Protein sequence

The first A domain of onnamide (500aa)

AKQLQDLGIGPDQLVAICVT
RSLDMIVGLLGILKAGGAYVPLDPEYPTERLAYMLEDSQAEVVLTQTALRHQVTALSSG
AARRPHILAVNEVLEAWNGEA
ISKGTLRREVQNTHLAYVIYTSGSTGKPKGVMIPHNALINFLTSMAKEPGLNSNDRLLA
VTTYCFDIAGLEFYLPLIMGA
QCVICSSETIVDADRLKQEISHCRPTVMQATPSTWSMLFHCGWKNEEKVKVLCGGEALP
MALQHQFMACNCEVWNLYGPT
ETTIWSAVQRIRENEAIVIGKPIQNTGIYILDHNRALVPIGVPGELCIDGDGLARGYLHKP
ELTAEKFMDHPFHRGKKIY
HTGDLARWLPNGTIEYLGRLDLQVKIRGFRIELGEIEFQLKQLPGIRDAVVVAKDMAK
DMEGLHQLVAYFIPGGGPESVD
TEVLRAGLKQKLADYMVPAFFIPLEEIPLTPNGKVDRKTLMERDILVNRSQEHTQPQSAI
EESLLEIWQDILKIDNISPT

The second A domain (497aa)

GVGREAIVGICVERSLEMIIAVMAVLKAGGAFLPIDPILPRKRREFMLNDASVSVLLTKE
ALLGKGIDDQ
IPVLFLDQNWDSIAQKSSSNLHCDVSSDSLAYVIYTSGSTGEPKGTMIEQGSWADLYIAL
EEIYQFRSIESFLQMADFSF
DVFFLDFIRAFCSGGKLVLCPKEFLFNPEMLYGLICKEKIEFADFVPGVMSHLMDYLESHG
RNLETMKTIIVGSDVFYAK
DCHRLKKLIGEQTRLFNAYGLTETTIDNLYFETNALAITDAMIVPIGRPLPNTSVYILDQHL
HPVPIGVPGELYIGGSNV
ARGYLNLPEKTQEKFIRHPFSHADDERLCRTQDLVYYQQDGTIHYMGRADSQVKIRGH
RIEVSEIETVLLQIPQVMEAVV
LVKEREAGGRFLVGYIVPEAGSPIDSHQLRVTLKEKLPDYMIPSQHVFLDKMPLLVSGKV
DRQALPEPDFDEFRAHPYVQ
PSTKLEKLIAYIWAETLGLKRVGKYDG

Nucleotide sequence
The first A domain(1491bp)
GAATTCCAAACAGTTGCAGGACCTGGGCATCGGCCCCGATCAGCTGGTTGC
CATCTGCGTGACGCGCTCTCTGGATATGATCGTT
GGTTTGTTAGGCATTCTCAAGGCCGGTGGCGCCTATGTGCCCCTGGATCCCGAA
TACCCAACGGAACGCCTGGCCTACAT
GCTGGAAGACTCTCAGGCCGAAGTTGTGCTGACGCAGACCGCTTTGCGGCAT
CAGGTGACGGCTCTGTCGTCAGGAGCGG
CTCGCAGACCCCACATCCTGGCCGTCAATGAAGTGCTAGAGGCCTGGAACGG
CGAAGCAATCAGCAAAGGCACCTTGCGG
CGCGAGGTGCAAAACACCCATCTGGCCTATGTCATCTACACGTCCGGCAGTAC
GGGAAAGCCCAAAGGCGTGATGATCCC
CCATAACGCTTTGATCAATTTTCTCACTTCCATGGCCAAGGAACCGGGACTGA
ACAGCAATGACCGGCTGCTTGCGGTGA
CCACCTATTGTTTTGATATTGCCGGCTTAGAGTTCTATCTGCCTCTGATCATGGG
CGCCCAATGTGTCATTTGTTCAAGT
GAAACGATTGTGGACGCTGACAGGTTAAAACAGGAAATCAGCCACTGTAGGC
CAACCGTCATGCAGGCGACGCCCTCAAC
CTGGAGCATGCTCTTTCACTGCGGTTGGAAGAATGAAGAAAAGGTCAAAGTG
CTGTGTGGGGGCGAGGCGCTTCCCATGG
CGCTGCAGCATCAGTTCATGGCGTGCAACTGCGAGGTGTGGAACCTCTACGGG
CCCACCGAAACGACCATCTGGTCGGCG
GTGCAACGCATCCGGGAAAATGAAGCGATCGTGATTGGAAAACCCATCCAGA
ATACCGGTATCTACATTCTCGATCACAA
CCGTGCGCTGGTTCCCATTGGCGTTCCTGGCGAGCTCTGCATTGACGGGGACG
GACTGGCCAGAGGCTATCTGCACAAGC
CGGAACTCACCGCAGAGAAATTCATGGATCACCCCTTTCATCGAGGGAAAAA
AATATACCACACCGGGGATCTGGCCCGT
TGGCTTCCCAATGGAACCATCGAGTACCTGGGGCGGCTGGATCTTCAGGTCAA
GATTCGCGGATTCCGCATTGAGCTGGG
CGAGATTGAGTTCCAGCTCAAGCAGTTGCCGGGTATCAGGGATGCCGTAGTGG
TCGCCAAAGATATGGCCAAAGACATGG
AGGGACTCCATCAGCTTGTCGCCTACTTTATTCCCGGCGGGGGACCGGAATCG
GTGGATACGGAAGTATTACGGGCCGGT
TTGAAGCAGAAACTGGCGGACTACATGGTACCTGCGTTCTTCATTCCCCTGGA
AGAAATTCCTTTGACGCCGAATGGCAA
GGTGGATCGGAAGACCTTGATGGAACGGGACATCCTGGTGAACCGATCCCAG
GAACACACCCAACCTCAGTCTGCAATTG
AAGAGAGCCTGCTGGAAATCTGGCAGGATATTTTGAAGATCGACAATATCAG
TGAAAGCTT

