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Table 1 Oligonucleotide primers used for the construction of expression plasmids

From: Production of (R)-3-hydroxybutyric acid by Arxula adeninivorans

Designation

Oligonucleotide sequence

Source

ORFs primers

Eurofins-Genomics

 Akat1_start_MunI

TAGACAATTGATGGACAGACTCAACAACGTAGC

 Akat1_stop_BamHI

GCAAGGATCCTTACTCCCGGATAAACAGAGC

 Akat2_start_BamHI

TAGAGGATCCATGGAAAGAGCATCTAATCTTGCC

 Akat2_stop_NotI

GCAAGCGGCCGCCTACTCCCTAATGAAGAGCG

 Akat4_start_EcoRI

TAGAGAATTCATGTCTGTTTATATTCTTAGTGCTAAG

 Akat4N10_start_EcoRI

TAGAGAATTCATGACACCCATTGGTTCGTTTTTGGG

 Akat4 N17_start_EcoRI

TAGAGAATTCATGGGATCTCTGTCTTCTCAGAC

 Akat4_stop_BamHI

GCAAGGATCCTCAAACTCTCTCAATTACAAGAGC

 Atpk2_start_BamHI

GCAAGGATCCATGCTACGACGGGGAATTC

 Atpk2_stop_NotI

TAGCGGCCGCTCACAACTTTGCCCCAGC

 Afox2_start_EcoRI

TAGAGAATTCATGTCGGTGCCAACGGC

 Afox2_stop_NotI

TAGCGGCCGCTTAAAGCTTGGCTCCTCCG

Two-step cloning

 TEF1_SpeI

TATAACTAGTTAGTAGCGCTAATCTATAATCAG

 PHO5_BsiWI

CGGACGTACGAGCTTGCATGCCTGCAGA

 TEF1_SpeI_BsiWI

TATACTAGTACTTCGTACGCTCGACTTCAATCTATAATCAGTC

 PHO5_SacII_MluI

GGATCCGCGGCCGAACGCGTAGCTTGCATGCCTGCAGATTTTAATC

  1. Restriction sites were designated in italic