Honeybee Cytochrome P450s from version 3 of the Apis mellifera genome

Honeybee Cytochrome P450s from version 3 of the Apis mellifera genome

Version 3 sequences added June 12, 2005. This file submitted by Reed Johnson

after comparing the new version and the older version 2 and making some small changes.

Last modified Feb. 22, 2005

These sequences were submitted Feb. 11, 2004 by May Berenbaum in collaboration with Gene Robinson (genome sequencing), Hugh Roberstson (genome annotation) and Reed Johnson (P450 annotation).

On Sept. 2, 2004 Reed Johnson submitted the revised sequences from Honeybee Version 1.2. These sequence were further edited and revised by D. Nelson on Sept 13-17.Many partial sequences have been combined and the number of complete genes increased from 20 to 33. The number of P450 genes including fragments for ver 1.2is 55, including 22 partial sequences.

On Feb. 9, 2005 Reed Johnson submitted the revised sequences from Honeybee Version 2.More sequences are complete and some contamination from cow DNA was noted (CYP2E1).The gene statistics are 44 complete genes, 3 incomplete but expected to be full length, 2 pseudogenes plus 5-6 small fragments.

There are 4 CYP clans in insects. CYP2, CYP3, CYP4 and mitochondrial

CYP2 is the clan with CYP18 in it. Sometimes it is called the CYP18 clan.

CYP2 has CYP15, 18, 303, 304, 305, 306, 307, 342, 343

CYP3 has CYP6 and 9 in it and CYP28, 308, 309, 310, 317(a CYP6 subfam)

CYP4 has CYP4, 311, 312, 313, 316, 318

the mito clan has CYP12, 49, 301, 302, 314, 315

[3] [4]

The honeybee sequences have been sorted into these main CYP clan bins.

48 genes have been named. 1 named gene is a pseudogene.

[5] [6]

CYP2 clan (8 sequences, 7 complete, 1 incomplete)

[7] [8]

>CYP18A1 Am3_13.1 (31931-34244), 524 aa, 5 exons

version 2 = CYP18A1 Am2_13.1 (31931-34244), 524 aa, 5 exons

version 1.2 = CYP18A1 Am1.2_13.1b (32196-34509) plus, 524 aa, 5 exons

version 1.1 = Am1.1_13.1a (71904-74217) plus, 524 aa, 5 exons (71904-72236,

72653-72774, 72894-74323, 73404-73668, 73794-74217)

60% to CYP18A1 D. melanogaster

MGGTRIEVLCTFLVFLGVLLVARCLQWLRYVRSLPPGPWGVPVFGYLPFLKGDVHL

RYGELAKKYGPMFSARLGTQLVVVLSDHRTIRDTFRREEFTGRPHTEFINILGGYG(1)

IINTEGAMWKDQRKFLHDKLRGFGMTYMGGGKKIMESRIM(0)

REVKTFLRGLASKRGTPTDVSASLGMSISNVICSIIMGVRFQHGDARFKRFMDLIEEGFKLFGSMAAVNFIPV

MRYLPCLQKVRNKLAENRAEMAGFFQETVDQHRATFDEGTMRDLVDAYLLEIEKAKGEGRATTLFQGKNHD(1)

RQMQQILGDLFSAGMETVKTTLEWAIILMLHHPDAAIAVQEELDQVVGKS

RMPVLEDLPFLPITEATILEVLRRSSVVPLGTTHATTR(2)

DVTLHGYTIPAGSQVVPLLHAVHMDPELWEKPEEFRPSRFLSAEGKVQKPEYFMPFGVGRRMCLGDVLARMEL

FLFFSSLMHTFELRSPQGSSLPSLRGNAGVTVTPDPFDVCLLPRNLDLIEDNDMISTGAILRNIGSH*

[9] [10]

>CYP306A1 Am3_13.1 (38910-36902), 499 aa, 7 exons

version 2 = CYP306A1 Am2_13.1 (38911-36903), 499 aa, 7 exons

version 1.2 = CYP306A1 Am1.2_13.1a (39182-37170) minus, 499 aa, 7 exons

version 1.1 = CYP306A1 Am1.1_13.1b (76876-78884) minus, 499aa, 7 exons (78884-78557,

78493-78345, 78209-78007, 77901-77595, 77538-77383, 77298-77123,

77057-76876)

44% to 306A1 Anopheles gambiae = CYP306A1 in 18clan/2clan

CYP306A1 is the probable ortholog of CYP306A1 in diptera (flies and mosquitos)

the % identity is below the usual cutoff for subfamily membership,

but it makes sense to name orthologs with the same name.

