Table 2 The primer sequences used for constructing the pUGK4 shuttle vector
Gene name | Primer sequence (5’ − 3’) | Restriction enzyme sites |
|---|---|---|
pUC19 | Forward: ATAGAGCTCTGGCGTAATCATG GTCATAGC Reverse: ATAGAGCTCAATTCACTGGCCGT CGTTTTAC | SacI SacI |
pGK02 | Forward: ATAGAGCTCGATCAACGGTAAA TCCGTTGGC Reverse: ATTGAGCTCGGTACCATATCCTG ACATTCTCTTTAC | SacI SacI, KpnI |
Erythromycin resistance gene (ErmR) | Forward: AATGGTACCGGATCCTTTCGCAG TAACTCTATTATCAAC Reverse: ATGGTACCGTCGACTGCAGGATC CTTATCTATTAAATAATTTATAG | KpnI, BamHI KpnI, SalI, PstI, BamHI |
LDH1 promoter | Forward: ATAGTCGACCATTTGCACGAATT CTAATC Reverse: AATCTGCAGATACTTCCTTCCAT ATTAGTG | SalI PstI |
Signal peptide (SP) | Forward: ATACTGCAGATGAAGAAAAATT TAAGAATCGTTAGCGCTG Reverse: ATAGCATGCGGGCCCTGATGAAC TTGCGTTAATA | PstI SphI, ApaI |
Multi cloning site (MCS) | Forward: ATAGGGCCCGATATCGCCGGC Reverse: ATAACGCGTTCTAGACACGTGT CGCGA | ApaI MluI |
Cell wall anchor(CWA) | Forward: AATGCATGCACGCGTAGGCCTA AGTCAGCTACTTTGC Reverse: ATAAAGCTTGGTACCAGGCCTTT AGTGGTGGTGGTGGT | SphI, MluI, StuI HindIII, KpnI, StuI |
Super folder green fluorescent protein (sfGFP) | Forward: ATACTGCAGGGCCCATGTCAAA GGGTGAAG Reverse: AATCTGCAGCCGGCCTTGTATAA TTCATCCATACC | PstI, ApaI PstI, NaeI |