Forward primer: GAA TTC CAA ACA GTT GCA GGA CC(EcoRI)(23bp, 48%GC,
55oC)
Reverse primer: AAG CTT TCA CTG ATA TTG TCG ATC TTC (HindIII)(27bp,
37%GC, 55oC)

Second A domain (1401bp)

GGATCCATCGTGGGGATTTGTGTCGAGCGTTCTCTGGAAATGATCATTGCCG
TCATGGCGGTGCTCAAG
GCTGGCGGCGCCTTCCTGCCCATTGATCCGATTTTGCCCAGGAAACGGCGGGA
ATTCATGCTCAACGACGCATCGGTCTC
GGTCTTGCTGACAAAGGAAGCACTTTTGGGAAAAGGTATCGATGATCAGATCC
CTGTTCTTTTCCTAGATCAGAATTGGG
ACAGCATTGCACAGAAAAGTAGTAGTAATCTTCATTGTGATGTGAGTTCCGAC
AGTCTGGCTTATGTCATATATACTTCC
GGTTCCACAGGTGAACCGAAAGGCACCATGATTGAGCAGGGAAGCTGGGCCG
ATTTGTATATTGCTCTGGAAGAGATCTA
CCAGTTCAGAAGCATCGAAAGTTTTCTACAGATGGCCGATTTCTCCTTTGATGT
TTTTTTCTTGGATTTTATTCGGGCTT
TTTGTTCGGGAGGCAAACTGGTTTTATGCCCAAAAGAGTTCCTCTTCAACCCG
GAAATGCTCTACGGACTAATCTGCAAA
GAAAAGATTGAATTCGCCGATTTTGTTCCCGGTGTCATGAGTCATCTGATGGAT
TACCTGGAAAGCCATGGGCGAAATCT
GGAGACCATGAAAACGATCATTGTCGGGTCGGATGTGTTTTATGCGAAGGACT
GTCACAGACTCAAGAAACTGATCGGCG
AGCAAACCAGACTGTTCAATGCCTATGGCCTGACCGAAACGACCATTGACAAC
CTCTATTTTGAAACCAACGCCCTGGCC
ATAACGGATGCGATGATCGTCCCCATTGGGCGGCCCTTGCCCAATACCAGTGTC
TATATTCTGGATCAGCATCTGCACCC
GGTTCCCATTGGGGTTCCGGGTGAGTTGTATATCGGCGGTTCCAACGTTGCCA
GAGGTTATCTCAACCTGCCTGAAAAGA
CACAGGAAAAATTCATCAGACATCCCTTTAGCCACGCCGATGACGAGAGGCTC
TGTCGAACCCAGGACCTCGTCTATTAT
CAGCAGGATGGGACGATTCACTACATGGGGCGGGCGGATTCTCAAGTGAAAAT
TCGGGGTCATCGCATTGAAGTTAGCGA
GATCGAAACCGTGCTGCTGCAAATACCACAAGTCATGGAAGCCGTGGTGCTTG
TAAAAGAACGTGAAGCCGGTGGCAGGT
TTCTGGTCGGCTACATCGTACCTGAGGCTGGTAGTCCCATCGATTCGCACCAAT
TGAGAGTGACGTTAAAAGAGAAGCTG
CCGGACTACATGATTCCATCGCAACATGTCTTCTTGGATAAGATGCCACTGCTG
GTAAGCGGCAAAGTGGATCGCCAGGC
GCTGCCCGAGCCCGATTTTGATGAATTCAGAGCACATCCCTATGTTCAGCCTT
CCACTGAGAGCTC