MILDHYIAIFVLPFLLLLYVVRKNRKARRLPPGPWQLPLLGYLPWIDAEKPHETLTRLSRVYGPVCGFRMGSVYTVLLSDPQLIRQSFAKDSITNRAPLYLTHGIMKGYG(1)IICAEGEQWKDQRKFISNCLRNFGMVKHEGAKRDKMEERISDAVNECVS(0)VLRDRGANGPIDPLDTLHHCLGNLVNSIVFGKTYEEEDRIWKWLRHLQEEGVKQIGVAGPLNFLPFLR(2)FLPQYGRVIRSIVDGKDKTHEIYRQILDEHRARVDSGNGCKIDSFLAAFDEQMRKKDGAESGYFTEPQLYHLLADLFGAGTDTTLTTLRWFLLFMAAHPMEQ(0)EKIQSEMDLCLREGEQPTLNDRIVMPRLEAAIAEVQRIRSVTPLGIPHGTSE(0)DVEIGGYDIPCGAMIVPMQWAIHTDPAYWRDPLEFRPDRFLSEDGTFFKPESFLPFQNG(1)KRVCVGEELARMILFLFAGRILRAFSVRVPAGEIADLEGECGITLVPKPHRLAFVGRDR*

[11] [12]

>CYP307B1 Am3_14.5 (873140-876884), 507 aa, 3 exons

version 2 = CYP307B1 Am2_14.4 (871542-875286), 507 aa, 3 exons

version 1.2 = CYP307B1 Am1.2_14.10 (757965-761727) plus, 507 aa, 3 exons

version 1.1 = Am1.1_14.9 (18918-22680) minus, 507 aa, 3 exons (22680-22278,

20154-19363, 19246-18921)

CYP307B1 55% to 307B1 Anopheles, probably the ortholog of 307B1

MIPLTATTCFLIAITFLALALILLDHLRSKKTTKSVVPGDDDQHALPEPPGPKPWPILGSLHILGRYDVPYKAFADLVRDFDCQVIKLRMGSVPCVVVNGLENIKEVLTVKGHHFDSRPNFARYHLLFGGNKENS(1)LAFCNWSDVQKARREMLRAHTFPRAFSTRFNELNGIIGDEMEFMVNHLDSLSGTSVHAKPLILHCCANIFITYLCSKNFHLEHDGFRNMVENFDKVFFEVNQGYAADFLPFLMPLHHRNMARMAHWSHEIRRFVIKNIIADRVNSWNDVVPEKDYVDCLINHVKSGTEPQMSWNTALFVMEDIIGGHTAIGNLLVKVLGFLATRPEIQRLAQDEIDALGLAGNFVGLENRRSLPYVEAIILETIRIIASPIVPHVANQDSSIAG(1)FRIKKDTFIFLNNYDLNMSTDLWTSPEEFMPDRFVQNGRLLKPEHFLPFGGGRRSCMGYKLVQYVSFAILASILKNFTITPVQKEDYTIPIGNLALPEMTYKFRFERR*

[13] [14]

>CYP303A1 Am3_Un.471 (10297-12131), 375 aa, 8 exons, unable to identify first exon(s)

version 2 = CYP303A1 Am2_Un.6309 (6408-4574), 375 aa, 8 exons

version 1.2 = CYP303A1 Am1.2_Un.1361 incomplete on N-terminus (5389-7223) minus, 378 aa, 8 exons

AADG03013520.1 (WGS)

version 1.1 = CYP303A1 Am1.1_Un.2253 incomplete on N-terminus (55-1889) plus, 378 aa, 8 exons

