Table 1.
Oligonucleotides used in this study
| Primer namea | Sequenceb (5′to 3′) | Coordinatesc (nt) |
|---|---|---|
| Nde-F | AAGGAGGGAAACATATGAGTTACA | 3718–3741 |
| P244-Stop-Xho | CTTGGTTAGTTGTCTCGAGTATCTAAGATTGAAC | 4485–4452 |
| 19nic4d | GAATATAAAGTATAGTGTG/TTAT | 3577–3599 |
| IR1+12pAd | ACTTTATGAATATAAAGTATAGTGTG/TTATGG(A15) | 3570–3599 |
| IR1+8 | ACTTTATGAATATAAAGTATAGTGTG | 3570–3595 |
| IR1+12 | ACTTTATGAATATAAAGTATAGTGTG/TTAT | 3570–3599 |
a
The oligonucleotide IR1+12pA contains a 15-nt-long poly(A) tail.
b
Restriction sites are in boldface, and base changes that generate restriction sites are underlined. The double-base change introduced in the P244-Stop-Xho oligonucleotide to obtain the stop codon (P244Stop) is shown in italics. The oriT cleavage site is indicated by a slash.
c
Coordinates are given with respect to the pMV158 sequence (accession number NC_010096).
d
These oligonucleotides were labeled with the Cy5 fluorophore at its 5′ end.