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

TTTACGTAAAAGCTACGTCAAAAAAATATTTAAAATTAAAATTAAAAGCATTAATAAAAA

[64] [65]

[66] [67]

1: F S A R F A A L S K * G W T Y N N S S Q

2: S V L V L P H Y Q S K V G L I T I L R N

3: Q C S F C R I I K V R L D L * Q F F A I

TTCAGTGCTCGTTTTGCCGCATTATCAAAGTAAGGTTGGACTTATAACAATTCTTCGCAA

961 ---------!---------!---------!---------!---------!---------! 1020

AAGTCACGAGCAAAACGGCGTAATAGTTTCATTCCAACCTGAATATTGTTAAGAAGCGTT

[68] [69]

The green show a PROBABLE FRAMESHIFT

[70] [71]

The second small 2 aa deletion is in a region where length is usually conserved, between PKG and PERF motifs. These multiple defects tag this sequence as a pseudogene

[72] [73]

>CYP6AS3 Am3_Un.433 (21903-19550), 501 aa, 5 exons

version 2 = CYP6AS3 Am2_Un.476 (20769-18418), 499 aa, 5 exons

version 1.2 = CYP6AS3 Am1.2_Un.601b (7365-9709) plus, 499 aa, 5 exons

version 1.1 = CYP6AS3 Am1.1_Un.792b (3396-5727) minus, 499 aa, 5 exons

64% to AmGroupUn.5496

MDYFQLLCVIGALLFAIYYYLTLTFDTWKNRGIPGPKPTIFFGNFQEVILKKISLAEKTKQLYQEYKNELVFGIFQGRTPILVINDLEMIKDVLIRDFSVFPDRGIHVNPK(0)VEPIFQTLFSLKSKTWRPLRMKLSPVFTSGKLKDMFPLILDCAKNLEEFVEKVRNSGEPVDCRDMAAKFTTDVIGSCAFGVCMNSLSPEGSEFRRMGEQLGKFSFKKLARDFTRLYMPFLFDIIGGYLQSHEVNNFFINLIRDSIKYRQENNVYRPDFVNTLKELKEHPEKLENIE(1)LTDALLTSQALVFFLAGFETSSTTISNALYELAQNPEMQDKLRKEIKEVYENNGGALSYTDVKEMKYLDKVFKE(1)TLRKYPVLAALSRQATENYTFKDTKIKISKGTRIWIPVYGIQHDPNIYPEPEVFDPERFEDDAFASRHPMTYLPFGDGPRNCIG(1)ARFAHYQSKVGLITILRNNKVEVCAKTLIPYKSEPRNILMIPKGGKVELRITKV*--

[74] [75]

>CYP6AS3frag Am3_Un.2604 (1325-405), 212 aa, 3 exons, exons 3,4,5 from CYP6AS3, except N->D, R->G

(1)LTDALLTSQALVFFLAGFETSSTTISNALYELAQNPEMQDKLRKEIKEVYENNGGALSYTDVKEMKYLDKVFKE(1)TLRKYPVLAALSRQATENYTFKDTKIKISKGTRIWIPVYGIQHDPNIYPEPEVFDPERFEDDAFASRHPMTYLPFGDGPRNCIG(1)ARFAHYQSKVGLITILRDNKVEVCAKTLIPYKSEPRNILMIPKGGKVELGITKV*

>CYP6AS4 Am3_Un.433 (2852-5513), 501 aa, 5 exons

version 2 = CYP6AS4 Am2_Un.476 (1718-4379), 501 aa, 5 exons

version 1.2 = CYP6AS4 Am1.2_Un.1740a (1295-3956) plus, 501 aa, 5 exons

version 1.1 = CYP6AS4 Am1.1_Un.1753 incomplete on C-terminus of exon 1 (2509-4596) minus, 411 aa, 5 exons

80% to AmGroupUn.42b, 58% to AmGroupUn.792b, 41% to 6a17, 44% to 6a13,

MLHHFHILTAFVAIFLALYYYLTSKFDFWKNRGVSGPRPVPFFGNAKDVLLRKIGIGSFIAELY

KRYDNEAMFGIFIGRSPNLVLRDLDLIKDVLIKDFSIFDNRGLNIPER(0)

AEPFSVNLFSVDATRWRPLRMRLSPVFTSGKLKEMFPLILECAEHLEQCLEDAVKRGGPVDCFEIPARYTTDVIGSCAFGINMNALSDERSEFRKMGRNMFDQNMIKFTRNLLRDFFPRFYNLLGFVLPYTESTVFMT

LIKGTIKYREENDVVRPDFVNLLMELKKHPEKLKNIE(1)

ITDTLLAAQASVFFAAGFETSSTTMAHALYEMALNPDIQDKLRNEMKEFHAKNNGNLKYEDIKEMKYLDKVFRE(1)

TLRKYPPGMLLRRKCNSNYTFHGTKVSIPAGTSVIIPLYAIQIDPKFYENPDVF

DPERFNEDAVAARHPMTYLPFGDGPRNCVG(1)

ARFAVYQTKVGLIKILQNFRVDVCEKTMIPYVKKINSITLAPRDGIFLKIEKITD*

[76] [77]

>CYP6AS5 Am3_13.15 (1226-4270), 498 aa, 5 exons, stop codon in exon 4, differences are highlighted

version 2 = CYP6AS5 Am2_Un.4238 (9913-6869), 498 aa, 5 exons, pseudogene?, stop codon is back in 4th exon

version 1.2 = CYP6AS5 Am1.2_13.15b (125743-128770) minus, 499 aa, 5 exons, now complete

version 1.1 = CYP6AS5 Am1.1_Un.42a pseudogene? stop codon in exon 4 before heme binding

site and missing exon 5 (126603-129298) minus, 439 aa, 4 exons

56% to AmGroupUn.5496 43% to 6M2 new subfamily in CYP6?

MASSFEILCGIAVLFLALYYYLTSTFDFWKSRGVVGPKPVPFFGTTKDLILVKKSTAHFVK

DIYEKYKNEPMVGLYATRSPFLLLNDPELIKDILIRDFSKFANRGLGVFER(0)

TEPLSPHLLNLEVERWRPLRSRLSPIFTSGKLKEMFYLIIECSLNLEMYLDKLIEKNEPIECRELTARFTTDVIGSCAFGIDMSSMTNENSEFRRMGREVFAVNFMNVMRMKLKQFMPRLYDLLGYVMPDRTFAPFF

TRVVTDTIKYRNDNNIVRPDFINMLMELQKNPQKLENIK(1)

LTDSLIAAQAFVFFLAGFETSSTTMSNALYELALNQDVQKKLREEINTFCPKNNKELKYDDIKEMEYLDKVFKE(1)

TLRMYPPASILMRKAISDYTFNDTKITIPKEMKIWIPAFAIHRDSAI

YPNPDSFDPERFDKDAMASRHPMHYLPFGDG*RNCIG(1)

ARFAVYQTKVGLITILRNHKVEVCEKTVIPYEFDPGAFLLSPKDGIYLKITKI*

[78] [79]

>CYP6AS6v1 Am3_13.15 (12051-14321), 497 aa, 5 exons, 5 aa differences between CYP6AS6v1 and CYP6ASv2 are highlighted

version 2 = CYP6AS6 Am2_13.15 (124021-121751), 497 aa, 5 exons

version 1.2 = CYP6AS6 Am1.2_13.15c (120924-123194) minus, 497 aa, 5 exons

version 1.1 = CYP6AS6 Am1.1_Un.42b alternate splice for exon 3? (121844-124324) minus,

497 aa, 5 exons

41% to 6a17, best matches all CYP6As 80% to AmGroupUn.1753

MFDYFQILIAFVASFLALYYYLTSNFDFWKNRNVVAPKPIPFFGNTKDVVLKKIEISNFIAELYKKYENEAMFGIFFGGSPNLILRDLDLIKDVLIKDFSTFDERGFKISER(0)ADPLNANLFNMDVTRWRPLRIKLSPVFTSGKLKEMFPLILKCAERLEQCLEDAVKRGGPVDCFEISARYTTDVIGSCAFGINMNALSDERSEFRRIGKRIFDLDKNILRSFLRQFFPRFYNLLGFVIPYSETSKFVTKFISEMIKYREENNVVKADFVNLLMELKKHPEKLQNIK(1)ITDNLLAAQAFVFFAAGFETSSTTMAHALYEMALNPNIQDKLRKEIKEFYANNNFTYEEVKKMKYLDKVFKE(1)TLRKYPPGVFLKRKCNSNYTFKGTKVSIPAGTSVIIPVYSIQTDPKFYENPDVFDPERFNEDAVAARHPMTYLPFGDGPRKCIG(1)IRFGVYQTKVGLIKMLKNFKVNVCEKTMIPYIKKINSFTLAPKDGIFLKIEKNN*

[80] [81]

>CYP6AS6v2 Am3_13.15 (6830-9094), 497 aa, 5 exons

version 2 = CYP6AS6v2 Am2_Un.4238 (4309-2045), 497 aa, 5 exons, completes on N-term what appeared to be a single exon fragment in version 1.2

version 1.2 = CYP6AS6-se1[5] Am1.2_13.15a solo exon pseudogene (132407-132573) minus, 54 aa, 1 exon

1 aa diff to last exon of CYP6AS6, but in different region on 13.15

the two sequences are on AADG03005752.1 11483 bp apart

MFDYFQILIAFVASFLALYYYLTSNFDFWKNRNVVAPKPIPFFGNTKDVVLKKIEISNF

IAELYKKYENEAMFGIFFGGSPNLILRDLDLIKDVLIKDFSTFDERGFKISER(0)

ADPLNANLFNLDVTRWRSLRIKLSPVFTSGKLKEIFPLILKCAERLEQCLEDAVKRGGSVDCFEISARYTTDVIGSCAFGINMNALSDERSEFRRIGKRIFDLDKNILRSFLRQFFPRFYNLLGFVIPYSETSKFVTKFISEMI

KYREENNVVKADFVNLLMELKKHPEKLQNIK(1)

ITDNLLAAQAFVFFAAGFETSSTTMAHALYEMALNPNIQDKLRKEIKEFYANNNFTYEEVKKMKYLDKVFKE(1)

TLRKYPPGVFLKRKCNSNYTFKGTKVSIPAGTSVIIPVYSIQTDPKF

YENPDVFDPERFNEDAVAARHPMTYLPFGDGPRKCIG(1)

IRFGVYQTKVGLIKMLKNFKVNVCEKTMIPYIKKINSFTLAPKDGIFLKVEKNN*

[82]

This has only 5 aa diffs to 6AS6. four are in exon 2. Is it a different gene? are the introns different? Is it an allele?