43% to 303A1 D.mel., 44% to 303A1 Anoph. CYP303A1 in 2 clan/18 clan

may be the ortholog of 303A1, first exon below is 75% to 304A1

(1)LLLVDGNLWNEQRRFVLKHLRDFGFGRQN(1)LYMNANEYTGNNVTQSQLGTIISMHNIFGITVLNSLWKMLAGKR(2)YNIDDKELIYFQRILSITLNEIDMLGAPFSHFPLLRFIAPEISGYKSFVKIHEELWKFFK(0)DEVNNHKNTFNSDSPGNLIDIYLTILNSENYGKTFSE(1)PQLVAICVDLFMAGSETTSKVLGFCFLYLVLFPHVQKKAHEEIDRVIGRNKLPTAEDKAK(2)MTYMNAIVLESLRMFAGRSLNLPHRVQRDTKISDYKIPK(0)NTIIITNFNGILMDESWGDPENFRPERFIDGSGNIVTPSRFLPFSAG(1)KHRCMGENLAKTNIFIIATTLLQAFTFSEIPGEKPTIEHFIDGTTISPKPYRVNVSLRI*

[15] [16]

>CYP305D1 Am3_7.2 (73129-69633), 490 aa, 8 exons, D->E fixed error

version 2 = CYP305D1 Am2_7.2 frameshift repaired in 7th exon (72431-68935), 490 aa, 8 exons

version 1.2 = CYP305D1 Am1.2GroupUn.127b (70044-73538) minus, 490 aa, 8 exons (first exon is a guess)

version 1.1 Am1.1_Un.7452 incomplete on both ends (700-954) minus, 85 aa, exon 2

version 1.1 Am1.1_Un.6110 incomplete on both ends (1-622) minus, 126 aa, exons 6,7

Two fragments from v.1.1 have been combined in this sequence. The first exon is a best guess. BI513047.1 (EST) AADG03008281.1 (WGS) 39% to 305A2, 39% to 305B1 Bombyx

39% to 305B1 silkworm

MNKNFVKIFNLLLYLFIDVFNNLFFIECDG(1)PFSWPFIGNQILLKRLSRKFGGQHKAFMELSKRYNSDIITVNISYEKIIVVSGSKFCDMILQNEEFQGRPWNEFIKVRNMGKKQG(1)ITMNDGTEWKELRNWMMRTMKIFGFGKSEMIEMIQHQLVIFSENLNKNKLHQLKLLFVPAVINVLWNFITGELVAFNQQQK(2)LEHFLDLLDRRSRCFDITGGLLAAFPWIRYIAPEISGYNIMCMLNKELKDFLM(0)KTINDHKEKYIEGKEADLIDMFIQEMRKNEKSSIFTE(1)EQLMMILIDLFLAGFTTTSTTLDFLFLIVTLFPDVQRKVQKEIDSVIPYDRLPNMEDKAK(2)LPYVEAVISETYRLWPVFPIIGPRRVLCDTNIDKYVIPKDTTILFNTYSINKDPTLYPDPDKFMPERFIKNGVFEPDEYSLQFGKG(1)KRRCPGDILAKATIFILFVGIMQKYTLLPVPGKGPHSIKINSGITLTPQPYNVLVEKR*

[17] [18]

>CYP15A1 Am3_7.2 (65875-68510), 504 aa, 8 exons, GC intron splice donor after exon 6

version 2 = CYP15A1 Am2_7.2 (65177-67812), 500 aa, 8 exons

W->L, changed splice donor on penultimate intron and both splice sites on final intron

version 1.2 = CYP15A1 Am1.2_Un.127a (66491-68918) plus, 504 aa, 8 exons XM_392687.1 partial

gnl|Amellifera1|165771382 BCM Apis mellifera 12/11/2003

Am1.1_Un.897 incomplete, missing exon 5 (51606-53773) plus, 357 aa, 5 exons

Am1.1_Un.10970 incomplete on N-terminus (112-677) plus, 121 aa, 2 exons

Two fragments from v.1.1 have been combined. 18clan/2clan

47% to CYP15A1 Diploptera punctata (probable ortholog)

