccdB resistant)
Selection in E. coli: kanamycin
Selection in C. reinhardtii: paromomycin


https://www.biorxiv.org/content/10.1101/2020.05.06.080416v1

From Gary Yates, Luke Mackinder lab, University of York-UK, July 2020

Recombineering plasmid designed for generating C-terminal CrVenusYFP-3xFLAG fusion proteins.

Host strain: DB3.1 (streptomycin resistant, ccdB resistant)
Selection in E. coli: kanamycin
Selection in C. reinhardtii: hygromycin

Sequence


https://www.biorxiv.org/content/10.1101/2020.05.06.080416v1

From Gary Yates, Luke Mackinder lab, University of York-UK, July 2020

Recombineering plasmid designed for generating C-terminal mScarlet-i-3xFLAG fusion proteins.

Host strain: DB3.1 (streptomycin resistant, ccdB resistant)
Selection in E. coli: kanamycin
Selection in C. reinhardtii: hygromycin

Sequence


https://www.biorxiv.org/content/10.1101/2020.05.06.080416v1

From Gary Yates, Luke Mackinder lab, University of York-UK, July 2020

Recombineering plasmid designed for generating C-terminal mTurquoise2-3xHA fusion proteins.

Host strain: DB3.1 (streptomycin resistant, ccdB resistant)
Selection in E. coli: kanamycin
Selection in C. reinhardtii: zeocin

Sequence


https://www.biorxiv.org/content/10.1101/2020.05.06.080416v1