[83]

intron 1

CYP6AS6v1_i1 GTATGTATTTATTTTTCTTTTATATATATATATATATAAATTTTTTTTTAACATTTCAAA

CYP6AS6v2_i1 GTATGTATTTATTTTTTTTTTATATATATATATA----AATTTTTCTTTAACATTTCAAA

**************** ***************** ******* **************

CYP6AS6v1_i1 ATATAGAAAGTTCAAAATTTTATTAATTTTTTTAGAAAATCCTCTTAAAATAATCTTCTT

CYP6AS6v2_i1 ATATAAAAAGTTCAAAATTTTATTAATTTTTTTAGAAAATCCTCTTAAAATAATCTTCTT

***** ******************************************************

CYP6AS6v1_i1 AAAATAAAGTGTTTCGATTTAATTTTTTAACTATAG

CYP6AS6v2_i1 AAAATAAAGTGTTTCGATTTAATTTTTTAACTATAG

************************************

intron 2

CYP6AS6v1_i2 GTAAGTAAAATAATTTGTAATTTTAATTCTTTCAAAAAGCATAATAAAAGCATAAATAAA

CYP6AS6v2_i2 GTAAGTAAAATAATTTGTAATTTTAATTTTTTCAAAAAGCATAATAAAAGCATAAATAAA

**************************** *******************************

CYP6AS6v1_i2 GTTGTAGTTTATGATTATTTCATAAATAGAATTTAAAGATGTATAATGATGAAATAAAAT

CYP6AS6v2_i2 GTTGTAGTCTATGATTATTTCATAAATAGAATTTAAAGATGTATAATGATGAAATAAAAT

******** ***************************************************

CYP6AS6v1_i2 TTGAAATAGAATTTTAAATTTATAAATTTTTTGTCTAATGTTTTATAAATAATTAATAAA

CYP6AS6v2_i2 TTGAAATAGAATTTTAAATTTATAAATTTTTTGTCTAATGTTTTATAAATAATTAATAAA

************************************************************

CYP6AS6v1_i2 ATATTATATTAATGTATTATTAAAATACTATTTTTTAATAAAATTTTACAG

CYP6AS6v2_i2 ATATTATATTAATGTATTATTAAAATACTATTTTTTAATAAAATTTTACAG

***************************************************

intron 3

CYP6AS6v1_i3 GTATATTTTTTAAACTTATTTTAACAAAATGTATAAAAAATGTAAAATTTTTTATACATT

CYP6AS6v2_i3 GTATATTTTTTAAACTTATTTTAACAAAATGTATAAAAAATGTAAAATTTTTTATACATT

************************************************************

CYP6AS6v1_i3 ATTATTAATATTCTATTATATATATATTTTTATATATATTTTTGCAAATAATTAGAAAAC

CYP6AS6v2_i3 ATTATTAATATTCTATTATATATATATTTTTATATATATTTTTGCAAATAACTAGAAAAC

*************************************************** ********

CYP6AS6v1_i3 CAATAATTTATTATTATGTATAAATTATCATATTGAAATATGTGTAAAGATTATAATTTA

CYP6AS6v2_i3 CAATAATTTATTATTATGTATAAATTATCATATTGAAATATGTGTAAAGATTATAATTTA

************************************************************

CYP6AS6v1_i3 AAATTATATTATTTTAAATAAGATAATTATTACTTTAAAATAAAATTATATTTATCTGTC

CYP6AS6v2_i3 AAATTATATTATTTTAAATAAGATAATTATTAC--TAAAATAAAATTATATTTATCTGTC

********************************* *************************

CYP6AS6v1_i3 TTTTTAAATAG

CYP6AS6v2_i3 TTTTTAAATAG

***********

intron 4

CYP6AS6v1_i4 GTAATTCAAAGAAAAAATACTAAAATGTTTTTTAGATGATAATGATAATTTGTATCTTAC

CYP6AS6v2_i4 GTAATTCAAAGAAAAAATACTAAAATGTTTTTTAGATGATAATGATAATTTGTATCTTAC

************************************************************

CYP6AS6v1_i4 GATTATTAAAATTTTTAACATTTACTTGTTTTATATTTTTATATATTAATGATAATCAAT

CYP6AS6v2_i4 GATTATTAAAATTTTTAACATTTACTTGTTTTATATTTTTATATATTAATGATAATCAAT

************************************************************

CYP6AS6v1_i4 GATTAAATTATTGTTTCAG

CYP6AS6v2_i4 GATTAAATTATTGTTTCAG

*******************

>CYP6AS7 Am3_13.15 (16943-19515), 505 aa, 5 exons

version 2 = CYP6AS7 Am2_13.15 (119129-116557), 505 aa, 5 exons, completed on N-terminus

version 1.2 = CYP6AS7 Am1.2_13.15d (115784-118302) minus, 487 aa, 5 exons AADG03005751.1 (WGS)

version 1.1 = CYP6AS7 Am1.1_Un.42c incomplete on N-terminus (115419-117055) minus, 265 aa, 3 exons

45% to 6M2, 66% to AmGroupUn.1753 Nearly complete sequence.

BI505169.1 EST (N-terminal)

MPSLEIIFGIILLFPAIYYYFTSTFDFWKVRGVPGPKPIPIFGNIKNVMLLKTSMCHYLKKLCEEYKHEPMIGIFTRKTPILIIQDPDLIKDVLIRDFSKFANRGIPIHEK(0)AEPLSPHLFNLEVERWRPLRTRLSPVFTSGKLKEMFPLILDCAKHLEQYLDKLVLREEFIECRELTAKYTTDVIGSCAFGIEMNALSDEESEFRRIGRKVFSNSFGQILRFRFRQIFPRIYNLLGFVLPPMEVTKFLTNIIVSTMKYRQENNIVRPDFVNMLIELKKHPDKLENIK(1)LTDTLLTAQAFVFFIAGFETSSSAISNALYELALNPEVQNKLRQEIKEYFNKHNELKYEYIKNMIYLDLVFRE(1)TLRKYPPGPLILRKSITNYTFNNTKVSIPEESFVWIPLYAIHHDPKIYPNPDAFIPERFNDDAIATRHPMHYLPFGDGPRNCIG(1)ARFAVYQSKIGLITILWNYKVEVCDKTMIPYEINPAAFLLTPKGGIYLKFTKIKNNEEILN*

[84] [85]

>CYP6AS8 Am3_13.14 (155751-158416), 500 aa, 5 exons, alternate first exon 2000nt upstream

version 2 = CYP6AS8 Am2_13.16 (153493-156158), 500 aa, 5 exons

version 1.2 = CYP6AS8 Am1.2_Un.5081 (10839-13505) minus, 500 aa, 5 exons version 1.1 = CYP6AS8 Am1.1_Un.4533 (10741-13406) minus, 500 aa, 5 exons

53% to AmGroupUn.2631 42% to 6P4 48% to 6N1 partial

MYISLEIFCGIVVALIALYYYLTVNNNFWKNRGIAGPEPVLGFGNMKKVLLGKESMSQFLTKIYHEYKNEPIIGIFTTRTPQLIIKDPDLIKTILIKDFSKIMNRGLLPMVS(0)GEPISQHLFNIEAERWRPLRIHLTPVFTANKLRGMFSLILECSMHFVSYVDSLVKKGEPVNVREVAARFTTDVVGSCGFGVEMNSLSEKESEFRRVGKSVFATNYARIIKHRIREFMPRLYNYILYLWPTDEMAEKIIKLTRETLEYREKNNLFRPDFMNILLDLKKHPEKIGLD(1)VTNEFLAAQAFIFFVAGFETSSSTISNALYELALNPDVQDKLRKEIKEFAAKNDGEWRYETIKEMEYLGKVFQE(1)TLRKYPSLPFLTRELIEDYTFESNKVTIPKGLKIWIPTYAIHNDPDIYPDPDKFDPERFSDDKIKQRHPMHFLPFGHGPRNCIG(1)ARFAIYQTKIGLINILRNFKLDVCDKTLIPYKHHPRGLLLMPLTDLYLKITRLTN*

[86] [87]

>CYP6AS9 Am3_13.16 (7981-5161), 499 aa, 5 exons, single nucleotide insertion in 2nd exon (at X); 3rd intron has no good phase 1 donor; 4th intron donor is 'GC'

version 2 = CYP6AS9 Am2_13.17 (7981-5161), 497 aa, 5 exons, probably pseudogene, frameshift in 2nd exon, no good splice donor site for 3rd exon, missing IG from heme binding region – there are a 2 fragments that are similar to CYP6AS9

version 1.2 = CYP6AS9 Am1.2_Un.1054b frameshift in exon 2, possibly a pseudogene because splice site donor for intron 4 removes 2 aa from heme binding region (1823-4511) minus, 499 aa, 5 exons

version 1.1 = CYP6AS9 Am1.1_Un.2792 incomplete on N-terminus, pseudogene??? heme binding

site probably not functional due to change in splice donor

(995-3268) minus, 367 aa, 4 exons

65% to 6AS8

Am1.1_Un.4458 incomplete on both ends (1-1218) plus, 203 aa, 2 exons

3aa diffs to AmGroupUn.2792 61% to 6AS8

note: GG (9) boundary was QE (1) in ver 1.2

MFINLETLCGFVIVLIAFYYYLTINNNFWKNRGIPGPKPTIGFGNMWTVMFGKESFSQLLTTIYNKYKDEPMIGIFFRRRPVLLLKDFDLIKDVLIKDFSKFANRGFLKTNP(0)KVPLTNHLFALEVKRWRPLRNHXSSPVFTSGKLKGTFAQILNCSNDLVTHIDTLSKMEDSINMREVAAKFTTDVIGSCVFGIKMNSLSEIENEFRRIGRNIFATNFTNILKIRLLQSTPFLHSLLCKILPDEEMEMFMRITKETMEYREKNNLVRPDFMNILLELKKHPERIADIE(1)LTDDLLAAQAYIFFAAGFETSSTTISNALYELALNHDMQDKLREEIKEFEAKNDGEWRYETIKEMQYLGKIFGG(9)TLRKYPALSFLSRESIIDDYTFRDTKLTISKGTLVWIPVFPIHHDPNIYPDPEKFDPERFTEDKIKERNLMHYFPFGHGPRNCIG(1)ARFAIYQTKIGLIKILRNYKVDVCSKTLIPYKYDPFSFILVPLGGLYLKITRI*

[88] [89]

>Am2_79040249 Am3_79040249 (235-735), 169 aa, 1 exon

version 2 =Am2_79040249 new (255-761), 168 aa, 1 exon highly similar to 6AS9

(0)KVPLTNHLFALEVKRWRPLRNHLSPVFTSGKLKGTFAQILN?APMILXDSYRHFIEDGRLYKHAREVAAKFTTDVIGSCVFGIKMNSLAEREEFRRIGRNIFATNFTNILKIRLLTSTPLLHSLLCRILPDEEMRMFFRITKETMEYREKNNPPQAGFYEYTSRLETEV

[90] [91]

>Am2_Un5369 Am3_Un.8295 (5212-3283), 302 aa, 4 exons, exons 2,3,4,5 of gene similar to CYP6AS9; frameshift in first exon at X; 'GC' spice donor on final intron adds 'IG' to heme motif;

version 2 = Am2_Un.5369 (5212-3283), 300 aa, 4 exons C-terminal end of Am1.2_6AS9, also appears to be a pseudogene

version 1.2 = NUn.5369 UnOriented 6AS?? pseudogene

EIENEFRRIGRNIFATPFYKYIEDXRLLQSTPLLHSLLCKILPDEEMEMFMRITKETMEYREKNNLVRPDFMNILLELKKHPERIADIE(1)ITDDLLAAQAYIFFAAGFETSSTTISNALYELALNHDMQDKLREEIKEFEAKNDGEWRYETIKEMQYLGKIFQE(1)TLRKYPALSFLSRESIIDDYTFRDTKLTISKGTLVWIPVFPIHHDPNIYPEPEKFDPERFTEDKIKERNLMHYFPFGHGPRNCIG(1)ARFAIYQTKIGLIKILRNYKVDVCSKTLIPYKYDPFSFILVPLGGLYLKITRIQD*