39% to CYP15B1 Anopheles gambiae 36% to 303A1, 35% to 305A3, 31% to 304B1

MLYVVISLLLALYCIFCIYDCVKPHNFPPG(1)PKWLPLIGCFLTFRRLKLKHKYTYVAFQELSKTYGPILGLKLGSQKLVVISTHDLVKKVLLQDEFNGRPDGFFFRVRAFGKRKG(1)ILFTEGSMWSQCRRFTMRHLRSFGLGQSTMEKYLTVEAENLVNYLRRVSTKGPVPMHTAFDIAVLNSLWCMFAGHRFDYENEKLAEILEIVHDSFR(2)LMDTMGGIISQMPFLRFIIPELSGYNNLMEILRKLWNFLDEEINNHEKHLSGNQPQDLIEAFLLEISSRNGVQNDSIFDR(1)ENLLILCLDLFLAGSKTTTDTLSTSILFLSLHSEWIKILQEELDNVVGRSRSPTLEDYSSLPIMESFLAE(0)IQRFLILAPLGVPHKTTKDVILNGYNIPK(0)DTTVLLDFHSAHNDPAYWDHPEEFRPQRFLDANGRFCQNNANIPFGLG(1)KRRCPGEMLARTSLFLYFAYVIHYFDIEISPEHGKPDLNGHDGFTISPKSYYLKITARSDVTNCSTI*

>CYP342A1 Am3_8.3 (44357-46424), 505 aa, 7 exons

version 2 = CYP342A1 Am2_8.3 (44357-46424), 505 aa, 7 exons

version 1.2 = CYP342A1 Am1.2_8.3 incomplete on N-terminus (1269-3097) plus, 505 aa, 7 exons

version 1.1 Am1.1_Un.8493 incomplete on both ends (509-1028) minus, 141 aa, 2 exons

36% to 304A1 36% to 304B1 18clan/2clan

MISFLFIIFLLLIIYKIYNSVIHVSSNTPPC(1)LPRLPIIGSYWHLLWHDYEYPYNGIIHYVNKLQSKIVTCYFGSHKTIIANDYKSIKEVLTKQEFNGRPINVDIVLQRAFGKSLG(1)IFFTEGTLWHEQRRFALRHMRDFGFGRRHEIFETNVMEEIAILVDMLKEGPINDEEK(0)KFLKNGYACFPDILYPYVANVILNIMFGERFDRSQYHKLIYFCESSMMFQKSLDTSGGAIFQFWFLKYFGNIFGYTNAIKATYQMINFIE(0)EYIDNKKDLDDYDKGLIGRYLKILKEKNNITSTFSQKQLIMTLVDFMFPATSALPSALVHAIKLVMHHPRVVNNIQEEIDRVVGTGRLVTWSDRKN(2)LPYIEATIRESLRYETLTPLSVFHKTLKKTTLCDYDIPKDTLVVTNLVALNTDPDLWGDPENFRPERFLDENNELRKDFTFPFGFG(1)HRVCPGETYSRYNMFEVFAVLMQNFNFSFVEGEPTGLDDKESGLIVTPKKTWIQVKARNMK*

[19] [20]

>CYP343A1 Am3_Un.713 (725-7080), 496 aa, 8 exons, newly completed

version 2 = CYP343A1 3241+6423 Am2_Un.573, R->K (725-7080), 496 aa, 8 exons combined Am1.2_Un.3241 and Am1.2_6423 to complete

version 1.2 = Am1.2_Un.3241 (94-459) minus, 96 aa, 2 exons at N-terminus up to C-helix

version 1.1 = Am1.1_Un.7901 incomplete on C-terminus (1-459) minus, 126 aa, 2 exons 40% to 15B1

version 1.2 = Am1.2_Un.6423 incomplete on N-terminus (67-247) minus, 254 aa, 4 exons

AADG03019874.1 (WGS)

