TABLE 3.
Primers used in this work
| Primer | Sequence (5′-3′)a | pDB101 coordinates |
|---|---|---|
| RepSF | CAGAATTTAAATCTGTCAAAGC | 1113-1134 |
| RepH-SR | CCCAgGCcTTGCAAAGAATGG | 1649-1629 |
| RepH-SF | CCATTCTTTGCAAgGCcTGGG | 1629-1649 |
| RepNR | GGGCGTTCTGCTAGCTTG | 2082-2065 |
| FHindIII | gggaagcttGGAAAAGGGATATATTG | 5382-5398 |
| RPstI | gggctgcagTTTTTTGTATAGTAATATTGTATC | 5549-5526 |
| FPstI | gggctgcAGAATGGGGCGTAGTTATGG | 5550-5569 |
| RSalI | GCAAACATAGTcGAcAATTTTGACTTTCC | 5696-5668 |
| FSalI | GGAAAGTCAAAATTgTCgACTATGTTTGC | 5668-5696 |
| RXbaI | CCGATCATATCTAGaCCAGGGTTAAATTG | 6170-6142 |
| FXbaI | CAATTTAACCCTGGtCTAGATATGATCGG | 6142-6170 |
| CG2 | cCggAAttCGGCATTACAGAGCATAAAGG | 6305-6332 |
| RBamHI | cccggatccCTATTCTTTTTCGTTTTC | 6420-6403 |
| FBamHI | gggggatccAATCACAAATCACAAGTG | 6421-6438 |
| RKpnI | gggggtacCTTCTTACTCTTATTTTATCATC | 6495-6473 |
| FKpnI | gggggtAccAAATAGAAAGAAGTGAGTG | 6493-6514 |
| OmegaER | ccggaATTCTCACTCCTTTTAAAGTTTGTC | 6740-6716 |
| CG1 | GATTTACACGTTTTGCAAGCT | 6915-6895 |
| #3 | CCAGTCCAATAATCGTCG | 18977-18994 |
| #4 | GCTAGTTCCAAGTTTCTCCCCC | 19194-19173 |
a
Introduced mutations are shown as lowercase letters, and restriction sites are underlined.