[92] [93]

>Am2_48488186 Am3_48488186 (826-453), 125 aa, 1 exon, single nucelotide deletion at X, causing frameshift

version 2 = Am2_48488186 like 2nd exon of Am1.2_6AS9 (826-453), 125 aa, 1 exon

DTLSKMEDSINMREVAAKFTTDVIGXSCVFGIKMNSLSEIENEFRRIGRNIFATNFTNILKIRLLQSTPFLHSLLCKILPDEEMEMFMRITKETMEYREKNNLVRPDFMNILLELKKHPERIADIG(1)

[94] [95]

>CYP6AS10 Am3_13.16 (13740-10480), 499 aa, 5 exons

version 2 = CYP6AS10 Am2_13.17 (13740-10480), 499 aa, 5 exons

version 1.2 = CYP6AS10 Am1.2_Un.1054a (7016-10145) minus, 499 aa, 5 exons

version 1.2 = CYP6AS10 Am1.1_Un.2631 incomplete on C-terminus of exon 1 (2686-5804)

minus, 495 aa, 5 exons AADG03012613.1 AADG03012614.1

gnl|Amellifera1|174017188 BCM Apis mellifera 12/11/2003

40% to 6M2 53% to AmGroupUn.6AS8

MAAFEILCGFIIFIFAFYYYLIKPQEYWKNRGVPGPKPIPIFGNFFRLTFARISIGDLMTKFYKEYKHEPVFGLYMRNVRVLAINNPDLIKTVLIKDFSKFAHRGLALNEV(0)TEPLSQHLFVLEPKRWRPLRTKLSPIFTSGKLKDMFSLIIECSNTLENYVEHLISKNDRVEVRDLAAKFTTDVIGSCGFGVDMNAMSDVQCKFRDIGREFFGPSFKQILKIRLRENLPRLYTFLGYILPRDETTTFFTNVVLDMIKYRKTNDIYRPDFINALINIQNHPEKLDIE(1)LTEPLLVAQAFLFFVAGFETSSLTIATALYELAQNQDIQDKLRDEITEHHKLNNGEWQYENIKNMPYLDAVFKE(1)TLRKYVPLTVLMRQSLEDYTFESINLTIPKDTRIFIPIYAIHRDPDIYPNPEVFDINRFSKEAEATRHPMHYLPFGDGPRNCIG(1)ARFAIFQTKIGLIKILRTYKVDVCNETQIPFINEPRTFTLAPKHDLTLKITKIEN*

[96] [97]

>CYP6AS11 Am3_13.14 (148342-151323), 500 aa, 5 exons

version 2 = CYP6AS11 Am2_13.16 (147797-150778), 500 aa, 5 exons, not a pseudogene after all??? complete on N-terminus, 11 aa changes

version 1.2 = CYP6AS11P (pseudogene) Am1.2_Un.5080a incomplete on N-terminus (1-1697) plus, 229 aa,

4 exons with three frameshifts and a stop codon

gnl|Amellifera1|166350743 BCM Apis mellifera 12/11/2003

Am1.1_Un.4532 incomplete on N-terminus (1-757) minus, 131 aa, 2 exons

83% to 6AS8

MYINLEIFCAIVIAFIAFYYYLTINNNFWKHRGISGPKPVLGFGNMKKIILGEESMSQYLTKLYHKYKNESMIGIFRLRTPALIIKDPDLIKIVLIKDFSKFMNRGLLPIIS(0)TEPISHHLFALEAERWHPLRKHLTSGFTSNKLKGMFCMIHECSKHLVNYLDILVRKEEPVNVREVAARFTTDVVGSCGFGVEMNSLSEQESEFRRLGKSIFNTNVQKIIKDRIRELTPQVYNFLLYILPLDGISPKILKLMKETIKYRKKYDIFRPDFMNIILELKKHPEKINID(1)ITNELLAAQIFIFFAAGFETSSTLISNALYELALNPNIQDKLREEIKKFESQNDEEWKYETIKKMDYLEKVIQE(1)TLRKYPPVPFLNRELIDDYTFESNKVTIPKGLKIWIPTYAIHNDPDIYPDPDKFDPERFSEDNIKQRHPMHFLPFGHGPRNCIG(1)IRFAEYQTKIGLINILRNFKLDVCDKTLIPYKLHPRGLILIPLTDLYLKITRLTN*

[98] [99]

>CYP6AS11alt Am3_13.14 (152857-153192), 112 aa, 1 exon, exon 1 from CYP6AS11, but 2000nt upstream from start of CYP6AS8

version 2 = CYP6AS11alt Am2_13.16 (152312-152647), 112 aa, 1 exon this sequence is just upstream from the start of CYP6AS8 (about 2000bp) or about 1500 bp from 6AS11

version 1.2 = Am1.2_Un.5080b incomplete on C-terminus (3231-4470) plus, 287 aa, 2 exons

version 1.1 = Am1.1_Un.248b incomplete on C-terminus and N-terminus of exon 2

only 6 amino acid differences to 6AS11P in an 11 amino acid window

MYIGFEIIYGIVIVFIAFYYYLTINNNFWKHRGISGPKPVLGFGNMKKIILGEESMSQYLTKLYHKYKNESMIGIFRLRTPALIIKDPDLIKIVLIKDFSKFMNRGLLPIIS(0)

[100] [101]

>CYP6AS11frag Am3_Un.6498 (653-131), 175 aa, 1 exon, exon 2 only

(0)TEPISHHLFALEAERWHPLRKHLTSGFTSNKLKGMFCMIHECSKHLVNYLDNLVRKEEPVNVREVAAKFTTDVVGNCGFGVEMNSLSEKESEFRRLGKSIFNTNLQKIIKDRIRELTPQIYNFLLYILPLDGISPKILKLMEETIKYRKKYDIFRPDFMNIILELKKHPEKINIG(1)

>CYP6BC1 Am3_1.17 (861221-865384), 476 aa, 5 exons

version 2 = CYP6BC1 383+1.19 Am2_1.19 (858065-862228), 476 aa, 5 exons, still incomplete on N-term of 4th exon, combines Am1.2_Un.383 and Am1.2_1.19 about 46% to 6AS7

version 1.2 = Am1.2_1.19, incomplete on C-terminus (860909-863534) plus, 381 aa, 3 exons

version 1.2 = Am1.2_Un.383 incomplete on N-terminus (6360-7086) plus, 84 aa, 2 exons

version 1.1 = Am1.1_Un.4500 incomplete on N-terminus (298-1025) minus, 84 aa, 2 exons

MWNIIRELLEQFLLPGLFLGILYCFLTSTFDFWKNRGVPFRKPTVLFGNFAPMLLFRKSLPEGIKEMYEWFKDERYFGAFRVRSPVLILRDPDLVKNICVKNFTSFSNRGIPVNSQ(0)DPLSAHLFNLEGKKWKSLRSKLTPAFSSGKLKRMFYLLAECGEEFEKLIDISSETDRPYEIRELAAKFTIDVIGTCAFGIQINALTDEESEFHRAAKKLSKPSYKATLWRMLRTAMPRLYKFLGVQVIDPGVTKFFKDVVSQMIKQRGEYGIKRHDFMDLLIELKNKGTLDEFGK(1)LDENSIAAQAFVFFAAGYETSSNTIAFCLHELALNTEIQEKTRRDIQDAIDSRNGNLTYDAVQDMKYLDMVIAG(1)KVELPAGIRVIIPIYGLHHDPDYYPSPAIFNPERFTEENKRTRHPYAYLPFGEGPRNCIG(1)MRFALLQIKVGIISFLRNHRVETCQKTITPIKFSRRSLVTTSEKGFWLRIK*

[102] [103]

>CYP6BC1-like Am3_Un.5223 (4-1346), 141 aa, 2 exons, C-term of exon 3 and C-term of exon 4 from CYP6BC1

YETSSNTIAFCLHELALNTEIQEKTRRDIQDAIDSRNGNLTYDAVQDMKYLDMVIAE(1)TLRKYPPASMLSRRCEYQYQIPNSKVELPAGIRVIIPIYGLHHDPDYYPSPAIFNPERFTEENKRTRHPYAYLPFGEGPRNCIG(1)

[104]

>CYP6AS14 Am3_13.14 (143339-146699), 501 aa, 5 exons

version = CYP6AS14 13.16 Am2_13.16 (142794-146154), 501 aa, 5 exons, complete on C-terminus, -->K

version 1.2 = Am1.2_13.16 incomplete on C-terminus (2365-4885) minus, 445 aa, 4 exons

version 1.1 = Am1.1_Un.248a incomplete on C-terminus (38475-40756) plus, 338 aa, 4 exons 49% to 6Aa14, 73% to 6AS8

MYIGLEILCGIVITLIAFYCYLTINNNYWKNRGIPGPKPVPGFGNMKNVIFGKESVSQFLTRMYNEYKDEPMIGVFSKRTPVLIVKDVDLIKTILIKEFPKFANRGLFPIFS(0)RDPLTHHLFNLEVERWKPLRTQFTPLFTSSKLKEMFSLILECSNHLESYMDTLIKKGEPIDMREVSARYTTDVVGSCAFGIDMNSLSEKESVFRRLGKLIFATNLRKILSIRIQDMLPWLYNSFLYVLPRDEKTRIIMKLMTETMEYREENNVFRPDFINMLLNLKKHPEKIDIE(1)LTDDLLAAQIFIFFAAGFETSSSTISNALYELALNPDIQEKLRKEIKEFEARNNGEWRYEIMKEMEYLEKVFQE(1)TLRKYPSLPFLNRKLINDYTFESNNVTVSKDLKIWIPVYGIHHDPDIYPDPEKFDPERFSKKEEIMKRHPMHFLPFGHGPRNCIG(1)ARFAVYQTKIGLIKILRNFEVQVCNKTLIPYKVNPYTSLLIPITGLYLNVVKLEN*

[105] [106]

>Am2_101397060 Am3_101397060 (245-421), 58 aa, 1 exon, like 6AS14

version 2 = Am2_101397060 (301-576), 91 aa, 1 exon very like Am2_13.16, 29aa in a row

(0)TLRKYPSLPFLNRKLINDYTFESNNVTVSQRFKRFWIPVYRNLTPISDIYPDPEKSHL*KISLGGNYGEGAPKAFLPFGQGPQNMSCVWPIK(0)

[107] [108]

>CYP6AS15 Am3_Un.462 (2742-6273), 501 aa, 5 exons

version 2 = CYP6AS15 218+7762 Am2_Un.210 (2742-6273), 501 aa, 5 exons, combines Am1.2_Un.218 and Am1.2_Un.7762 68% to 6AS8