Am1.2_Un.1305 incomplete on N-terminus (2043-2647) plus, 146 aa, 2 exons

Am1.1_Un.960 incomplete on both ends (8311-11562) minus, 147 aa, 2 exons

One fragment in v.1.1 was divided into two in v.1.2, but they are probably from

one gene. 34% to 305a4, 36% to CYP15A1, new family in the CYP2 clan

MWFVILCFVIVLIKILFDYSRPINFPPG(1)PRGLPFIGNILDIIKLINETKYYSDTWCRLAEKYGSVVGLRLGLDQPLIIVSGKSAVTEMLNRSEFDGRPSGFLYKYRCGGMQQGILFTDTDVWHSQRR(2)FALKTLKQFGFGKNSMEHILQHDAIALTNIIIELTKDGTVKNIRSIISAAVLSNLWLLIDGTK(2)FDIGMENSNLKEAINIVQDIVKSSNVSGGIINQFPFLRHLFPNLTGFSAFVERQKRINNFFM(0)EVIAKHKWKKINEEGTNFIDVYLQEIQKKNSSHSFFNE(1)NQLLYIIKDLFSAGVDTTNSTIGFIIAFLVVHQDVQSKVYDEISRVIDKDIYPSLSDKDR(2)LPYLKAVIAEVSRLANIGPTSIPHRAVKDSTFLGFEIKKNYTLLANFKSIHMDKEHWGDPEIFRPERFINEKGDFINDSWLMPFGLG(1)RRKCLGETLAKNTVFLFVACMLQRLHFMLPSNHPPPCLQGIDGFVIAPPMMDIIAVQRF*

[21] [22]

note: intact seq 36% to 15A1, 35% to 15B1, 34% to 305A4

[23] [24]

CYP3 clan (31 seqs, 28 complete and 3 incomplete, 2 of the incomplete are pseudogenes, 5-6 fragments)

[25] [26]

>CYP6AQ1 Am3_12.17 (1072541-1075366), 514 aa, 5 exons

version 2 = CYP6AQ1 Am2_12.16 (1067523-1070348), 514 aa, 5 exons

version 1.2 = CYP6AQ1 Am1.2_12.14 (417204-420026) plus, 514 aa, 5 exons

version 1.1 = CYP6AQ1 Am1.1_12.14 (315628-318450) plus, 514 aa, 5 exons (315628-316108,

316238-316604, 316677-316861, 316989-317277, 318272-318450)

45% to 6K1, 42% to 6g2m 43% to 6G1ps new subfamily in CYP6

cyan = missing seq. from EST BE844578

yellow = EST BE844462, underlined seq = EST BE844394, green = EST BE844353

magenta = EST BE844352, gray = EST BE844331 all ESTs from antennae

[27] [28]

MNLLTPYWSLDILIVSSSLMIAVYLYASWKLKYWSRRGIMQITPSPLFGNFKKCILFQKSVSEIIRELYGQNEGLPFMGFY

IFYKPFFLVRDIELVKHILVKDFNTFANKHTSADSKNDRIGYSNLFIIKNPAWKYLRGKLTSVFTSGKLKKMFDLMLIIG (1)

KNLEKHLELLNLDG

NGKEVELKDLCANFTTDLIGTTAFGVNLNSLKDPNSDFRENGRLVFDYNLKRAFEFFSIFFFPNLS

KYVSIKFFGKATDYFRNSFWSVINQRIESNVKRNDLIDCLIELREKHKNDESFEGFR (1)

FDGDDLVSQAAIFFTGGFETSSTTISFTLYELALNKDIQKTVRTEIHEALAQTDGKITYDM (0)

ITNLPYLDMVVSETLRKYPPLGFLDRVALHDYKIPNSDVTIDKDTPVIIPMIAFHYD

PKYFPNPEKYDPLRFSEEVKKTRPSYVYMPFGEGPHICIG (1)

MRLGLLQSKLGIIEILKDYEVSPCEKTKIPMVLDPKGLTTTALGGLYLNIRKITIAAG*

[29] [30]

>CYP6AR1 Am3_5.8 (437540-434791), 502 aa, 5 exons

version 2 = CYP6AR1 Am2_5.8 (434944-432195), 502 aa, 5 exons

version 1.2 = CYP6AR1 Am1.2_5.5 (430078-432827) minus, 502 aa, 5 exons

version 1.1 = CYP6AR1 Am1.1_Un.19 (44801-47550) plus, 502 aa, 5 exons

50% to AmGroupUn.5496, 47% to AmGroupUn.792b, 38% to 6a13ps all best hits to 6as

MSWLMIETVGLIATVFFLLYYYSMSKLDYWRKRGVKGPKPLPFLGNFKDVLLAKESTMDCFERA

YKEFKDEPMVGMYGSHEPLLILRDLDLIKDVLIKDFNKFAQRTQGAIRE(0)

