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. 2016 Feb 19;7:27–33. doi: 10.1016/j.bdq.2016.01.001

Fig. 2.

Fig. 2

Map of final linear construct inserted into BY4739 chromosome IV. BY4739a is a 50 bp region homologous to Open Biosystems strain S. cerevisiae BY4739 and corresponding to the non-coding region on chromosome IV between 544595 bp and 544644 bp. The full annotation of chromosome IV can be found at http://browse.yeastgenome.org/fgb2/gbrowse/scgenome/. URA3 is a 1106 bp sequence that encodes S. cerevisiae S288C orotidine-5′-phosphate decarboxylase, an enzyme that catalyzes the synthesis of pyrimidine ribonucleotides. The sequence was PCR amplified from a pYES2 vector (Life Technologies, part # V825-20) and includes a working copy of the 804 bp URA3 gene (Gene ID: 856692) as well as the URA3 promoter sequence. ERCC-00095 represents a 438 bp sequence from NIST SRM 2374: DNA Sequence Library for External RNA Controls. ERCC-00095 (GenBank accession number KC702204.1) is derived from donor microorganism M. jannaschii. The sequence was amplified from the ERCC-00095 plasmid using primers 95F and 95R (Table S1). BY4739b is 51 bp homologous to the non-coding region on chromosome IV between 545447 bp and 545497 bp in Open Biosystems strain S. cerevisiae BY4739.