Forward primer: TCT AGA GAT CGT GGG GAT TTG TGT C (XbaI) (25bp, 48%,
58oC)
Reverse primer: AAG CTT TCA GTG GAA GGC TGA ACA TAG (HindIII) (27bp,
44%, 58oC)

PRIMER BASED ON ALIGMENT RESULT

A-onnI
Protein sequence
ADYSLL
SSKEERLLLSDWNDTSSAYPSDCVHELFERQAETNPAAIAVMCEGKSLTYGELDQRSSVLAKQLQDLGIGP
DQLVAICVT
RSLDMIVGLLGILKAGGAYVPLDPEYPTERLAYMLEDSQAEVVLTQTALRHQVTALSSGAARRPHILAVNE
VLEAWNGEA
ISKGTLRREVQNTHLAYVIYTSGSTGKPKGVMIPHNALINFLTSMAKEPGLNSNDRLLAVTTYCFDIAGLE
FYLPLIMGA
QCVICSSETIVDADRLKQEISHCRPTVMQATPSTWSMLFHCGWKNEEKVKVLCGGEALPMALQHQFMACNC
EVWNLYGPT
ETTIWSAVQRIRENEAIVIGKPIQNTGIYILDHNRALVPIGVPGELCIDGDGLARGYLHKPELTAEKFMDH
PFHRGKKIY
HTGDLARWLPNGTIEYLGRLDLQVKIRGFRIELGEIEFQLKQLPGIRDAVVVAKDMAKDMEGLHQLVAYFI
PGGGPESVD
TEVLRAGLKQKLADYMVPAFFIPLEEIPLTPNGKVDRKTLMERDILVNRSQEHT
QPQSAIEE
Nucleotide sequence
GAATTCGCGGACTATTCCCTGCTGTCCAGCAAGGAAGAACGCCTGTTGCTT
TCCGATTGGAATGACACCTCGTCCGCCTATCCTTCA
GACTGTGTCCATGAATTGTTTGAACGTCAGGCAGAAACCAATCCCGCGGCAAT
TGCAGTGATGTGTGAAGGCAAGTCCCT
AACCTACGGAGAGCTGGATCAGAGAAGCAGCGTGCTTGCCAAACAGTTGCAG
GACCTGGGCATCGGCCCCGATCAGCTGG
TTGCCATCTGCGTGACGCGCTCTCTGGATATGATCGTTGGTTTGTTAGGCATTCT
CAAGGCCGGTGGCGCCTATGTGCCC
CTGGATCCCGAATACCCAACGGAACGCCTGGCCTACATGCTGGAAGACTCTCA
GGCCGAAGTTGTGCTGACGCAGACCGC
TTTGCGGCATCAGGTGACGGCTCTGTCGTCAGGAGCGGCTCGCAGACCCCACA
TCCTGGCCGTCAATGAAGTGCTAGAGG
CCTGGAACGGCGAAGCAATCAGCAAAGGCACCTTGCGGCGCGAGGTGCAAA
ACACCCATCTGGCCTATGTCATCTACACG
TCCGGCAGTACGGGAAAGCCCAAAGGCGTGATGATCCCCCATAACGCTTTGAT
CAATTTTCTCACTTCCATGGCCAAGGA
ACCGGGACTGAACAGCAATGACCGGCTGCTTGCGGTGACCACCTATTGTTTTG
ATATTGCCGGCTTAGAGTTCTATCTGC
CTCTGATCATGGGCGCCCAATGTGTCATTTGTTCAAGTGAAACGATTGTGGAC
GCTGACAGGTTAAAACAGGAAATCAGC
CACTGTAGGCCAACCGTCATGCAGGCGACGCCCTCAACCTGGAGCATGCTCTT
TCACTGCGGTTGGAAGAATGAAGAAAA
GGTCAAAGTGCTGTGTGGGGGCGAGGCGCTTCCCATGGCGCTGCAGCATCAG
TTCATGGCGTGCAACTGCGAGGTGTGGA
ACCTCTACGGGCCCACCGAAACGACCATCTGGTCGGCGGTGCAACGCATCCG
GGAAAATGAAGCGATCGTGATTGGAAAA
CCCATCCAGAATACCGGTATCTACATTCTCGATCACAACCGTGCGCTGGTTCCC
ATTGGCGTTCCTGGCGAGCTCTGCAT
TGACGGGGACGGACTGGCCAGAGGCTATCTGCACAAGCCGGAACTCACCGCA
GAGAAATTCATGGATCACCCCTTTCATC
GAGGGAAAAAAATATACCACACCGGGGATCTGGCCCGTTGGCTTCCCAATGGA
ACCATCGAGTACCTGGGGCGGCTGGAT
CTTCAGGTCAAGATTCGCGGATTCCGCATTGAGCTGGGCGAGATTGAGTTCCA
GCTCAAGCAGTTGCCGGGTATCAGGGA
TGCCGTAGTGGTCGCCAAAGATATGGCCAAAGACATGGAGGGACTCCATCAGC
TTGTCGCCTACTTTATTCCCGGCGGGG
GACCGGAATCGGTGGATACGGAAGTATTACGGGCCGGTTTGAAGCAGAAACT
GGCGGACTACATGGTACCTGCGTTCTTC
ATTCCCCTGGAAGAAATTCCTTTGACGCCGAATGGCAAGGTGGATCGGAAGAC
CTTGATGGAACGGGACATCCTGGTGAA
CCGATCCCAGGAACACACCCAACCTCAGTCTGCAATTGAAGATGAAAGCTT