VEPLSEQLFRLDAERWRPLRLKLSSFFSSGKLKEMFHLFVECSDNFEKYLEKMVEKGGLVECRDAAAKFSTDVIGACAFSIHTNALTDENSQFRKMGKQALATNLQQFLNDRLREYPFLFKIFGRFFVDHEVTNFFANSIKDAMDY

RIQNNVHLRDVIDILADIRENPTKCGLKE(1)

ADNLFLTSQAVLFFLAGFENASLTISNALYELAWKPEIQEKARAEIVNVLQKYDGKITYDGLEEMKYLEACIFE(1)

TLRMYPVLQWLSREAMETYTFTGTKVTIPKGQQVFLPIYAIQRDPDI

YPNPDNFDPERFTDDKIKTRHSMTHLPFGDGPRHCSG(1)

IRLAKKQLKVGLVTVLSKFKVEVCEKTRKIYQKDKKPLFLLQPVDGIHLKISKVSV*

[31] [32]

>CYP6AS1 Am3_Un.8289 (3492-1105), 498 aa, 5 exons

version 2 = CYP6AS1 Am2_Un.5363 (3492-1105), 498 aa, 5 exons

version 1.2 = CYP6AS1 Am1.2_Un.6491 (264-2651) plus, 498 aa, 5 exons

version 1.1 = CYP6AS1 Am1.1_Un.5496 (264-2651) plus, 498 aa, 5 exons

44% to 6A14, 64% to AmGroupUn.792b

MDYFQILCAISIVILTIYYYYSSKYTFWKKRGISGPKPIIFFGNFVDSIIQKRSTSEAVKKWYDDYKHESVFGIFGGTTPLLVINDLDMIKDVLIRDFSLFVDRGFHIFPK(0)IEPLSEHLFLLEAERWRPMRMKLSPIFTSGKLKEMFFLIMESAGNLEKYLDEVIKKDEMVECRELAAKFMTDVIGSCAFGINTNSLLEEDSEFRRMGKKISTPNLKVMLGNICKEFFPPLYEIVGSIFTLKDVNEFFINLVSDTMKYRKDNNIIRSDFINMLMQLKEHPEKMENIE(1)LTNTLLTAQAVVFFVAGFETSSSTMAFSLYELAQNQEIQDKLREEIRNMHEKNKGVLTYTDVKEMKYLDKVFKE(1)TLRKYPILPMLFRQAMENYTFKDTKITIPKGMKLWVPVHGIHHDPNIYPEPEVFDPERFEDDAFASRHPMSYLPFGDGPRNCIG(1)ARFAHYQSKVGLITILRHHKVNVCEKTTIPFKADERSFLLTLKGGVHLKITKI*

[33] [34]

>CYP6AS2 Am3_Un.433 (26964-24619), 498 aa, 5 exons

version 2 = CYP6AS2 Am2_Un.476 (25830-23485), 498 aa, 5 exons

version 1.2 = CYP6AS2 Am1.2_Un.601a (2171-4675) plus, 498 aa, 5 exons

version 1.1 = CYP6AS2 Am1.1_Un.792a incomplete on C-terminus of exon 2, missing 3rd exon

(8375-10702) minus, 356 aa, EST BE844607 from antennae

86% to AmGroupUn.5496

MDYFQILCAISIVILTIYYYYSLKYAFWKDRGISGPKPIIFFGNFGNSIIKKKSLSETVKKWYDDYKHESVFGIFEGTIPVLVINDLDMIKDILIRDFSLFVDRGFHIFPK(0)IEPLTQHLFLLEAERWRPMRMKLSPIFTSGKLKEMFSLIVESAGNLEKYLDEVIKKNEMVECRDLAAKFTTDVIGSCAFGINTNSLLEEDSEFRRMGKKIFSPSLKLMIGNTCKVFFPSLYEVIGNIFTMKDVDEFFINLVSDTMKYRKDNDIVRSDFINMLMQLKEHPEKMDNIE(1)LTDTLLTAQAVVFFIAGFETSSSTIAFGLYELAQNQEIQDKLREEIRKMHEKNKGILTYTDIKEMKYLDKVFKE(1)TLRKYPILSTLSRKAMENYTFKGTKITIPKGTKVWVPVYGIQHDPNIYPKPEVFDPERFEDDAFASRHPMSYLPFGDGPRNCIG(1)ARFAHYQSKVGLITILRNHKVNVCEKTTIPFKADERSFLLALKGGVHLKITKI*

