Table 2 Primers for DNA cloning in this study
Primers | Sequence (5’-3’)a |
|---|---|
Fragments amplified | |
Porin-Promoter F | CAGTTACTCATGGGTTCC |
Porin-Promoter R | ATGTAGGTTCCTTAACTAGC |
phaA-F | TAAGCCGCGCTCGCTGTTGGGCAAAG |
phaA-R | ATTTACAGCTAACAGCGGCGAGCTAGTC |
phaB-F | CGCTGTTAGCTGTAAATGCTGAGGCG |
phaB-R | CACAAGTACTTCTCCCCC |
phaC-F | TTCCGACAGCTAAACGCAACTATGTTC |
phaC-R | CAGCGAGCGCGGCTTAGTCCAAGTTTAG |
OriC-F | TATTGGTTCACTGATTGAGTAAG |
OriC-R | CGTTTAGCTGTCGGAATTAATCTGACTTTTTTG |
Overlap extension PCR | |
PorinP + phaA F | GATAGCGTTGTTATTGTAAGC |
PorinP + phaA R | TGATTGATCCCATTAAAGTAG |
PorinP + phaB F | ACTGGTGGAACTGGTGGA |
PorinP + phaB R | ATCCCATTAAAGTAGTTAGC |
OriC + PorinP F | TTTATTTCTGGTGACCAT |
OriC + PorinP R | TCGGAATTAATCTGACTT |
Gibson assembly | |
OriC + P F | CGAGGGGGGGCCCGGTACTGTCGGAATTAATCTGACTTTTTTG |
OriC + P R | CATCGTAAACTGTTACCCCCAACCGAG |
phaC F | GGTAACAGTTTACGATGCGGGAAGCTC |
phaC R | GATCAATCAATGATGTGCCTGGGCAGA |
phaB + P F | GCACATCATTGATTGATCCCATTAAAGTAGTTAG |
phaB + P R | GATCAATCAATGCAACTGGATAGCGTTG |
phaA + P F | CAGTTGCATTGATTGATCCCATTAAAGTAGTTAGCG |
phaA + P R | GAACAAAAGCTGGGTACATGGCCAATCAAGCCCCC |