Table 1: Oligonucleotides used for the secondary library (ASAC) construction for CatE-activating peptides at neutral pH.

No.Nucleotide sequence (5′→3′)Size (nucleotides)Amino acid sequence of the block

Definition a

GGCTCGCGAA TACTGCGAAG AAAGGTCGT (D5)29
GATCTCACTC CTTCGCAGTA TTCGCGAGCC (D3)30

Starting 5′/3′ -halves for ASAC library

1 D5-GACCCATCCC CA/GACCCATCCC CA-D341/42DPSP
2 D5-ATTATTATTA TT/ATTATTATTA TT-D341/42IIII
3 D5-AAAGGCCCAC CA/AAAGGCCCAC CA-D341/42KGPP
4 D5-CCATGCCCAT GC/CCATGCCCAT GC-D341/42PCPC
5 D5-CCAGGCCCAC CA/CCAGGCCCAC CA-D341/42PGPP
6 D5-TCCTGCTCCT GC/TCCTGCTCCT GC-D341/42SCSC
7 D5-GTCGGGTGTG AC/GTCGGGTGTG AC-D341/42VGCD
8 D5-TTCATGTACG TG/TTCATGTACG TG-D341/42FMYV
9 D5-TGCTGCCCAC GC/TGCTGCCCAC GC-D341/42CCPR
10 D5-CCAGGCATTA AA/CCAGGCATTA AA-D341/42PGIK
11 D5-GACCCATCCC AA/GACCCATCCC AA-D341/42DPSQ
12 D5-GGAGGACGGC GG/GGAGGACGGC GG-D341/42GGRR
13 D5-TCCGCCACTT GC/TCCGCCACTT GC-D341/42SGTC
14 D5-GCCGCCTTTA GA/GGCGGCTTTA GA-D341/42GGFR
15 D5-CTGAAGGACT AC/CTGAAGGACT AC-D341/42LKDY
16 D5-AAAGATGACG AT/AAAGATGACG AT-D341/42KDDD
17 D5-TCCCCAATTA TT/TCCCCAATTA TT-D341/42SPII
18 D5-CCACCACCAT GC/CCACCACCAT GC-D341/42PPPC
19 D5-CCACCATCCT GC/CCACCATCCT GC-D341/42PPSC
20 D5-TGTGACTTCA TG/TGTGACTTCA TG-D341/42CDFM
21 D5-CCACGCCCAG GC/CCACGCCCAG GC-D341/42PRPG
22 D5-TCCCAAATTA TT/TCCCAAATTA TT-D341/42SQII
23 D5-GGAGGCGGAG GC/GGAGGCGGAG GC-D341/42GGGG

aFor simplicity, the device sequences are defined and symbolized (D5  and D3). For the YLBS block shuffling, 5′-half and 3′-half oligonucleotides are hybridized at their stem sequences (i.e., D5 and D3) and then ligated (for detail, see [19]).