[35] [36]

>Am2_4631 Am3_Un.4707 (605-937), 111 aa, 1 exon, Exon 1 of 6AS2

version 2 = Am2_Un.4631 new (605-937), 111 aa, 1 exon

only one aa diff to 6AS2 probably the same seq.

MDYFQILCAISIVILTIYYYYSLKYAFWKDRGISGPKPIIFFGNLGNSIIKKKSLSETVKKWYDDYKHESVFGIFEGTIPVLVINDLDMIKDILIRDFSLFVDRGFHIFPK(0)

[37] [38]

>gi|15354903|gb|BI504529.1| BB170023B10C08.5 Bee Brain Normalized/Subtracted Library, BB17 Apis

mellifera cDNA clone BB170023B10C08 5', mRNA sequence.

Length = 657

[39] [40]

Score = 190 bits (483), Expect = 1e-48

Identities = 88/114 (77%), Positives = 97/114 (85%), Gaps = 7/114 (6%)

Frame = +3

[41] [42]

Query: 18 YYYYSLKYAFWKDRGISGPKPIIFFGNLGNSIIKKKSLSETVKKWYDDYKHESVFGIFEG 77

YYYYS KYAFWKDRGISGPKPI+FFGN GNSI+KK+S+SETVKKWYDDYKHESVFGI+EG

Sbjct: 3 YYYYSSKYAFWKDRGISGPKPIVFFGNFGNSIVKKRSISETVKKWYDDYKHESVFGIYEG 182

[43] [44]

Query: 78 TIPVLVINDLDMIKDILIRDFSLFVDRGFHIFPK-------LLLVDGNLWNEQR 124

TIPVLVINDLDMIKD+LIRDFS+FVDRGFH FPK L L++ W R

Sbjct: 183 TIPVLVINDLDMIKDVLIRDFSIFVDRGFHTFPKIEPLTQHLFLLEAERWRPMR 344

[45] [46]

an EST 95% to 6AS2 above, exact match to Un.601a from ver 1.2

There might be alternative splicing of the first exon.

YYYYSSKYAFWKDRGISGPKPIVFFGNFGNSIVKKRSISETVKKWYDDYKHESVFGIYEG

TIPVLVINDLDMIKDVLIRDFSIFVDRGFHTFPKIEPLTQHLFLLEAERWRPMRMKLSPI

FTSGKLKEMFSLIVESAGNLEKYLDEVIKKNEMVECRDLAAKFTTDVIGSCAFGINTNSL

LEEDSEFRrMGKKIFSPSLKLMIGNTCKVFFPSLYEVI

[47] [48]

>CYP6AS13 Am3_Un.8296 (9442-11518), 497 aa, 5 exons

version 2 = CYP6AS13 CYP6ASf1 Am2_Un.5370 (9442-11518), 497 aa, 5 exons, completion on C-terminus stop codon changed to A (63% to 6AS2)

version 1.2 = CYP6AS fragment Am1.2_Un.1326b incomplete on C-terminus (4-243) minus, 80 aa, 1 exon

1 aa diff to Am1.2_Un.5413

version 1.2 Am1.2_Un.5413 stop codon in 3rd exon, incomplete on both ends, (1-1334) plus, 315 aa, 3 exons 60% to 6AS2, 71% to Am1.2_Un.6932 in one overlapping exon 3

