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3A) was assembled, using the one-step SLIC method 60 , from five PCR fragments generated from suitable plasmid templates with 1525 bp mutual overlaps, thus introducing the following changes compared with the preceding pSB15 vector: (i) replacement of the constitutive pro S promoter for the engineered PylRS gene by the inducible ara BAD promoter, including the ara C repressor gene, (ii) replacement of the tet p/o for the POI-encoding region, here sfGFPa39, by the lac UV5 p/o , including the lac I repressor gene, and (iii) removal of the tet R repressor cistron downstream of the bla ( amp r ) gene and replacement by the coding region for chloramphenicol acetyltransferase ( cam r ) further harboring an amber stop codon at amino acid position 112

The primary outcomes were the incidence of macrovascular diseases defined as the composite indicator of the first occurrence of HF, MACE, or cerebrovascular disease

Comparable alterations were noted in the aortic endothelium of db/db mice [107]

doi: 10.1158/15417786.MCR-170451 187 PrabhulkarSLiC-Z