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Table 1 Primers used in this work

From: Efficient 9α-hydroxy-4-androstene-3,17-dione production by engineered Bacillus subtilis co-expressing Mycobacterium neoaurum 3-ketosteroid 9α-hydroxylase and B. subtilis glucose 1-dehydrogenase with NADH regeneration

Primers

Sequences (5′–3′)

Restriction sites

KshA-F

ATCCATATGACTACCGAGACAGCCG

NdeI

KshA-R

CGACGCGTTTAGTGGTGGTGGTGGTGGTGGCTCGGCTGCGCGGAC

MluI

KshB-F

CGCGGATCCATGACGGAGGAGCCGCTC

BamHI

KshB-R1

CGCGGATCCCTATTCGTCGTAGGTGACTTC

BamHI

HpaII-F1

ACCGGAATTCATGACGGAGGAGCCGCTC

EcoRI

KshB-R2

CCCCCCGGGTTAGTGGTGGTGGTGGTGGTGTTCGTCGTAGGTGACTTC

SmaI

HpaII-F2

ACCGGGTACCATGACGGAGGAGCCGCTC

KpnI

Gdh-R

ACCGAAGCTTTTAACCGCGGCCTGCCTGGAATGAAG

HindIII

  1. The restriction enzyme sites were bold type