A-onnJ
Protein sequence
SKIAFLTQPETQLLAQWQAIPRNHGDRSCIHQLFEAQAEARPDATAVVLYQAEDRLEAQSQQVATYGELNE
KA
NQLARYLRKLGVGREAIVGICVERSLEMIIAVMAVLKAGGAFLPIDPILPRKRREFMLNDASVSVLLTKEA
LLGKGIDDQ
IPVLFLDQNWDSIAQKSSSNLHCDVSSDSLAYVIYTSGSTGEPKGTMIEQGSWADLYIALEEIYQFRSIES
FLQMADFSF
DVFFLDFIRAFCSGGKLVLCPKEFLFNPEMLYGLICKEKIEFADFVPGVMSHLMDYLESHGRNLETMKTII
VGSDVFYAK
DCHRLKKLIGEQTRLFNAYGLTETTIDNLYFETNALAITDAMIVPIGRPLPNTSVYILDQHLHPVPIGVPG
ELYIGGSNV
ARGYLNLPEKTQEKFIRHPFSHADDERLCRTQDLVYYQQDGTIHYMGRADSQVKIRGHRIEVSEIETVLLQ
IPQVMEAVV
LVKEREAGGRFLVGYIVPEAGSPIDSHQLRVTLKEKLPDYMIPSQHVFLDKMPLLVSGKVDRQALPEPDFD
EFRAHPYVQ
PSTKLE
Nucleotide sequence
TCTAGATCAAAGATCGCCTTCCTCACCCAGCCGGAGACACAGCTGTTGGCG
CAGTGGCAGGCCATTCCCCGCAACCATGGGGACCGC
AGCTGTATTCATCAGCTCTTTGAAGCACAAGCCGAAGCCAGACCGGATGCCAC
CGCGGTTGTCCTGTATCAGGCTGAAGA
CCGGCTGGAAGCGCAATCGCAGCAGGTGGCCACCTATGGGGAGCTCAATGAA
AAGGCCAATCAGCTCGCACGCTATTTGC
GGAAGCTCGGCGTAGGCCGAGAGGCGATCGTGGGGATTTGTGTCGAGCGTTC
TCTGGAAATGATCATTGCCGTCATGGCG
GTGCTCAAGGCTGGCGGCGCCTTCCTGCCCATTGATCCGATTTTGCCCAGGAA
ACGGCGGGAATTCATGCTCAACGACGC
ATCGGTCTCGGTCTTGCTGACAAAGGAAGCACTTTTGGGAAAAGGTATCGATG
ATCAGATCCCTGTTCTTTTCCTAGATC
AGAATTGGGACAGCATTGCACAGAAAAGTAGTAGTAATCTTCATTGTGATGTG
AGTTCCGACAGTCTGGCTTATGTCATA
TATACTTCCGGTTCCACAGGTGAACCGAAAGGCACCATGATTGAGCAGGGAA
GCTGGGCCGATTTGTATATTGCTCTGGA
AGAGATCTACCAGTTCAGAAGCATCGAAAGTTTTCTACAGATGGCCGATTTCT
CCTTTGATGTTTTTTTCTTGGATTTTA
TTCGGGCTTTTTGTTCGGGAGGCAAACTGGTTTTATGCCCAAAAGAGTTCCTC
TTCAACCCGGAAATGCTCTACGGACTA