MNISLEILCGIIVALIVFYYYLIINNNFWKNRKISGPKPVIGFGNMLSIILGKESTSQFLTRIYNEYKNEPMIGIFSKNNPALVIKNPDLIKTVLIKDFHKFANRGLFPVNS(0)REPLSQNLFGLEVERWRPLRIHFSPIFTTNKLKGLCSLILECSEQLEKYMDILIRKGEPLDIREIAARFTTDVIGSCAFGIEMNSLSENESEFRRLGKGVFNTTFRRIVKTRIRNLMPWLYNFFLRILPWDEITKKIVKLTTETIEYRNKNNIVRSDFINVLLNLKKHPEKIAEIE(1)LTNDLLSAQTFVFFGAGFETSSTTISNALYELALNHDIQYKLREEIKEFEKKNDGKWTYESIKEMQYLNKIFQE(1)TLRKYPVVPFLNRELISDYTFENSKITIPKGLKIWIPVYGIHHDPDIYPNPEKFDPERFSEDKIKERHSMHYLPFGHGPRNCIG(1)SRFGTYQTKIGLVKIIRKYKVEICDKTLIPYKFNSFANFLMPSTGLYLMITDVEN*

[109] [110]

>CYP6AS12P Am3_Un.433 (10745-7928), 509 aa, 5 exons highlighted is different from N-term of 2^nd exon in version 2, but similar to N-term of exon 2 in Am2_Un.6006

version 2 = CYP6AS12 Am2_Un.476 (9611-6794), 499 aa, 5 exons, no longer a pseudogene??? complete on C-terminus, 4 aa changes including *->L

version 1.2 = CYP6AS12P Am1.2_Un.1740b possible psuedogene (6374-9226) minus, 500 aa, 5 exons AADG03014312.1

version 1.1 = Am1.1_Un.8712 incomplete on N-terminus (13-1433) minus, 267 aa, 4 exons

60% to 6AS5

version 1.1 = Am1.1_Un.10510 incomplete on both ends (122-647) minus, 176 aa, 1 exon

54% to AmGroupUn.42a, 39% to 6a16m all best hits = CYP6As from aa107-274

probably a new CYP6 subfam or CYP6a

version 1.1 = Am1.1_Un.6966 incomplete on C-terminus (878-1100) minus, 72 aa, 1 exon

40% to 28A5 N-term 38% top 6a20 57% to AmGroupUn.42a

MAYIEILCGIIVSMLVFYYYFTSVFNFWKIRGIPGPKPKFLFGNIRDIILSRISTPTFIKNIYDTYTNEPMVGLYMGRNPILLLKDPELIKDVLIRDFSKFADRGFNVHEK(1)IENFDKFCSQVEPLSQHLFNLEPKRWRPLRSKLSPMFTSKKLKEMFGLILECGHHFEKYVDGLAARRQPVDFCEVAAKYTTDVIGSCAFGINMNAMSSEGSEFREAGRKIFEPTWNSIIRLKFKITMPTLYDLLGPLVPEREVTPFFIKVVTDAMKYRKERNVFRPDFIDTLMKLRDDPESLSDIE(1)LTDAFLTAQAYVFFAAGFETGASTISNTLYELAQNQGMQDRLREEIREHCDKYGGELMYENIKEMEYLDKVFKE(1)TLRKYPPGTLIPRRSVSEYTFKNTNVTIPKGTMIWIPAFPIHRDPNIYPNPDDFNPENFTEDAINNRHPMNYLAFSNGPRNCIG(1)ARFANYQVKIGLIMILRNYKVEVCEKTVIPYQFDPNLFLLGPKGGIYLRVTKVE*

[111] [112]

>Am2_6006 Am3_Un.8658 (2537--56), 396 aa, 5 exons, missing N-terminus of exon 1; single nucleotide deletion at X in exon 1; finished C-term with trace 602155142; similar to CYP6AS12

version 2 = Am2_Un.6006 (2537-189), 375 aa, 4 exons, like 6AS12, but with nucleotide deletion at X in first exon and missing both ends probably the same sequence

YGRIVHGTESDSLLKDPELIKDVLIRXFSKFADRGFNVHEK(0)VEPLSQHLFNLEPKRWRPLRSKLSPMFTSKKLKEMFGLILECGRHFEKYVDGLAARRQPVDFCEVAAKYTTDVIGSCAFGINMNAMSSEGSEFREAGRKIFEPTWNSIIRLKFKITMPTLYDLLGPLVPEREVTPFFIKVVTDAMKYKKESNVFRPDFIDTLMKLRDDPESLSDIE(1)LTDAFLTAQAYVFFAAGFETGASTISNTLYELAQNQGMQDRLREEIREHCDKYGGELMYENIKEMEYLDKVFKE(1)TLRKYPPGTLIPRRSVSEYTFKNTNVTIPKGTMIWIPAFPIHRDPDIYPNPDDFNPENFTEDAINNRHPMNYLAFSNGPRNCIG(1)ARFATISQIVYMISYTVSCEKL*

trace extension:

>602155142 name:107292153 mates:602138461,602160527

ATTTTGTTCCTAGGTGCACGCTTCGCTACTATCAGTCAGATAGTTTATATGATCTCGTATACAGTGAGTTGCGAGAAACTGTGAT

-----1-------- A R F A T I S Q I V Y M I S Y T V S C E K L *

AATTTTGTTCCTAGGTGCACGCTTCGCT <--- Am3_Un.8658

F C S * V H A S

N F V P R C T L R

I L F L G A R F A

>98681617 Am3_98681617 (876-543), 111 aa, 1 exon, frameshift at X, added 12 nucleotides to N-term

version 2 = Am2_98681617 new (840-543), 99 aa, 1 exon 91% to 6AS12P

MAYVKSYAWNHRVDVAFYYYFTSALNFWKIRGIPGPHXPKFLFGNIRDIILSRISTPTFIKNIYDTYTNEPMVGLYMGRNPILLLKDPELIKDVLIRDFSKFADRGFNVHEK(0)

[113]

>CYP6AS16 Am3_Un.8296 (792-4695), 500 aa, 5 exons, single nucleotide deletion in first exon at X, changed donor site of first intron to phase 0 to match other CYP6AS genes

version 2 = CYP6AS16 1326a Am2_Un.5370 (792-4695), 499 aa, 5 exons, completes 3rd exon for Am1.2_1326a, frameshift at X (64% to 6AS9)

version 1.2 = Am1.2_Un.1326a incomplete, missing 3rd exon (4844-8009) minus, 430 aa, 4 exons

version 1.1 = Am1.1_Un.2707 incomplete missing exon 3

(1-3597) plus, 368 aa, 4 exons

59% to AmGroupUn.4533

note XM_396006.1 mRNA predicted by program GNOMON skips exon 3

MIANLEIFCGIIVILIAFYYYIRLEQFWKXIRGIPGPEPLPGFGNVLMIVLGKEAPFQFLTRVYNEFKNEALIGVFMKTYPALVVKDPDLIKDIMIKDFYKFPNRGFPKSDS(0)ADPLTQHLFLVEEEKWRPLRTQLSPVFSTGKLRGTFTQILDCSNHLVTYMDKLVEIGEPIDVREVTAKFTTDVIGSCVFGIKMNSLSGKESEFRRFGRQIFAMNFLKILRLRIKQFLPMLHYLLVRILPPDEETKIMLKLTRDTFKFREAHNIVRPDFMNILMELKKHPEKVPSLD(1)LTDGLLAAQAFIFFAAGFETSSTTVANTLYELALNPDIQEKLRQEIKEFEANNDGEWKYETIQEMKYLDKTFKE(1)TLRKYPVLPYLSRRSIEDYTFEGTKVSIPKNTLICIPVYPIHHDSSIYPNPEKFDPERFSEDEVKKRHSMHYFPFGHGPRNCIG(1)LRFAIYQSKIALIKILSNYKIEICDKTLIPYKYDPFSFISLPLTGIFLKITKLQN*

[114] [115]

>CYP6AS16-like Am3_Un.8276 (1387--287), 213 aa, 3 exons, exons 3,4,5 from CYP6AS16; final exon from trace 174045338

(1)LTDGLLAAQAFIFFAAGFETSSTTVANTLYELALNPDIQEKLRQEIKEFEANNDGEWKYETIQEMKYLDKTFKE(1)TLRKYPVLPYLSRRSIEDYTFEGTKVSIPKNTLICIPVYPIHHDSSIYPNPEKFDPERFSEDEVKKRHSMHYFPFGHGPRNCIG(1)LRFAIYQSKIALIKILSNYKIEICDKTLIPYKYDPFSFISLPLTGIFLKITKLQN*

[116]

>CYP6AS16-2 Am3_13.16 (752-1), 147 aa, 2 exons, like CYP6AS16, except lacking frameshift in first exon, incomplete on C-term from end of second exon, single nucleotide deletion in 2nd exon

version 2 = Am2_13.17 (752-9), 143 aa, 2 exons, like Am1.2_Un.1326a, but lacking the frameshift and missing C-terminus

MIANLEIFCGIIVIVIAFYYYITARNNFWKIRGIPGPEPLPGFGNVLMIVLGKEAPFQFLTRVYNEFKNEALIGVFMKTYPALVVKDPDLIKDIMIKDFYKFPNRGFPKSDS(0)ADPLTKHLFLVEEEKWRPLRTQLXSPVFSTGKLRGT

[117] [118]

>CYP6BD1 Am3_11.14 (431199-428650), 500 aa, 6 exons, 22nt on 3' from repeated sequence downstream used to fill unknown nucleotides, adding 5 aa to C-term

version 2 = CYP6BD1 Un.416 Am2_11.13 (431584-429035), 515 aa, 6 exons, missing aa on C-terminus, changed splice acceptor site on 2nd intron, OK 5aa changes,

version 1.2 = Am1.2_Un.416 incomplete on both ends (10974-18858) minus, 223 aa, 3 exons

gnl|Amellifera1|236106554 BCM Apis mellifera 12/11/2003 42% to 6AQ1

Am1.1_Un.2162 incomplete on both ends (1-852) plus, 223 aa 3 exons

Note there is a seq duplication of the last exon

I used the end of the duplicated sequence to finish the protein sequence.

