Sequence For Primer

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)