TABLE 3.

Oligonucleotides used in this study

OligonucleotideSequenceaCleavage site
B1-escC5′-AAAAAGCAGGCTGCTGCTATTTGTTTGACAGTGGCA-3′
B2-escC5′-AGAAAGCTGGGTAACGTTACGTTTTTATCAAGAATCG-3′
R1-escC5′-GAAGATCTTTACATTACACAATTCGTCCTA-3′BglII
R2-escC5′-GAAGATCTTGAATGATGATTTCTCTGGCGA-3′BglII
B1-escD5′-AAAAAGCAGGCTTCATGTTATCCTCATATAAAATTA-3′
B2-escD5′-AGAAAGCTGGGTTTAATACGACAGTGGAATATGTA-3′
R1-escD5′-CCGCTCGAGTGGAGTTTAATCCGTGGTTG-3′XhoI
R2-escD5′-CCGCTCGAGCCGCAATTAATGATGTCTTAACG-3′XhoI
B1-escJ5′-AAAAAGCAGGCTCATATCCCGTTAGCATCA-3′
B2-escJ5′-AGAAAGCTGGGTTGGCAAAGATACCACTTC-3′
R1-escJ5′-GAAGATCTTATTGCGCGAACTTAATCCTC-3′BglII
R2-escJ5′-GAAGATCTAATACCCGGTGTTATCGATTG-3′BglII
B1-espAcom5′-AAAAAGCAGGCTATTATAAGGAGGATGTTATGTG-3′
B2-espAcom5′-AGAAAGCTGGGTCACAGACTGGATATCGTT-3′
B1-escCcom5′-AAAAAGCAGGCTGATGCCGCCAACACACT-3′
B2-escCcom5′-AGAAAGCTGGGTGGTCATTGAGATCTTGTTGTGTC-3′
B1-escDcom5′-AAAAAGCAGGCTCATTAGCCATTGGAAACTCAC-3′
B2-escDcom5′-AGAAAGCTGGGTGTATCCTGGTAATCAAGTCTCT-3′
B1-escJcom5′-AAAAAGCAGGCTCATATCCCGTTAGCATCA-3′
B2-escJcom5′-AGAAAGCTGGGTTGGCAAAGARACCACTTC-3′
5GST-escC5′-CGGGATCCGCCCCGTCCTCTCTGGAA-3′BamHI
3GST-escC5′-CGGAATTCTTATTCGCTAGATGCAGATTTTAT-3′EcoRI
5GST-escJ5′-CGGGATCCTGATAAGAGCAATTGTATACAG-3′BamHI
3GST-escJ5′-CGGAATTCTTACCCGTCCTGTCCTGAGGAT-3′EcoRI
5GST-escD5′-AAAAAGCAGGCTTCATGTTATCCTCATATAAAATA-3′
3GST-escD5′-AGAAAGCTGGGTTTAATACGACAGTGGAATATGTA-3′
5GST-escF5′-AAAAAGCAGGCTTCCTGGAAGTTCTGTTCCAGGGGCCCATGAATTTATC TGAAATTAC-3′PreScission protease
3GST-escF5′-AGAAAGCTGGGTTTAAAAACTACGGTTAGAAATG-3′
R1V5HIS5′-GCGAGCTCACAAGTTTGTACAAAAAAGCTGAA-3′SacI
R2V535′-GCTCTAGATTAACCGGTACGCGTAGAATCGAG-3′XbaI
B1-escD-V55′-AAAAAGCAGGCTCCATATTTTTTCTCATTGTTCAACC-3′
B2-escD-V55′-AGAAAGCTGGGTTATACGACAGTGGAATATGTATAGA-3′
B1-escF-V55′-AAAAAGCAGGCTGTTCGTAATGAACGTAAATAGTTAA-3′
B2-escF-V55′-AGAAAGCTGGGTTAAAACTACGGTTAGAAATGGTTGA-3′
  • a Underlining indicates restriction enzyme sites or, for 5GST-escF, a region encoding a recognition site for PreScission protease.