429191 ARIGQLQSIIGLITIIKNYEISFISKCKGDLDVRNIFITPINDFSL 429054

and 429000 ISLNSKCKGDLDVRNIFITPINDFSLNLTKI* 428905

39% to 6g1 aa175-381 probable new subfamily in CYP6 41% to 6AQ1

MQNSLFMPIIFSIAFFVIILILYLKYTRTYWLRRGVPTVPGHWLFGNIKKILDLKKPPAYVISDIYQKCSENDDILGIYIFFKPFLLIKDPAIIKQILIKDFNYFPDRNFTIQSFYDEIGNKSLFTLKNPQWKYLR(2)TKLSPIFSSAKVKKLFHLMVEAANSMNKYLDDEFSNDTKTKTIMIKDVTLKYTTNVISSVAFGIQVNSFNPKTIQFYEEA(1)QKGLKTTFSRSMQLCISFFFPKLSPYLNTRMLGSSTNFFRKVFWNSMDNREITKTKREDLIDSLMELKNAKQDKDFK(1)FEGDALLSQSAIFFIAGRETSISIICLTLYELAKHPEIQKRTREEINEKLKEHGMTYEGVQSMKYLHQVVSEILRIYPPTPIIDRVAVADYK(0)IPGTDIVIEKGTSVFIVLTALHNDPKYHPDPLRFNPDRFSDENKENIKPFTYIPFGEGPRICIG(1)ARIGQLQSIIGLITIIKNYEISFISKCKGDLDVRNIFITPINDFSLNLTKI*

>CYP6BE1 Am3_Un.1006 (577-5116), 512 aa, 6 exons

version 2 = CYP6BE1 Un.6686 Am2_Un.1086 (577-5116), 512 aa, 6 exons, complete on N-terminus 42% to 6AS2

version 1.2 = Am1.2_Un.6686 incomplete on N-terminus (2630-5871) minus, 453 aa, 5 exons missing about 54 aa

gnl|Amellifera1|2049199734 BCM Apis mellifera 8/15/2003

Am1.1_Un.2637 incomplete on C-terminus (7-2478) plus, 330 aa, 4 exons

45% to 6P4 new subfamily in CYP6? 46% to 6F1

Am1.1_Un.5730 incomplete on both ends (990-741) minus, 84 aa, 1 exon

54% to 6d4m 60% to AmGroupUn.8460 60% to 6AM1

MFLTTWLIPDIIAVASLITVGLYFYYKLYLFKFWHKKGIFYVKPSFPTGNIMPIINGKLSLA(1)EFFRDIYEHNKHHRLVGIYMLYKPYLIVNDPNLIRDILTKEFTNFHDRGIFYNEEVDPLSGHLFQLPGKKWRNLRVK

LTPTFTSGKIKQFFPILNEAGNILAKYLEEEARKGSTIDVKDIFAR(2)

YSTDIIMSVAFGISCDSFKEPNNEFRYWGKKIFDPKPLWNALILFAPQILNFFS

ISYTEKSVTKFFTNMFKQTVKYRESNNIERKDFLNLLIQLMKNGYVDADDESLSNNVNAAK(1)

NKLTMMEAAAQAYVFFLAGFETSSTTVTFCLYELAKNQDIQNKVREEIQTMIKKNGDLTYNALNDMNYLHKVISE(1)

TLRKYPPVVILNRICTNDVKLSTTDFCIPKGTCIAIPVFGLHRDSNI

FPNPEKFDPERFSEENIKTRHPYVYLPFGEGPRICIG(1)

LRFGLIQTKIAIINALLKNKFKFGPNTPSTLEFEKGSLILIGKGGIHLNIEPI*

[119] [120]

>Am2_98611857 Am3_98611857 (16-745), 90 aa, 2 exons, frameshift at X, change in aa around intron

PRDGSSQEKFDPERFSEENIKXLGHPEVYLPFGEGPRI(9)LRFGLIQTKIAIINALLKNKFKFRPNTPSTSEFEKGSLILIGTGGIHLNIEPL*

version 2 = Am2_98611857 new (16-745), 93 aa, 2 exons, 89% to Am1.2_Un.6686, CYP6P like

PRDGSSQE

KFDPERFSEENIKXLGHPEVYLPFGEGPRIL(9)IGLRFGLIQTKIAIINALLKNKFKFRPNTPSTSEFEKGSLILIGTGGIHLNIEPL*

[121] [122]

New families in the 3 clan CYP335 and CYP336

>CYP9P1 name changed from CYP335A1 Am3_14.8 (435001-436548), 515 aa, 1 exon

version 2 = CYP335A1 Am2_14.6 (434001-435548), 515 aa, 1 exon

version 1.2 = CYP335A1 Am1.2_14.13a (10454-12001) plus, 515 aa, 1 exon

version 1.1 = CYP335A1 Am1.1_14.7a (400127-401674) minus, 515 aa, 1 exon

40% to 9f2m new subfamily in CYP9 63% to 9P2 (335A2) 39% to 9D1, 38% to 9K1

50% to 9J8 partial

MEVSHLTTFELLLLTLIFIILAKLVSILYTQFTYWKRNKVPYIRSSPLFGTAWRVFFRLVSFPNYCKYIYNYYPDARYVGVMDFATPTVIVRDPKLIKEIAVKNFNNFPDHRSFVTEEMDPVFGKNVFSLKGDRWREMRNTLSPSFTANKMRFMFDLVSKCSHDFVSCLHDRLESSSSEIEGKNLFTRYSNDVIATVAFGISVNSIEHPDNEFYRRGIDVSTFSGTFRFIKFMLFRLNPRLTRMAGFTFLSRATSKFFWRVISETVTARKRRGIVRPDMIHLLMQATDSKKKSIHQTMTIDDIVAQAFIFFLAGFDTTSTLMCYVVHELALHQDVQRRLREEVDRVLDDGTEISYEDTLGMEYLEMVISETLRMHPPTLLIDRQCAKEFHLPPAGPGYESVTIHPGENIWFPVLAIHRDPAHFPDPDKFDPERFNRENRNGIDPYTYIPFGVGPRKCIGNRFALMETKLLIIRLLRKFVIKPCERTMDPIVYKKGNFTLMPKDGFWVTFEKRNDH*

[123] [124]

>CYP9P2 name changed from CYP335A2 Am3_14.8 (443001-444560), 519 aa, 1 exon

version 2 = CYP335A2 Am2_14.6 (443001-444560), 519 aa, 1 exon

version 1.2 = CYP335A2 Am1.2_14.13c (15290-16849) plus, 519 aa, 1 exon

version 1.1 = CYP335A2 Am1.1_14.7b (395285-396839) minus, 519 aa, 1 exon

39% to 9f2m 63% to 9P1 (335A1) 40% to 9K1 51% to 9E2 partial, 54% to 9J7 partial

MESPTLLFSFELLAIGLTAIVLAKFVSLLHHQYNYWRKRRVPHVGAVPVLGSSWRIFTRRMSLPNFCSLVYKHRPGSRYLGMMDCFTPVVVVRDPNLIKEIAVKNFDHFPDHHSFINEKIDPIFGKNVFSLKGDRWREMRNTLSPSFTASKMRFMFDLVSNCSEEFVRYLYDHPEFSSSIEAKDAFTRYTNDVIATVAFGISVNSMENRDNEFYTKGADATNFGGIFRLFKFMLFRVNPRLTRMAGLSFLSRGTATFFHRVVRETVRARDERRIVRPDMIHLLMQARDKEDRRPVATVDNRMTIDDITAQAFIFFLAGFDTSSTLMCYVAHELALNPPVQERLREEVDRFMDGGNGAITYEALLKMEYMDMVTSETLRKYPPIVFIDRLCVEKFELPPAEQGYDHLIVHPDNIVWFPVYGLHHDPKYFPEPEKFDPERFNDANKRNIVPYTYMPFGLGPRKCIGNRFALMETKILIAYMLRKFRIKRTEKTRASIEFSKTNFSLTPDHGFWIGLEKRDP*

>Am2_14.6 Am3_14.8 (441001-442329), 426 aa, 1 exon, added to both N- and C- terms

version 2 = Am2_14.6 (440001-441185), 379 aa, 1 exon, psuedogene like 335A2

new in version 2.0

MEILSTSSVFTITVIALITVILSNFFPADTRIGRTIITWLASDSFRYSAHFANSSS*KNPVGPVLAQSSSKREILLDFTLNPSINQGDSFDRFIDRSSTRNIFSF*GAKGRKR*GILSPSFTMNEIDVRNCSQVCGSFALIYPRWTQRKILSSATT*SFIYQILLESV*IRWKTTSQFPLMRFRVHD**DY*E*FLKSTIFDFVPRVVSNGIEWRTRHR*TGYTAFIARNKEKSSIDKMTIDDIVSRAIRFSTWMALTLALSNFMRFVLYDPAFGWRGKEIDG*NILRILRRVYTGMMMSETLGKYSGLIFVDRVCTRKFALSGAGYDGATLYPGDIVFSPYVGSRS*IFFRSRNEKNEGNIVPYWNLLLEIKIFMIPSRRFSIRLNEKTPEAYRLSEK*SCIHS*RRNLD*IGRKERL*ILWLVKNFVHYFAKRFMILMKV*

>CYP9Q1 name changed from CYP335B1 Am3_14.8 (526001-527533), 510 aa, 1 exon

version 2 = CYP335B1 Am2_14.6 (530001-531533), 510 aa, 1 exon

version 1.2 = CYP335B1 Am1.2_14.13h (106877-108409) plus, 510 aa, 1 exon

version 1.1 = CYP335B1 Am1.1_14.7c (305231-306757) minus, 510 aa, 1 exon

39% to 9E2 35% to 9f2m 57% to 9Q2 (335B2) 42% to 9P1 (335A1)

MDYLQLGLTLLAILVAVYYLSTRNHKLLKRHGIVHIPPTPLFGNLGPLVRRKCHMEDVIQRVYDLDPDARYVGMYEFTTPLIIIRDPELIKTIGVKEITNFTNHRPFVDVGVDPMLGEVLFAMQGDRWREHRTMLTTLFTSSKIKSMFVLMSDCAKRFADYLSKVEREIELKSVLTRYTNDVIARCVYGVSVDSVNEPENIFYRYGQVASQLSTFKQNLMIFVHRNSPRLARLFNLKILPVHIEKFFHRLVMDTIETRRREGVHGLDMLQQLMDMQSRRKESEEGKRGMTVTDIANHAFSFFFGSVDTMATQISLISHMLAVNPDVQQRLQEEIDEVLSASEDKQVGYDVIQEMKYLDAVMSEAMRYHPILLFVDRVCGETFELPPALPGARPFKLERGMNIWFPVKAIHHDPKYFENPDRFDPDRFLRDGKGIASSGAYMPFGMGPRKCIGSRFALTEMKILLFNILAKCSFKVGSKTMVPLKFKEGVFNPVAKNGFWLKIERRENSCC*

[125] [126]

>CYP9Q2 name changed from CYP335B2 Am3_14.8 (599999-598401), 532 aa, 1 exon

version 2 = CYP335B2 Am2_14.6 (589998-588400), 532 aa, 1 exon

version 1.2 = CYP335B2 Am1.2_14.13g (159215-160813) minus, 532 aa, 1 exon

version 1.1 = CYP335B2 Am1.1_14.7d (252241-253839) plus, 532 aa, 1 exon

57% to 9Q1 (335B1)

MEFLSLALVLAAISIIAYYYCFVRKNFNLFQEHGILHVPPSPLVGNFGPLIRGKENVHDTIQRIYNIHPDAKYVGIFEFLTPVIMIRDLDLIKSITMKNFDQFPDHRPMFCKSVDPMLGEMLFIMDGERWKEHRNMLSPTFTSSKIKTMFVHMSECAKRFAHHLSKLPEKDRETEMKALLTRYTNDVIAACIYGVNVDSIKEPRNVFYMYGRVGATLIGLKKNLKIMVHRNMPWLANLLRLNILERHIAKFFTDLVVETVEERERNGTTNSDLIQLMMDTRNKKESGKKNLTVQNMANHAFSFFFGGFDTVSSQTCVLLHMLVENPEVQQRLQQEIDETLESNNGQLSYDVIQEMRYLDAVINEILRLHPIAVFIDRMCVKSFELPPALPGDVPFTVKPGMNVWIPVKAIHHDPRYYDEPEKFKPERFLDNGKNIIGSGAYFPFGIGPRICIGNRFALIEMKVLVCHILAVCDIKAGARTGIPLEFEKGVFNATAKTGFWLKIEPRKYSYHSGQINGLVNNHVINGACKTGI*

[127] [128]

>57889394 Am3_57889394 (4-286), 95 aa, 1 exon

version 2 = Am2_57889394 (4-286), 95 aa, 1 exon, 335B2 fragment, 2aa changes, probably sequenced from the other haplotype

