Table 2 Primers and source of sequence
From: Development of butanol-tolerant Bacillus subtilis strain GRSW2-B1 as a potential bioproduction host
Region description | Primer | Primer sequence (5' → 3')a | Source of sequence or reference |
|---|---|---|---|
Terminator -LacZ | Z-F | CTCTGATGCCGCATAGTTAA | pQF50, Laboratory stock |
Z-R | ctag AGATCT(BglII)CATAATGGATTTCCTTACGC | ||
P43 promoter | P43-F | GCAG GCATGC(SphI)ACTGACAAACATCACCCTCT | B. subtilis 168 chromosome |
P43-R | aTgc AAGCTT(HindIII)TGGTACCGCTATCACTTTAT | ||
PKm promoter | PKm-F | gcag GCATGC(SphI)GCTATGACCATGATTACGAA | pUC4K, Laboratory stock |
PKm-R | aTgc AAGCTT(HindIII)TGTATTACTGTTTATGTAAGCAGAC | ||
P2N promoter | P2N-F | GCAG GCATGC(SphI)TCACTTCGTACATAATGGAC | pNCMO2, Takara Bio Inc, Japan |
P2N-R | ATGC AAGCTT(HindIII)TTCGCAGGAAAGCCATG | ||
P2L promoter | P2L-F | GCAG GCATGC(SphI)GATCAGCTTGAAATATGTACATAG | B. subtilis 168 chromosome |
P2L-R | ATGC AAGCTT(HindIII)TGATAAATTTATTTATTTAGGATCCGATCT | ||
PTet promoter | PTet-F | gcag GCATGC(SphI)GTTCAACAAACGGGCCATAT | pHY300PLK, Takara Bio Inc, Japan |
PTet-R | aTgc AAGCTT(HindIII)AATAATGAGGGCAGACGTAG | ||
Pspac promoter | Pspac-F | GCAG GCATGC(SphI)CGCACCCTGAAGAAGATTTA | pDG148, Laboratory stock |
Pspac-R | ATGC AAGCTT(HindIII)AATTGTTATCCGCTCA | ||
PxylA promoter | Pxyl-F | gcag GCATGC(SphI)ATCCACCGAACTAAGTTGGT | pWH1520, Mo Bi Tec, Germany |
Pxyl-R | ATcc TCTAGA(XbaI)TTGATTTAAGTGAACAAGTTTATCCATC | ||
pHY300PLK ΔtetL | TZ-F | ATCGTTAAGGGATCAACTTTGGGAG | pHY300PLK, Takara Bio Inc, Japan |
TZ-R | ATTTCACCCTCCAATAATGAGGGC | ||
Kmr (kan) | K-F | ATTGGAGGGTGAAATATGAGAATAGTGAATGGACCAA | pDG148, Laboratory stock |
K-R | TGATCCCTTAACGATTCAAAATGGTATGCGTTTTGAC | ||
16s rRNA | 63-F | CAGGCCTAACACATGCAAGTC | (Marchesi et al. 1998) |
1387-R | GGGCGGWGTGTACAAGGC |