TABLE 2.
Primers used for cloning, quantitative gene expression, and plasmid copy number assays
| Primer | Sequence |
|---|---|
| E.coli GAPA Fwd | 5′-TGTTAGACGCTGATTACATGG-3′ |
| E.coli GAPA Rev | 5′-CTTTAACGAACATCGGAGTGT-3′ |
| pRAS3A Fwd | 5′-GGAGCCACTATCGACTACG-3′ |
| pRAS3A Rev | 5′-GAAGCAGCCCAGTAGTAGG-3′ |
| pRAS3MobC Fwd | 5′-ACACAACAGAGCAGCTAGA-3′ |
| pRAS3MobC Rev | 5′-TCTGGTCAAGCGTGTATCC-3′ |
| pRAS3MobE Fwd | 5′-GCATCAGCGGAAGCAGCC-3′ |
| pRAS3MobE Rev | 5′-GCCTATCGCACTTCGCC-3′ |
| pRAS3ORF3 Fwd | 5′-CCGTTCGATCTGGTAGACC-3′ |
| pRAS3ORF3 Rev | 5′-GTTCTTCCATGTCTCGACG-3′ |
| pRAS3OriT Fwd | 5′-CTTGCAGGATGAGCCAGAC-3′ |
| pRAS3OriT Rev | 5′-TGGTTGCGGAGTTGACAG-3′ |
| pRAS3OriV Fwd | 5′-GTCGAATTCGTACATTATGTTTCG-3′ |
| pRAS3OriV Rev | 5′-ATAGGTACCAGTCTTTTCCATCC-3′ |
| pRAS3REPB Fwda | 5′-ACGAATTCATGTGCGGGAAG-3′ |
| pRAS3REPB Reva | 5′-TCACTGCAGTGCAACATTGTA-3′ |
| pRAS3REPB2 Fwd | 5′-GCAACTATCAGGCCATCAT-3′ |
| pRAS3REPB2 Rev | 5′-TTGGGCTTGCGGTTCTC-3′ |
| pRAS3REPC2 Reva | 5′-TATCTGCAGCTTGAACAGGTG-3′ |
| pRAS3REPC3 Fwda,b | 5′-TAGAATTCAGGAGGAGGGCTATGACTCAGCAGC-3′ |
| pRAS3SS2 Fwda | 5′-GAATTCAGTGGGAGAAGCTGGAAG-3′ |
| pRAS3SS2 Reva | 5′-GGATCCGGAATGGTGTAGATCGTT-3′ |
| R6KKANR Fwd | 5′-CCATTCTCACCGGATTCAG-3′ |
| R6KKANR Rev | 5′-TCACCGAGGCAGTTCCATA-3′ |
a
The primer includes an endonuclease restriction site (underlined).
b
The primer overlaps the start codon (shown in bold) and includes an artificial ribosome binding site (in bold italics).