TABLE 3.
Primers used for the fusion of signal peptides, library creation, and sequencing
| Primer | Sequence 5′–3′a |
|---|---|
| RMLN | CCTCTATACTTTAACGTCAAGG |
| RMLC | GGGAGGGCGTGAATGTAAGC |
| OB1_95F | GAGAGACTGAAGCTGAATTCGCAGCGACTGTCGTTTATGA |
| OB1_95R | TCATAAACGACAGTCGCTGCGAATTCAGCTTCAGTCTCTC |
| OB1_98F | GAGAGACTGAAGCTGAATTCGCAGCAACCGTTGTGTATGA |
| OB1_98R | TCATACACAACGGTTGCTGCGAATTCAGCTTCAGTCTCTC |
| OB1_100F | GAGAGACTGAAGCTGAATTCGCGATAGGACCTGTGGCAGA |
| OB1_100R | TCTGCCACAGGTCCTATCGCGAATTCAGCTTCAGTCTCTC |
| 3PO_95F | GAGGGGCTGAAGCTGAATTCGCAGCGACTGTCGTTTATGA |
| 3PO_95R | TCATAAACGACAGTCGCTGCGAATTCAGCTTCAGCCCCTC |
| 3PO_98F | GAGGGGCTGAAGCTGAATTCGCAGCAACCGTTGTGTATGA |
| 3PO_98R | TCATACACAACGGTTGCTGCGAATTCAGCTTCAGCCCCTC |
| 3PO_100F | GAGGGGCTGAAGCTGAATTCGCGATAGGACCTGTGGCAGA |
| 3PO_100R | TCTGCCACAGGTCCTATCGCGAATTCAGCTTCAGCCCCTC |
| 163A-F | GCTCCCCAGTTGGCCGCTNDTCCGCCCGTCCCAGATAGCAC |
| 163B-F | GCTCCCCAGTTGGCCGCTVGHCCGCCCGTCCCAGATAGCAC |
| 163C-F | GCTCCCCAGTTGGCCGCTTGGCCGCCCGTCCCAGATAGCAC |
| 163A-R | GTGCTATCTGGGACGGGCGGAHNAGCGGCCAACTGGGGAGC |
| 163B-R | GTGCTATCTGGGACGGGCGGDCBAGCGGCCAACTGGGGAGC |
| 163C-R | GTGCTATCTGGGACGGGCGGCCAAGCGGCCAACTGGGGAGC |
| 265A-F | GATAAGAGCGAATCCAAATNDTGGAACGACAGGATTCGCA |
| 265B-F | GATAAGAGCGAATCCAAATVGHGGAACGACAGGATTCGCA |
| 265C-F | GATAAGAGCGAATCCAAATTGGGGAACGACAGGATTCGCA |
| 265A-R | TGCGAATCCTGTCGTTCCAHNATTTGGATTCGCTCTTATC |
| 265B-R | TGCGAATCCTGTCGTTCCDCBATTTGGATTCGCTCTTATC |
| 265C-R | TGCGAATCCTGTCGTTCCCCAATTTGGATTCGCTCTTATC |
| 165A-F | CCAGTTGGCCGCTGCGCCGCCCNDTCCAGATAGCACCCTA |
| 165B-F | CCAGTTGGCCGCTGCGCCGCCCVGHCCAGATAGCACCCTA |
| 165C-F | CCAGTTGGCCGCTGCGCCGCCCTGGCCAGATAGCACCCTA |
| 165A-R | TAGGGTGCTATCTGGAHNGGGCGGCGCAGCGGCCAACTGG |
| 165B-R | TAGGGTGCTATCTGGDCBGGGCGGCGCAGCGGCCAACTGG |
| 165C-R | TAGGGTGCTATCTGGCCAGGGCGGCGCAGCGGCCAACTGG |
| 163165wtRev | GTGCTATCTGGGACGGGCGGCGCAGCGGCCAACTGGGGAGC |
| 163165mutRev | GTGCTATCTGGACGGGGCGGACGAGCGGCCAACTGGGGAGC |
| 163165wtFor | GCTCCCCAGTTGGCCGCTGCGCCGCCCGTCCCAGATAGCAC |
| 163165mutFor | GCTCCCCAGTTGGCCGCTCGTCCGCCCCGTCCAGATAGCAC |
| 265Rev | TGCGAATCCTGTCGTTCCRAWATTTGGATTCGCTCTTATC |
| 265For | GATAAGAGCGAATCCAAATWTYGGAACGACAGGATTCGCA |
| FS | ACGACTTCCAGGTCCCTGACCAAGC |
| RS | TCAATGTCCGCGTTCGCAGGGA |
| Lac100Forward | CGTTAGCCGTGATAAACGTAGAGCAGGGAAA |
| Lac100Foward2 | GATTCTTTCAACACGGGACAAATTGGGCTGACG |
| Lac100Mut | GCGATAGGACCTGTGGCAGACCTTCACATC |
| Lac100HF | GATGTGAAGGTCTGCCACAGGTCCTATCGC |
a
Overhangs for pJRoC30, for cloning, are underlined. Overhangs for laccase genes, for cloning, are shown in italics and underlined. Codons including targeted positions for saturation or recombination are shown in bold. Mixed bases are represented under the following code: W = A/T; Y = C/T; K = G/T; R = A/G.