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A cysteine-less variant was created for the E1 component, onto which cysteines were substituted at selected loop positions.
Exonization was defined as an event in which a transcript variant was created with insertion of a TE in the intronic sequence of a gene.
The PMA1 URA3 variant was created from the pma1Δ::KanMX4/PMA1 strain; the ura3Δ0 mutation was converted to URA3 via PCR amplification of URA3 from pRS306 and transformation.
In this study, a cPYP variant was created (designated c-E-helix-PYP) in which a heterodimeric coiled-coil-forming sequence (E-helix) [ IAALEKE 2IAALE] is inserted at the newly created surface loop.
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The C-terminally truncated cited3 variant is created by placing a stop codon after the first 199th amino acids.
Cells expressing the CD-associated T300A variant were created using a targeting strategy that preserved all intronic sequence and changed a single base pair in exon 9 (A G) resulting a change from alanine to threonine at position 300.
A family of stabilized variants was created including a consensus-driven triple variant, A275P/N186D/V622I.
Based on sequence alignment and structural modeling, a library of enzyme variants was created by a semi-rational evolution strategy in position Thr238 and Pro242.
To obtain a thorough understanding of substrate specificity of the protease, a substrate library of 19
A library of 2400 KDPG-aldolase variants was created using ep-PCR with 2 3 base changes per gene.
A filtered list of variants was created for quality score determination similar to the dual approach of Chia et al. [ 18].
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