ETVEDTERNGTTNSDLIQLMMDTRNKKESGKKNLTVQNMANHAFSFFFGGFDTVSSQTCVLLHMLVENPEVQQRLQQEIDETLESMMNYSKYQFS(1)

[129] [130]

>CYP9Q3 name changed from CYP335B3 Am3_14.8 (585999-584446), 517 aa, 1 exon

version 2 = CYP335B3 Am2_14.6 (574998-573445), 517 aa, 1 exon

version 1.2 = CYP335B3 Am1.2_14.13d (145027-146580) minus, 517 aa, 1 exon

version 1.1 = CYP335B3 Am1.1_14.7e (266474-268027) plus, 518 aa, 1 exon

58% to 9Q2 (335B2) ESTs BI508364 BI516506 BI505081 BI505012

MDYLTISLSLITVFVAVYYLATRNNDFFKKHGIPHVPPVPFLGNMGSLVRQKSNLHDVIDRTYNLDPGAKYVGIYEFTTPIIILRDLDLIKTITMKYLDHFPDHRSFAYEGADPVFGSMLFAMKGERWKEHRNMLTPTLTSSKIKGMFKLMTECAVRFADFLSVLPENERETEMKALLSRYANDVIASCVYGVSVDSINDPKNIFYVYGRRGTNVVGLKKSMFVLIHRNMPWLAKLFGLRFLEKHVQKFFYDLVYETIESREKLGTNRSDVLQLLMDIRDKANSSGKMTTMTVENVAIHAFTFFFGGFDSITSVTTLLTQMLAEHPDVQARLQQEIDETLRSNDGVLTYDAVHGMKYMDAVINETMRFCPVLPFLDRMCVESFQLPAPVPGGQPFTLRPGMNVWIPLAAIGRDPEYFEDPDKFDPDRFLNPEAGIKNSGAHFPFGLGQRKCIGERFAMMEMKVLLCYVLAACNVRIGSKTTVPMKLEKGLINANVKGGFWLKIEPRKVTYYNSSRSN*

[131] [132] [133]

>CYP9R1 name changed from CYP335C1 Am3_14.8 (580001-581596), 531 aa, 1 exon

version 2 = CYP335C1 Am2_14.6 (581001-582596), 531 aa, 1 exon

version 1.2 = CYP335C1 Am1.2_14.13f (153220-154824) plus, 531 aa, 1 exon

version 1.1 = CYP335C1 Am1.1_14.7f (258230-259825) minus, 531 aa, 1 exon

44% to AmGroup14.7d 49% to AmGroup14.7g

MLDSWSITAAIVAVLAIAYYQLIWKYKHFERIGIPCYHSIPLLGSFWEAVIQRNNFAEISRKIYNSYPDTKYMGMYDTTTPVLLIRDTELIKAISVKHFEQFPDHRSFQNEATDPLFAKNLFALRGDRWREIRNLLSPAFTSSKMKSMFILMRDCAKEYGDYFASLTGDESTIELKDAFTRYTNDVIATCAFGVEVNSMKDRKNKFYVYGREGTTFGSWASIKFFVTRVLPVSVCTLLRIRLIRKEISDFFIDLVSTTIKTREEKGIVRPDMIQLMMESKGKLGAGKEMSMIDICAQAFVFFFGGFESTSTLMCFAAYEIAVNEDIQRRLQNEIDQVLEERDGEVTYAAVNEMKFLDAIIYEALRMYPVVVATDRVCMKPFELPPNRPGEKPYLLKEGDNVWFPIYAIQRDPQYYPEPDKFDPDRFLNDTKQMINSGLFLTFGIGPRMCIGNRFAMLETKVLLFHLFARCNLVPCSKTTIPMKLNRKGFSMTAENGFWFKIEPRSAKKEEKIAVPGTTMLIDKIPDRYPDN*

[134] [135]

>CYP9S1 name changed from CYP335D1 Am3_14.8 (572001-573524), 507 aa, 1 exon

version 2 = CYP335D1 Am2_14.6 (577001-578524), 507 aa, 1 exon, 1 aa change

version 1.2 = CYP335D1 Am1.2_14.13e (151195-149678) minus, 507 aa, 1 exon

version 1.1 = CYP335D1 Am1.1_14.7g (261868-263392) minus, 507 aa, 1 exon

49% to 9R1 (335C1)

MAIFALLLIVLGILGSYHLLKSQNPFKEHGLPYKSYLPILGSTWESILRRKSFAVVIQEIYNLAPSARYVGFYNRTTPIVMIRDPELIKTIAVKNFDAFRNHRTVNDTQTDDVLLSGNLLLLRDNRWREVRSLNTPAFSTSKIRSMYRSMSEIAINVARYLSTLAPGQNIVEMKDIFTRYANDVFATCAFGISVDSLSDRENKFYELGREALDIHSTPILKLILIFAFPKLARRLGVSLVSKEATNFFTRVVSENIKMREEKGITRPDFIQAMIDKRNGRGRDDELTVEDITAQAFVFFFGGFETTSGLLSFAVHELAANPEIQGKVHAEIDRVLVSNNEITFERVNGLMYLDAVINETLRMYPIIPITDRECSKRFELPPVLPDAKPYVLKEGSHVWFPIYAIQRDPRYFEKPDCFDPDRFLDDNKKRSDAFNGDAYMPFGAGPRNCIGNRFSMVETKVALFHILAKCRLDVCPKTTIPMELRKRGVFLTAKNGFWLRIVPRHPVT*

[136] [137]

>CYP336A1 Am3_2.7 (983001-984491), 496 aa, 1 exon

version 2 = CYP336A1 Am2_2.8 (230001-231491), 496 aa, 1 exon, 5aa changed

version 1.2 = CYP336A1 Am1.2_Group2.13 (199618-201108) plus, 496 aa, 1 exon

version 1.1 = CYP336A1 Am1.Am1.2.15 (200938-202428) plus, 496 aa, 1 exon

34% to 6AH1 Anopheles EST BI946448 from brain

MASAFLTLVTGALLLLCFYLYLKYTYWKRNGIPYSKGYYPIIGHFLPLIMKKQSYSEIIEEIYRDSNHSMVGMYKGMKPVLILRDINLIKTVLQSNFSKFHENAVKIDPKLDPLLAKNPFFCYGELWQTGRKRLTYAFSNARLKILFAAVYEVCTKFRNFLDRRLESSKKYEVELKSLFLKFTSEVVANAGLGIEGFCFEDDKVQSMFTNLDNNDFLDTFLIGIIVHFPFLTKLLRIQFLPTKHDKFFRTVVKKNLELRKSDPIPRNDFIQLMIEMEQTGEKIDEEIVAAHAVSFYLDGVETSSVTLNFIGCQLAIHQDVQEKLRKEVRSTLEKHGGVLTFEALKDMTYMNQVISESQRYFSALGFLGKICTDEFELQGSDGLNYRAKPGTELLIPICGLHKDPKYWDNPEIFDPERFSDENKQRIEKMAFIPFGEGPRICVGMRMAMLQMKSCLATLMKDYKLEVSPKMQLPLKLSPTYFLSAPLGGGWVLISKA*

[138] [139]

CYP4 clan (4 seqs, 3 complete, 1 incomplete)

[140] [141]

>CYP4G11 Am3_16.8 (1445459-1449343), 545 aa, 7 exons

version 2 = CYP4G11 Am2_16.8 (1440635-1444519), 545 aa, 7 exons

version 1.2 = CYP4G11 Am1.2_Un.3078 (673-4557) minus, 545 aa, 7 exons

version 1.1 = Am1.1_Un.2145 (673-4743) minus, 545 aa, 7 exons

CYP4G11 63% to 4g15m, 100% to AF207948 partial seq

MAAASATGFSASSVFLSLLIPALILYFIYFRISRRHLLELAEKIPGPPALPLIGNALDLFGT (1)

MFSQVLKKAENFKDVVKIWVGPKLVICLIDPRDVEIILSSNVYIDKSTEYRFFKPWLGDGLLIST (1)

GQKWRNHRKLIAPTFHLNVLKSFIDLFNANARSVVEKMRKENG

KEFDCHNYMSELTVDILLETAMGVSKPTRDHNAFEYAMAVMK (2)

MCDILHLRHTKIWLRPDWLFNLTKYGKNQIKLLEIIHGLTKKVIQLKKEEYKSGKRNIIDNSAQKTESK (0)

TNNIVVEGVSFGQSVGLKDDLDIDDDVGEKKRQAFLDLLIEAGQNGVLLTDKEVKEQVDTIMFE (0)

GHDTTASGSSFFLAVMGCHPDIQEKVIQELDEIFGDSDRPATFQDTLEMKYLERCLLETLRMYPPVPLIAREIKTDLKLA (1)

SGDYTIPAGCTVVIGTFKLHRQPHIYPNPDVFDPDNFLPEKTANRHYYAFV PFSAGPRSCVGRKYAMLKLKIVLSTILRNFRVRSDVKESEFRLQADIILKRADGFKIRLEPRKQVASTA*

[142] [143]

>CYP4AV1 Am3_Un.279 (37180-27401), 501 aa, 8 exons

version 2 = CYP4AV1 Am2_Un.282 (45375-55154), 501 aa, 8 exons, insertion of nucleotide at C-end of 1st exon changing 5 aa and donor splice site

version 1.2 = CYP4AV1 Am1.2_Un.1387 (15087-18877) plus, 124 aa, 2 exons

version 1.2 = Am1.2_Un.1625 (113-5079) plus, 437 aa, 7 exons

version 1.1 = CYP4AV1 Am1.1_Un.2000 incomplete on N-terminus (1-4856) plus, 496 aa 7 exons

39% to 4c3

MSSSTIIMGVSWLTMILSICLMTIVVLLLVRRGKFLYALRKVPCPPAFPIIGNAYELCCSPEE(1)AFKKMIKWGKELGDMYLIWVGMRPFIFLYKAEAIQPLLSSSVHIDKSLEYQYLQPWLGSGLVTSTG(1)EKWHFHRKLLTPTFHSGLLELYLKTTIREAQILISCLRKEIGKPEFDIVPYAKRAALDIICD(1)SSMGCNINAQKNFENEYVQAVNT(2)LASISQRRFLNVWMSFDPIFKLTSWGKRHDHALSVTHGFVNK(0)IIAERKAEWKDRKDTNFNEKSHKRQALLDLLLELSKDGKVLTDDDIRDEVNTFMFAGHDTTATSVSWILYALGRHPQYQ(0)ELIIEEYDETVGTKELTLDILSKLTWLEACIKESWRLYPVTPLIARQIYHPITIL(1)GHEIPIGSTVLVNSFLLHRDSRYFPEPDIYRPERFLPDGPKYPSYAFVPFSAGSRNCIGWKYGTMIVKVLILYILKNFHVESLDTEDQLRFISELVLHNADGLRLKITPRK*

[144] [145]

>CYP4AZ1 Am3_Un.3788 (1270-4633), 513 aa, 12 exons, one amino acid change with correction of intron-spanning codon

