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Table 2 Oligonucleotides employed in this study

From: Expansion of the genetic toolkit for metabolic engineering of Clostridium pasteurianum: chromosomal gene disruption of the endogenous CpaAI restriction enzyme

Oligonucleotide

Sequence (5′-3′)

ltrB.NheI.S

CTAGC GCTATATGCGTTGATGCAATTTCTATGCACTCGTAGTAGTCTGAGAAGGCATATG

ltrB.BstAPI.AS

ATGC CTTCTCAGACTACTACGAGTGCATAGAAATTGCATCAACGCATATAGCG

ltrA.XhoI.S

TTTCTACTCGAG GCGTTGATGCAATTTCTATGCACTC

ltrA.BstAPI.AS

GGCATCAGAGCAGATTGTACTGAG

del-Pptb.S

GGG GTTAATCATTTAACATAGATAATTAAATAGTAAAAGGGAGTGTCGAGATATCC

del-Pptb.AS

TCGAG GATATCTCGACACTCCCTTTTACTATTTAATTATCTATGTTAAATGATTAACCCC

MfeI/BstAPI.S

AATTG ATTTAGTAATTTCTATAAGCAGGTTAGCTGTAAAACTAGCAGTAGCACGCATATG

MfeI/BstAPI.AS

ATGC GTGCTACTGCTAGTTTTACAGCTAACCTGCTTATAGAAATTACTAAATC

NheI/MfeI.S

CTAGC ATTTAGTAATTTCTATAAGCAGGTTAGCTGTAAAACTAGCAGTAGCACC

NheI/MfeI.AS

AATTG GTGCTACTGCTAGTTTTACAGCTAACCTGCTTATAGAAATTACTAAATG

NheI/SacII.S

CTAGC ATTTAGTAATTTCTATAAGCAGGTTAGCTGTAAAACTAGCAGTAGCACCCGC

NheI/SacII.AS

GG GTGCTACTGCTAGTTTTACAGCTAACCTGCTTATAGAAATTACTAAATG

frag1.BglII.S

GGGATATGATATACGAGTAAGGAGATCT GG

frag1.EcoO109I.AS

AGTATTAGGCCCT GACGTCCCACATAATTCACAACATTTAGC

frag2.BglII.S

AACAGGAGATCT GCTAAATGTTGTGAATTATGTGGGACGTC

frag2.EcoO109I.AS

TACTCTAGGCCCT GGAGACCCCACACTACCATCG

frag3.BglII.S

TCGCCAAGATCT CGATGGTAGTGTGGGGTCTCC

frag3.EcoO109I.AS

GTGCCACCTGACGTCTAAGAAACC

3′SOE.S

TGGGAAATGGCAATGATAGCGAAAC

SOE.EcoO109I.AS

ATAGGCGTATCACGAGGCCCT TTC

gBlock.BglII.S

CGAGTAAGGAGATCT GGAACGATAAAACG

CpaAII-anneal.S

CTAGA GTCGACGTCACGCGTCCAAGGAGATCTCCAGGCCTGCAGACATGCA

CpaAII-anneal.AS

AGCTT GCATGTCTGCAGGCCTGGAGATCTCCTTGGACGCGTGACGTCGACT

SYCP.gBlock.S

GGAGGTCAATCTATGAAAATGCGATTAAGC

SYCP.gBlock.AS

CTTTCGTTTCGTTCCCATAGGTTCTCC

MTLCP.REN-HindIII.S

GTATTTAAAGCTT ATAATTATCCTTAAATTTCTTAAAAGTGCGCC

REN.Fw

CTACTTGAGGTCTAGGACTTCTATCT

REN.Rv

ACAGATAGGCCATTAAAGGTATTCA

ltrB.Fw

CCAACGCGTCGCCACGTAATAAAT

ltrB.Rv

ATGGGAACGAAACGAAAGCGATGC

  1. Italics: relevant restriction endonuclease recognition sequences.