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A series of fission yeast targeting vectors that can be used for wild-type strains having no selectable markers have been designed.
A PCR fragment carrying the modification but no selectable marker replaces the counterselection cassette in a second round of GET Recombination.
This is because (i) there are no selectable configurations after filtering and (i i), the alternative filter (TR,max, true) defined for the adaptation strategy keeps selecting configuration gray!75 blackcmyk 0,0,0,0.4➍.
There is no selectable difficulty level, or set amount of laps, but the boost is immediately available.
However, because no selectable markers are involved, this kind of genetic alteration is only applicable to selectable phenotypes.
Plasmid pDE00.0201 carries the iaaM, ipt, GUS (β-glucuronidase), and nptII genes, while pGR-Pv010 carries Pv010 and GFP but no selectable marker gene.
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They will no longer be selectable.
Based on the backbone of vector pCAMBIA (CAMBIA, Canberra, ACT, Australia), we designed and constructed a set of three T-DNA destination vectors for various tasks, carrying no plant selectable marker gene or genes encoding resistance to hygromycin (hptII, hygromycin phosphotransferase) and phosphinothricin (bar, phosphoinothricin acetyltransferase).
The nos::ntpII plant selectable marker gene, which confers resistance to kanamycin in transgenic plants, was also introduced in the T-DNA.
The top-down in vitro SCRaMbLE system is a convenient way to generate combinatorial diversity of DNA constructs with no need for selectable markers.
In our experiments, lines with extremely high numbers of T-DNA inserts (4 to 8) were frequently obtained when using Nos or tCUP1 promoters for selectable marker gene expression.
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