MISAILFFIFLLATLHYFLLHHRKFGKMINLIPGPEPLPILGNIPTFHNISPS(1)ELWKFLTQLSKQYYPIYRMWTFLEAYVHICHPDDIE(0)TILGNIKFTKKGFGYKYLKPWFNTGLLTSSG(1)HKWHVRRKILTSAFHFNVLRQFVDIFIEDAERLIKTLESEEGIFVENLLQLTSEHTLNVICE(1)TAMGTSLKNKEKFQYEYRKAVYNMGCIFANR(2)IVKPWFYYDFFFNLSPEGWQQSKLLKILHNFTRK(0)IIQERKEYHDKTNGRYLNDFHENINENDNNNDYNDF(1)RRKRLAMLDLLIEAHRNNKIDDEGIREEVDTFMFR(0)GHDTTAISFCFSIMLLAEHKEIQ(0)DRARAEIKAAIEENGGKLNITVLQNLPYLERCIKESLRLFPSVPRISRKLETSVKLS(1)NYEIPSNTIINVNIFDTHRDPKFWPNPNKFDPDRFLPENSKKRHPYAYVPFSAGPRNCIG(1)QRFAMLELKTYLGLLLYNYYFEPIDYLKDVTFVSGIVLRLENPVRMKFIPVKKIC*

version 2 = CYP4AZ1 Un.1527 Am2_Un.5748 (1270-4633), 487 aa, 12 exons, added 7th intron, completed 11th exon

version 1.2 = Am1.2_Un.1527 (1556-4363) plus, 478 aa, 10 exons

version 1.1 = Am1.1_Un.8281 incomplete on both ends (644-1499) plus, 132 aa, 3 exons

version 1.1 = Am1.1_Un.2540 incomplete on N-terminus (5369-5938) minus, 71 aa, 2 exons

50% to 4AB2 fire ant complete seq, revised to match fire ant seq

MISAILFFIFLLATLHYFLLHHRKFGKMINLIPGPEPLPILGNIPTFHNISPS(1)

ELWKFLTQLSKQYYPIYRMWTFLEAYVHICHPDDIE(0)

TILGNIKFTKKGFGYKYLKPWFNTGLLTSSG(1)

HKWHVRRKILTSAFHFNVLRQFVDIFIEDAERLIKTLESEEGIFVENLLQLTSEHTLNVICE(1)

TAMGTSLKNKEKFQYEYRKAVYNMGCIFANR(2)

IVKPWFYYDFFFNLSPEGWQQSKLLKILHNFTRK(0)

IIQERKEYHDKTNGRYLNDFHENINENDNNNDYND (1)

[fire ant seq. QYLKNFNQSIITDNEEIVGS]]]

[fire ant seq. QYLKNLNKDVVPNETETIGI]]]

VRRKRLAMLDLLIEAHRNNKIDDEGIREEVDTFMFR(0)

GHDTTAISFCFSIMLLAEHKEIQ(0)

DRARAEIKAAIEENGGKLNITVLQNLPYLERCIKESLRLFPSVPRISRKLETSVKL(1)

[[[fire ant seq. ISRITTEEAQL

[[[fire ant seq. ISRITSEETEL

RNYEIPSNTIINVNIFDTHRDPKFWPNPNKFDPDRFLPENSKKRHPYAYVPFSAGPRNCIG(1)

KSHLIP]]]

KTYLIP]]]

QRFAMLELKTYLGLLLYNYYFEPIDYLKDVTFVSGIVLRLENPVRMKFIPVKKIC*

[146] [147]

>CYP4AZ1-frag Am3_49183273 (191-59), 45 aa, 1 exon

(0)DRPPAEIKAAIEENGGKLNITVLQFFSDLERCIKESLRLFPTVAG(1)

version 2 = CYP4AZ fragment Am2_49183273 new (163-31), 56 aa, 1 exon 79% to Un.1527 in 4 clan

(0)DRPPAEIKAAIEENGGKLNITVLQFFSDLERCIKESLRLFPTVAG frameshift

GSKLETSRVKF (0?)

[148] [149]

>CYP4BA1 Am3_Un.763 (16669-15582), 298 aa, 3 exons, missing N-term, changed intron acceptor on N-term to remove one aa

version 2 = CYP4BA1 Un.1327 Am2_Un.1509 (16763-15674), 299 aa, 3 exons

version 1.2 = Am1.2_Un.1327 incomplete on N-terminus (901-1990) plus, 299 aa, 3 exons

version 1.1 = Am1.1_Un.3324 incomplete on N-terminus (901-1990) plus, 285 aa, 3 exons

52% to 4aa1 55% to 4AN3 partial

(1)QIMLLYRMIRPWLLIEWIYRLTKYGREEEKQRKNLFDTCFKMVKEKRDLLQSKDRISNNDIKKNK

NISLLEYMVEINEKNPCFSDEDIVEECCTFMLAGQDSVGTATAMTIFLLANHPEWQNKCIEEIDEIFNG

DTRFPTISDLKEMKCLEMCIKESLRLYPSVPIIGRTLGEDIKI (1)

GKHIIPAGCSVLISPYSTHHLPHHFPDPDTFKPERFNSENSEKRHPYAYIPFSAGPRNCI (1)

GYKFAMLEMKSIISAILRKCRLQSIPGKKEIRPKFRMTIRAQGGLWVKIIERDQILKSIAA*

[150] [151]

mito clan (6 sequences, 6 complete)

[152] [153]

>CYP314A1 Am3_5.26 (289219-286399), 517 aa, 10 exons, annealing of PCR primers I've designed support first exon, which was a guess

version 2 = CYP314A1 Am2_5.28 (288891-286071), 517 aa, 10 exons (first intron is a guess)

version 1.2 = CYP314A1 Am1.2_5.28 (13954-16400) plus, 516 aa, 10 exons (first exon is a guess)

version 1.1 = CYP314A1 Am1.1_5.25 AADG02002903.1 incomplete on N-terminus (17670-19852) minus, 471 aa, 9

exons (19852-19708, 19574-19378, 19298-19111, 19031-18829,

18707-18579, 18471-18305, 18238-18160, 18072-17875, 17777-17670)

43% to 314A1, 55% to pea aphid (Acyrthosiphon pisum) P450 below

cyan is extra seq added based on CF588143

note This seq is less than 55% identical to Drosophila and Anopheles 314A1s,

but there appears to be only one orthologous seq in each species and it does not make

sense to give the ortholog a different subfamily.

MLLSSAWFEVIAAVLLTILIFVTSHRPAWWFWTATSHEASA(1)PAEGKFKTVSKVPGPFSLPIFGTRWIFSCIGYYKLNKIHDAYKD(1)LNQRYGALCKEEALWNFPMISVFSRQDIETIIRRNSRYPLRPPQEVISHYRRTRRDRYTNLGLVNE(2)QGQTWHDLRVALTSELTAASTVLGFFPALNIVADSFIELIRRQRVGYKVTGFEELAYKMGLES(1)TCTLILGRHLGFLKPDSSSELATRLAEAVRIHFTASRDAFYGLPLWKLLPTCAYKQLIESEDAIYN(2)IISEIIETTIQEKRDDAKDESVEAIFQSILRQKNLDIRDKKAAIVDFIAAGIHT(0)LGNTLVFLFDLIGRNPAVQNKLYEETYALAPAGCDLTIDNLRKAKYLRACITESLR(2)LIPTTTCIARILDEPIELSGYRLTAG(0)TVVLLHTWIAGLNEENFKDAKKYLPERWTTPTTPHSPLLVAPFGAGRRICPGKRFVDLALQLILAK(0)IIREFEIIVEEELDLQFEFILAPKGPVSLGFRDRS*

[154] [155] [156]

>Am2_59135075 Am3_59135075 (103-32), 24 aa, 1 exon, similar to CYP314A1

version 2 = Am2_59135075 (72-1), 24 aa, 1 exon, like 314A1, but with 2aa changes, probably from the other haplotype

(0)LGNTLVCLNDLIGRNPAVQNKLYE(0)

[157] [158]

>CB336480 Tribolium castaneum embryonic cDNA

MFEKIFQSLDVTSLLIIAI

FFLFLEYRPPWWYRNNDCKKGVKLIPGPLALPGLGTTWIFFFGGFSFNRLHLYYENMYKR

YGPVMKEEYWCNIPVINLFEKREIVKVLKAGGKYPLRPPVEAVAHYRRSRLIDTLALG

[159] [160]

>CF588143 pea aphid (Acyrthosiphon pisum) 48% to 314A1 D. pseudoobscura

2 RVLRQSGKYPIRPPNEVTANYRKSRPDRYTNTGLVNEQGEVWAMLRNKLTPELTSPRTIR 181

182 RFLPEVNQLADDFNNLISLARDGNNVVRGFEGYCNRMGLES 304

305 TCTLILGRRIGFLDGEVSETATRLADSVTSQFRASQEAFYGLPLWKLIPTKAYKDFVAS 481

482 EDALYDIVSEFVESALIDEQQSFTDVRSVFVSILQASELDNRDKKAAIIDYIAAGIKT 655

LGNTLVFIL 682

[161] [162]

>CYP334A1 Am3_9.4 (739428-736819), 531 aa, 10 exons

version 2 = CYP334A1 Am2_9.4 (736455-733846), 531 aa, 10 exons, second exon is a guess

version 1.2 = CYP334A1 Am1.2_9.8 (72740-75349) plus, 530 aa, 10 exons (second exon is a guess)

version 1.1 = CYP334A1 Am1.1_9.6 alternate splicing possible? (1158493-1161102) minus,

514 aa, 9 exons (1158603-1158493, 1158868-1158668, 1159034-1158959,

1159285-1159117, 1159526-1159346, 1159843-1159593, 1160068-1159927,

1160289-1160153, 1161102-1160830)

34% to 12A5m a new family in the mito clan

MTESQTASVDESLRTDTIPLLDHTATTEVSPTTFEVSTMKVDTQIFDKAPLPFDEIPGPAILKIWEKYWKYVPLLG(1)

RLTWNRNITPLKYLFNEYGCIVRINGPLSGDIVMIHR(2)

PEHIAEVFKQEGDTPVRSGIDILQHYRLNYRKYRLAGPFSM(2)

QGTEWLEIRDKVEDTFNQISSTFFTKIDTCCNELITRICKIRNRQNE(0)

VPVSFYEDLIRWAMECFCDLTFNKRLGFLEPIGYNSSSEASKLINALTTAHKYM

SRCETGFQVWRFFLTPFARKLFEACDVLDN(2)

VIGKYVRQAQCKLRIRKSHSEESSMTERSPVLEKLLLNEGIHPDDICTMLMDMIILGIQA(0)

TVNSEAFLLYHLAKNPRTQRKVYDEIISVLSNDNSSFTEKSLKNMPYLKACIQETLR(2)

LHPAIPYITRLLPKTISLHGYTIPKG(0)

TFVIMANQITSQREENFEDPFKFWPERWLSNSSKEDVHFSYLPFGHGIRSCLGKNMAEAKMMLLTAK(0)

LVRQFRIEYDYADIKSRFMMVNVPNKPLRFRFVNRN*

[163] [164]

>CYP315A1 Am3_Un.436 (31787-34039), 535 aa, 6 exons

version 2 = CYP315A1 Am2_Un.248 (31787-34039), 535 aa, 6 exons

version 1.2 = CYP315A1 Am1.2_Un.768 (10162-7910) minus, 535 aa, 6 exons

version 1.1 = CYP315A1 Am1.1_Un.1189 (10257-12510) minus, 535 aa, 6 exons

38% to 315a1m, probable mito clan, maybe 315Bx

note This seq is less than 55% identical to drosophila and anopheles 315A1s,

but there appears to be only one orthologous seq in each species and it does not make

sense to give the only ortholog a different subfamily.