ATCTGCAAAGAAAAGATTGAATTCGCCGATTTTGTTCCCGGTGTCATGAGTCAT
CTGATGGATTACCTGGAAAGCCATGG
GCGAAATCTGGAGACCATGAAAACGATCATTGTCGGGTCGGATGTGTTTTATG
CGAAGGACTGTCACAGACTCAAGAAAC
TGATCGGCGAGCAAACCAGACTGTTCAATGCCTATGGCCTGACCGAAACGACC
ATTGACAACCTCTATTTTGAAACCAAC
GCCCTGGCCATAACGGATGCGATGATCGTCCCCATTGGGCGGCCCTTGCCCAAT
ACCAGTGTCTATATTCTGGATCAGCA
TCTGCACCCGGTTCCCATTGGGGTTCCGGGTGAGTTGTATATCGGCGGTTCCAA
CGTTGCCAGAGGTTATCTCAACCTGC
CTGAAAAGACACAGGAAAAATTCATCAGACATCCCTTTAGCCACGCCGATGAC
GAGAGGCTCTGTCGAACCCAGGACCTC
GTCTATTATCAGCAGGATGGGACGATTCACTACATGGGGCGGGCGGATTCTCA
AGTGAAAATTCGGGGTCATCGCATTGA
AGTTAGCGAGATCGAAACCGTGCTGCTGCAAATACCACAAGTCATGGAAGCC
GTGGTGCTTGTAAAAGAACGTGAAGCCG
GTGGCAGGTTTCTGGTCGGCTACATCGTACCTGAGGCTGGTAGTCCCATCGATT
CGCACCAATTGAGAGTGACGTTAAAA
GAGAAGCTGCCGGACTACATGATTCCATCGCAACATGTCTTCTTGGATAAGATG
CCACTGCTGGTAAGCGGCAAAGTGGA
TCGCCAGGCGCTGCCCGAGCCCGATTTTGATGAATTCAGAGCACATCCCTATGT
TCAGCCTTCCACAAAGCTGGAGAATGAAAGCTT

Of onnI (1,658 bp)

Forward primer: GAA TTC GCG GAC TAT TCC CTG CTG (EcoRI)(24bp, 54%GC,
59oC)
Reverse primer: AAG CTT TCA TCT TCA ATT GCA GAC TGA G (HindIII)(28bp,
39%GC, 57oC)

Of onnJ for pMAL (1,694 bp)

Forward primer: TCT AGA TCA AAG ATC GCC TTC CTC AC (XbaI) (26bp, 46%,
58oC)
Reverse primer: AAG CTT TCA TTC TCC AGC TTT GTG GAA G (HindIII) (28bp,
43%, 58oC)

Of onnJ for pHis

Forward primer: GGA TCC TCA AAG ATC GCC TTC CTC AC (BamHI) (26bp, 54%,
61oC)
Reverse primer: GTC GAC TCA TTC TCC AGC TTT GTG GAA G (SalI) (28bp, 50%,
61oC)