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. 2017 Dec 13;7:17522. doi: 10.1038/s41598-017-17583-9

Table 2.

Oligonucleotides used in this work.

Strain or Plasmid Description
Intron retargeting*
Cdi-dnaK-722a -IBS AAAAAAGCTTATAATTATCCTTAAATTCCTTCTTAGTGCGCCCAGATAGGGTG
Cdi-dnaK-722a -EBS1d CAGATTGTACAAATGTGGTGATAACAGATAAGTCTTCTTAGCTAACTTACCTTTCTTTGT
Cdi-dnaK-722a -EBS2 TGAACGCAAGTTTCTAATTTCGATTGAATTTCGATAGAGGAAAGTGTCT
Cdi-hrcA-285s -IBS AAAAAAGCTTATAATTATCCTTACTTATCGAACAAGTGCGCCCAGATAGGGTG
Cdi-hrcA-285s -EBS1d CAGATTGTACAAATGTGGTGATAACAGATAAGTCGAACAATGTAACTTACCTTTCTTTGT
Cdi-hrcA-285s -EBS2 TGAACGCAAGTTTCTAATTTCGATTATAAGTCGATAGAGGAAAGTGTCT
Cdi-hrcA-200a -IBS AAAAAAGCTTATAATTATCCTTACTTTTCCAGATGGTGCGCCCAGATAGGGTG
Cdi-hrcA-200a -EBS1d CAGATTGTACAAATGTGGTGATAACAGATAAGTCCAGATGGATAACTTACCTTTCTTTGT
Cdi-hrcA-200a -EBS2 TGAACGCAAGTTTCTAATTTCGGTTAAAAGTCGATAGAGGAAAGTGTCT
Cdi-groEL-601s -IBS AAAAAAGCTTATAATTATCCTTATTTGTCTCTGCAGTGCGCCCAGATAGGGTG
Cdi-groEL-601s -EBS1d CAGATTGTACAAATGTGGTGATAACAGATAAGTCTCTGCATATAACTTACCTTTCTTTGT
Cdi-groEL-601s -EBS2 TGAACGCAAGTTTCTAATTTCGATTACAAATCGATAGAGGAAAGTGTCT
Cdi-groEL-689a -IBS AAAAAAGCTTATAATTATCCTTACTGGTCATAATTGTGCGCCCAGATAGGGTG
Cdi-groEL-689a -EBS1d CAGATTGTACAAATGTGGTGATAACAGATAAGTCATAATTCTTAACTTACCTTTCTTTGT
Cdi-groEL-689a -EBS2 TGAACGCAAGTTTCTAATTTCGGTTACCAGTCGATAGAGGAAAGTGTCT
EBS universal CGAAATTAGAAACTTGCGTTCAGTAAAC
ClosTron sequencing
cspfdx-seq-F1 GATGTAGATAGGATAATAGAATCCATAGAAAATATAGG
pMTL007-R1 AGGGTATCCCCAGTTAGTGTTAAGTCTTGG
Screening of clones
Cdi-dnaK-F CTACAGCTGGTAACAATAGATTAGGT
Cdi-dnaK-R CTGTAGCAGTTATGAAAGGTAAGTT
Cdi-groEL-F AGTCTCAAACTATGAATACTGAATTAGATG
Cdi-groEL-R GCTTTTTACCTTGTTGAACTATTTGT
Cdi-hrcA-F TAGGGTATTTAATTCAGCCTCATACTTC
Cdi-hrcA-R TGCTACAGTTGTATAGTTTGTTAGTTGC
ErmRAM-F ACGCGTTATATTGATAAAAATAATAATAGTGGG
ErmRAM-R ACGCGTGCGACTCATAGAATTATTTCCTCCCG

*Introns were inserted after the indicated number of bases in the sense (s) or the antisense (a) orientation from the start of the open reading frame (ORF) of the target gene. Cdi, C. difficile; IBS, intron-binding sites; EBS, exon-binding sites; ErmRAM, erythromycin retrotransposition-activated selectable marker.