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Table 4 Primers used in this study

From: Establishment of markerless gene deletion tools in thermophilic Bacillus smithii and construction of multiple mutant strains

BG nr

Sequence 5′–3′

Purpose

3464

AACTCTCCGTCGCTATTGTAACCA

Check plasmid presence

3465

TATGCGTGCAACGGAAGTGAC

Check plasmid presence

3633

GCCGTCGACCATTTGCAGTAGGTCTCGATC

ldhL-us-Fw

3636

GCCGAATTCTAGGTCACCAAAGACGAAATTG

ldhL-ds-Rv

3637

GCTCCCTTTGTATGGTCGTTTACATAATAAGAAACTCCTTTCGTCATTTC

ldhL-us-Rv

3638

GAAATGACGAAAGGAGTTTCTTATTATGTAAACGACCATACAAAGGGAGC

ldhL-ds-Fw

3664

AGGGCTCGCCTTTGGGAAG

Int. check, in plasmid

3663

ATCGCGTGAAATGTTCTAATGG

Int. check ldhL on chr.-Fw

3669

AACCGATGCCGTTGATTAAAG

Int. check ldhL on chr.-Rv

3887

GCCGAGCTCTTGCCGGAATTCTTTCAC

pta-lacZ-Fw

3888

GCCTCATGACTATTTTTCAATTACCTGCAAAATTTTC

pta-lacZ-Rv

3971

GCCGAATTCAGCTAATCTTGTTGACGGTTTTC

sigF-ds-Fw

3972

GTAACTAAGGAGTCGTGCCTTAACGATTCATGTGCTTTTTTTTG

sigF-ds-Rv

3973

CAAAAAAAAGCACATGAATCGTTAAGGCACGACTCCTTAGTTAC

sigF-us-Fw

3974

GCCGTCGACCTCTGATTTAGAAGATGGAGGTTTT

sigF-us-Rv

3990

CGCCTATTCTTTTCGCTAAAATCGG

Int. check sigF on chr.-Fw

3991

ATAAGCTGCAGAGGGATATACAC

Int. check sigF on chr.-Rv

4534

GCCTCTAGAATTGGTCATTTGATTAGA

ET 138 ldhL + prom.-Fw

4535

GCCAAGCTTTTAAGAAAGTACTTTATT

ET 138 ldhL + prom.-Rv

4522

GCCGAATTCGAGGTACATAGCCCGGAATC

pdhA-us-Fw

4523

GTCATTTGCGGCATGGCTTACATTCGTGTCACCTCTTCCTTTC

pdhA-us-Rv

4524

GAAAGGAAGAGGTGACACGAATGTAAGCCATGCCGCAAATGAC

pdhA-ds-Fw

4525

GCCGTCGACCATCCTCATAACGGCCATCC

pdhA-ds-Rv

4563

GTTTCACATACCATTTAACGATTT

Int. check pdhc on chr.-Rv

4564

GTCAATAGGTGCAAATGGATTTTC

Int. check pdhc on chr.-Fw

  1. us upstream flanking region, ds downstream flanking region, Fw forward primer, Rv reverse primer, Int. integration, chr. chromosome, prom. promoter.