Table 2.
Primers used in this study.
| Oligonucleotide name | Sequence | Strand targeted | Region targeted relative to arr start |
|---|---|---|---|
| CRISPRi Oligonucleotides | |||
| arr1_Forward | GGGACGGAACGCCGATTGTGCACC | Coding strand | −47 bp to −28 bp upstream |
| arr1_Reverse | AAACGGTGCACAATCGGCGTTCCG | ||
| arr2_Forward | GGGACCCCCTGTTCCGTGTGTGAT | Coding strand | −27 bp to −8 bp upstream |
| arr2_Reverse | AAACATCACACACGGAACAGGGGG | ||
| arr3_Forward | GGGATGCACTTCGAACGGTTTCGG | Coding strand | +13 bp to +32 bp downstream |
| arr3_Reverse | AAACCCGAAACCGTTCGAAGTGCA | ||
| Oligo 2_Forward | GGGACCGGGTGCACAAT | Non-coding | −50 bp to −38 bp upstream |
| Oligo 2_Reverse | AAACATTGTGCACCCGG | ||
| Oligo 3_Forward | GGGATTCCGATCACACA | Non-coding | −32 bp to −20 bp upstream |
| Oligo 3_Reverse | AAACTGTGTGATCGGAA | ||
| Oligo 4_Forward | GGGAGAGCTCAAGGTGG | Non-coding | +66 bp to +78 bp downstream |
| Oligo 4_Reverse | AAACCCACCTTGAGCTC | ||
| Oligo 5_Forward | GGGATGGGGCTTGCGGCAAGGCC | Non-coding | −74 bp to −56 bp upstream |
| Oligo 5_Reverse | AAACGGCCTTGCCGCAAGCCCCA | ||
| Oligo 6_Forward | GGGACCGGTGCCGGGTGCACAAT | Non-coding | −56 bp to −38 bp upstream |
| Oligo 6_Reverse | AAACATTGTGCACCCGGCACCGG | ||
| Oligo 7_Forward | GGGAAAGGCCGAGCTCAAGGTGG | Non-coding | +60 bp to +78 bp downstream |
| Oligo 7_Reverse | AAACCCACCTTGAGCTCGGCCTT | ||
| Oligo 8_Forward | GGGAAGTTCGGCACCCCACACCG | Coding | +166 bp to +184 bp downstream |
| Oligo 8_Reverse | AAACCGGTGTGGGGTGCCGAACT | ||
| Oligo 9_Forward | GGGACCGATTGTGCACCCGGCAC | Coding | −53 bp to −35 bp upstream |
| Oligo 9_Reverse | AAACGTGCCGGGTGCACAATCGG | ||
| Oligo 10_Forward | GGGACCCCTGTTCCGTGTGTGAT | Coding | −27 bp to −9 bp upstream |
| Oligo 10_Reverse | AAACATCACACACGGAACAGGGG | ||
| RT-qPCR primers | |||
| sigA_forward | CCTGGAACTCGACGACCTC | ||
| sigA_reverse | GACTCTTCCTCGTCCCACAC | ||
| arr_forward | AGATCACCCAGACGTTGGAC | ||
| arr_reverse | CTCGGGCTCGACGATGTAAA | ||
These are underlined in the table first four bases in each of the CRISPRi primer sets are used for cloning the sgRNA into pRH2521. The region targeted does not include the 3 bp PAM site.