MDFFQIFCAICIMLLAIYYYYTSIYNYWKVRGIPGPEPTIIIGNFMEVFLKKISINDKLRFLYNKYKNEPMFGIFEGSSPILVLNDLDLIKDVLIKDFSIFSNRGFRIFPK(0)AEPLGEHLFALETERWRPMRAKLSPIFTSGKLKEMFPLIIECSKNMEPYLDKIAERGKYIECRDLAAKFTTDVIGSCAFGIDMNSISDKDSEFRIIGRKLFTPTFKTIVRDVCRQFLPGLYDVIGHKLQIEEVNEFLTNLIKDTINYRKENKIVRPDFVNTLIELKDHPEKLETIK(1)LTDSMIASQAFVFFVAGFETSSSTISHALYELAQNQEIQDKLREEIREVYEKHGELTYDVIKNMKYLDKVLKE(1)TLRKYPIMAMLTREAQENYTFKGTKVTIEKGIKVWILPYGIQNDPDIFPNPDIFDPERFDEEAVAARHPMSYLPFGDGPRNCIG(1)ARFAQFQSKIGIITIVRNHKIDVCEQTKIPYESDPFQFLLALKGGINLKISKI*

[49]

>CYP6AS2P1 Am3_Un.8278 (2917-1936), 209 aa, 3 exons, exons 3,4,5

version 2 = CYP6AS2P1 CYP6ASf2 Am2_Un.5353 (2878-1897), 209 aa, 3 exons, lacking expected intron acceptor site on last intron breaking heme motif, frameshift in middle of last exon

version 1.2 = CYP6AS fragment Am1.2_Un.6932 incomplete on N-terminus (41-1021) minus, 209 aa, 3 exons, 92% to last 3 exons of CYP6AS2

(1)LTDILLTAQAVVFFVAGFETSSSTMAFSLYELAQNQEIQDKLREEIRKVHEKNKGVLTYTDIKEMKYLDKVFKE(1)

TLRKYPILSTLSRKVMENYTFKGTKITIPKGTKIWVHGIQHDPNIYPEPEVFDPERFEDDAFASRHPMSYLPFGDGPRNCIG(1)

LVLPHYQSKVGLITILRNHKXVNVCEKTTIPFKADERSFLLVPKGGIHLKIIKI*

[50] [51]

6AS2 (top) compared to 6ASf2

Query: 1 LTDSMIASQAFVFFVAGFETSSSTISHALYELAQNQEIQDKLREEIREVYEKH-GELTYD 59

LTD ++ +QA VFFVAGFETSSST++ +LYELAQNQEIQDKLREEIR+V+EK+ G LTY

Sbjct: 1 LTDILLTAQAVVFFVAGFETSSSTMAFSLYELAQNQEIQDKLREEIRKVHEKNKGVLTYT 60

[52] [53]

Query: 60 VIKNMKYLDKVLKETLRKYPIMAMLTREAQENYTFKGTKVTIEKGIKVWILPYGIQNDPD 119

IK MKYLDKV KETLRKYPI++ L+R+ ENYTFKGTK+TI KG K+W+ +GIQ+DP+

Sbjct: 61 DIKEMKYLDKVFKETLRKYPILSTLSRKVMENYTFKGTKITIPKGTKIWV--HGIQHDPN 118

[54] [55]

Query: 120 IFPNPDIFDPERFDEEAVAARHPMSYLPFGDGPRNCIGARFAQFQSKIGIITIVRNHK-I 178

I+P P++FDPERF+++A A+RHPMSYLPFGDGPRNCIG +QSK+G+ITI+RNHK +

Sbjct: 119 IYPEPEVFDPERFEDDAFASRHPMSYLPFGDGPRNCIGLVLPHYQSKVGLITILRNHKXV 178

[56] [57]

Query: 179 DVCEQTKIPYESDPFQFLLALKGGINLKISKI 210

+VCE+T IP+++D FLL KGGI+LKI KI

Sbjct: 179 NVCEKTTIPFKADERSFLLVPKGGIHLKIIKI 210

[58] [59]

the bottom seq has some defects. The last frameshift X should not be counted as an extra amino acid since it adds to the length and this does not match 6AS2.

[60] [61]

The heme region is out of phase

[62] [63]

1: K C I F D A V F L * I L I L I F V I I F

2: N A F S M Q F F Y K F * F * F S * L F F

3: M H F R C S F F I N F N F N F R N Y F F

AAATGCATTTTCGATGCAGTTTTTTTATAAATTTTAATTTTAATTTTCGTAATTATTTTT

901 ---------!---------!---------!---------!---------!---------! 960