MNLAQNILKSGKSVSLSSNVIALKYNVPGCGYAGASQTSRIDDLSDISKSTDGGNRSKIEITEKLRDRNYGTVAVATSESILQEMPEPRGIPVFGTLFSFILSGGPKKQHEYVDKRHKELGPVYKERIGPTTAVFVNSIHEFRK

IFRLEGSTPKHFLPEAWTLYNEIRKCRRGLLFM(2)

NGEEWVYFRKILNKVMLLPDPTNLMIAPCQEVAIELKRKWQKQIKTNNIISNLQVQLYQWSIEA(1)

MMATLMGSYWYSYKHQLSRDFEILAETLHEIFEYSAKLSIIPVKLAMNLRLPVWKKFVASA

DTAFEIVRMLVPEMAKLGGNGLLKKMMDEGIRAEDAICIVTDFILAAGDT(0)

TATTLQWILLLLCNHPEKQEELFKHLKDLSQEDILRLPLLKGIIKESLRLYPIAPFISRYLPEDSVIGNYFVPKG(0)

ELLVLSLYSSGRDAANFPQPNEFRPERWIRTQKGIYQGVVHPHASLPFALGARSCIGRKLAEIQISFALAE(0)

LIKSFKIECINKNQVKLILHLISVPSQSIKLKLMERN*

[165] [166]

>CYP302A1N Am3_Un.5946 (2916-5374), 228 aa, 4 exons, N-terminal part of CYP302A1

version 2 = CYP302A1N Am2_Un.4726 (2916-5374), 228 aa, 4 exons, added to C-terminus

version 1.2 = CYP302A1 N-term Am1.2_Un.8443 incomplete on C-terminus (289-2450) minus, 202 aa, 3 exons

version 1.1 = CYP302A1 N-term Am1.1_Un.2216 incomplete on C-terminus (289-2832) minus, 202 aa, 3 exons

45% to CYP302A1 aa19-183 in CYP302 family, mito clan (disembodied)

MCTLLKKCNQSIRKKLFIKFYSNEFTKSKIKINHSQPKAFYDIPGPKSLPIIGTLYKYLPFIG(1)

EYSFTNLYESGKKKLKCFGPLVREEIIPNVNVIWIYRPEDIAEIFKAESGLHPERRSHLALLKYRKDRPNIYNTGGLLPT(2)

NGSEWWRLRKEFQKVSSKPQDVINYLKETDCVIQEFVELCNNEKFADFLPLLSRLFLEL(1)

TCLVVFDIRLNSFSKEERCENSISSK

[167] [168]

>CYP302A1C Am3_Un.9312 (7517-4855), 295 aa, 5 exons, C-terminal part of CYP302A1

version 2 = CYP302A1C Am2_Un.1798 (7517-4855), 295 aa, 5 exons, added to N-terminus

version 1.2 = CYP302A1 C-term Am1.2_Un.2118 incomplete on N-terminus (3101-5209) minus, 250 aa, 4 exons

version 1.1 = CYP302A1 C-term Am1.1_Un.3145 incomplete on N-terminus (122-2230) plus, 250 aa, 4 exons

51% to 302A1 (disembodied)

note these two pieces are probably from a single gene missing about 70aa inbetween

SSKLIKAAFATNSAILKLDNGLQLWRLFETPLYRKLRKAQTYMEM(2)

IALELVSRKKNNMKIRYNKSFLDAYLENPVLDIKDIVGMACDMLLAGIDT(0)

TSYSTAYILYHLAKNQNIQEKLRIEATQLLKNHNEPISINILRNASYTKAVIKESLRLNPISIGIGRILQTDVVLSGYRVPKG(0)

SVVVTQNQIICRLPEYFEEPNLFIPERWLREYSENNNKINYKKTVHPYVLLPFGHGPRSCIARRFAEQNMQILLLR(0)

ICRRLKISWHGDDLGMISLLINKPNALLKFNFHDILNNNSV*

[169] [170]

>CYP302A1 (Am3_Un.5946, 225 aa and Am2_Un.9312, 295 aa), combined seqs 520aa, 7 exons, only 3 aa overlap between CYP302A1N and CYP302A1C, 50% to 302A1 Dm

MCTLLKKCNQSIRKKLFIKFYSNEFTKSKIKINHSQPKAFYDIPGPKSLPIIGTLYKYLPFIG(1)

EYSFTNLYESGKKKLKCFGPLVREEIIPNVNVIWIYRPEDIAEIFKAESGLHPERRSHLALLKYRKDRPNIYNTGGLLPT(2)

NGSEWWRLRKEFQKVSSKPQDVINYLKETDCVIQEFVELCNNEKFADFLPLLSRLFLEL(1)

TCLVVFDIRLNSFSKEERCENSISSKLIKAAFATNSAILKLDNGLQLWRLFETPLYRKLRKAQTYMEM(2)

IALELVSRKKNNMKIRYNKSFLDAYLENPVLDIKDIVGMACDMLLAGIDT(0)

TSYSTAYILYHLAKNQNIQEKLRIEATQLLKNHNEPISINILRNASYTKAVIKESLRLNPISIGIGRILQTDVVLSGYRVPKG(0)

SVVVTQNQIICRLPEYFEEPNLFIPERWLREYSENNNKINYKKTVHPYVLLPFGHGPRSCIARRFAEQNMQILLLR(0)

ICRRLKISWHGDDLGMISLLINKPNALLKFNFHDILNNNSV*

[171] [172]

>Am2_98065759 Am3_98065759 (650-5), 72 aa, 2 exons

version 2 = Am2_98065759 new (612-33), 72 aa, 2 exons 62% to 302A1

(0)IIALHLWRFLAPPLYRKLRNAQTYMET(9)NWYLVTPNHMKIRSHKSFLHPYLENPALHIKAILGMACVMNYLSL

[173] [174]

>CYP49A1 ortholog Am3_Un.193 (102533-109096), 531 aa, 10 exons

version 2 = CYP49A1 ortholog Am2_Un.86 (102533-109096), 531 aa, 10 exons

version 1.2 = CYP49A1 ortholog Am1.2_Un.343b (13080-19643) minus, 531 aa, 10 exons

version 1.1 = CYP49A1 ortholog Am1.1_Un.423a incomplete on N-terminus and C-terminus of exon 6

(214-3539) plus, 312 aa, 7 exons

AmGroupUn.10899 incomplete on both ends (1221-1394) minus, 58 aa, 1 exon

46% to 49A1, 50% to 301A1 mito clan Combined Am1.1_Un.423a and Am1.1_Un.10899,

MLKIFVIKFLCCRVKMQILNCKFTKLLKNTVKQSNNIIKTLETLTTEVEEQDWSRCRPYSEIPGPKPIPFLGNTWRFIPFIG(1)

DFKIQAVDQVSKKLYKEFGDIVKVEGLLGRPDMVFIYDANEIERIFRQEERMPYRPSMPSLNYYKHVLRKEFFKENAGVIAV(2)

HGESWYNFRSKVQQVMLQPRTARMYITSMEEASLAFLER(2)

IKKIRNKNDEVPDDFLNEIHKWSLES(1)

IARVALDVRLGCLDDDANIETQQLIDAVTTFFKNVGILELKIPFWKLFNTPTWLKYVNALDTILS(2)

ITSRYTTVALSRTKEAEKSDKEPSLLERVLALENDTKLATILSLDLFLVGIDT(0)

TSSTVASTLYQLALHPDEQDRAYNEVCNILPSKDMQLDGKHLDKLKYLKACIKETLR(2)

MYPVVIGNGRCMTKDTIIKGYRVPKG(0)

VQVVFQHYVISNLDKYFPHSDKFLPERWLQSDGVRHSFASLPFGYGRRMCLGRRFAELEMLVVISK(0)

ILQRYKIEYHHEKLEYYINPMYTPKGSLNLKFIDR*

[175] [176]

>CYP301A1 Am3_Un.193 (112558-109218), 516 aa, 10 exons, 1st exon is a guess

version 2 = CYP301A1 Am2_Un.86 (112558-109218) changed to better splice sites for 2nd intron (112558-109218), 516 aa, 10 exons

version 1.2 = CYP301A1 ortholog Am1.2_Un.343a (9993-12955) plus, 532 aa, 9 exons (exon 1 is a guess)

version 1.1 = CYP301A1 ortholog Am1.1_Un.423b incomplete on N-terminus and missing exon

(3686-6941) minus, 534 aa, 8 exons

69% to 301a1 D. melanogaster

MMTGKTRYLFFKIHFLLLYVLI(2)CMVDHDTTTIQQGKPYKDIPGPRPIPILGNTWRLFPMIGQYEISDIAKLSQIFYDEYGKIVRLTGLIGRPDLLFVYDVDEIEKIYRQEGPTPFRPSMPCLVHYKSVVRKDFFGSLPV(1)FTILRRHGEPWREFRTRVQKPILQPQTVRKYITPIEMVTSDFIQR(2)IQEIKGEDGEVPGDFDNEIHKWALEC(1)IGRVALDVRLGCLSSNLTSDSEPQKIIDAAKFALRNVAILELKAPYWRYVPTLLWSRYVRNMNYFIE(2)VCMKYIDATMERLKTKKAVDEYDLSLMERILAKETDPKIAYILALDLILVGIDT(0)ISMAVCSILYQLATRPEEQEKIYQELVEILPDPSVPLNMSHLDKAIYMKAFIREVFR(2)VYSTVIGNGRTLQNDTIICGYKVPKG(0)VQVVFPTVVTGNMEKYVTDAKIFKPMRWLKESTKTLHPFASLPYGHGARMCLGRRFADLEIQVLLAK(0)LIRSYKLEYHHKPLKYKVTFMYAPDGELKFKVLPR*

>44589653 Am3_44589653 (719-549), 56 aa, 1 exon, 47% to A. mellifera CYP301A1,

(2)GASLGNVTPG*QPEGILEGTRSNVMKVVILEINAHNWRNAPTLLWSRYVRHMNHFIE(2)

>Un.3179 Am3_Un.3179 (2-182), 61 aa, 1 exon, new, 52% to A. gambiae CYP303A1

FKTFSVLSKKYGPIIGMKIGINHVVVLNEYESIKSMLSNEDCDGRPFGILYDKRTGGINRG(1)

>49199747 Am3_49199747 (2-184), 59 aa, 1 exon, 93% to exon 1 of A. mellifera CYP6BE1

MFLTTWLIPDIIAVASLITVGLYFLYK*NLFKFWHKKGIFYVKTSFPTGNIMPIINGKFP*

>Am3_57847262 Am3_57847262 (819-522), 97 aa, 1 exon, exon 3 from CYP6AS subfamily; several in-frame stop codons and single nucleotide insertion at X, 55% to A. mellifera CYP6AS10

PLQSPQAFLFFVTGLKPRPKR*ATALFKLAQIHDIQDNVRDXEICRNIAYMLNGEWHYENITNMPYLEAVFKGIVHCY*HLDQTFPNLKRIKYSTLKSKS

[177] [178]