DHFR dihydrofolate

Alignment of vertebrate DHFR

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DHFR_homSap   MVGSLNCIVAVSQNMGIGKNGDLPWPPLR--NEFRYFQRMTTTSSVEGKQNLVIMGKKTWFSIPEKNRPLKGRINLVLSRELKEPPQGAHFLSRSLDDALKLTEQPELANKVDMVWIVGGSSVYKEAMNHPGHLKLFVTRIMQDFESDTFFPEIDLEKYKLLPEYPGVLSDVQEEKGIKYKFEVYEKND---
DHFR_panTro   MVGSLNCIVAVSQNMGIGKNGDLPWPPLR--NEFRYFQRMTTTSSVEGKQNLVIMGKKTWFSIPEKNRPLKGRINLVLSRELKEPPQGAHFLSRSLDDALKLTEQPELASKVDMVWIVGGSSVYKEAMNHPGHLKLFVTRIMQDFESDTFFPEIDLEKYKLLPEYPGVLSDVQEEKGIKYKFEVYEKND---
DHFR_gorGor   MVGSLNCIVAVSQNMGIGKNGVLPWPPLR--NEFRYFQRMTTTSSVEGKQNLVIMGKKTWFSIPEKNRPLKGRINLVLSRELKEPPQGAHFLSRSLDDALKLTEQPELANKVDMVWIVGGSSVYKEAMNHPGHLKLFVTRIMQDFESDTFFPEIDLEKYKLLPEYPGVLSDVQEEKGIKYKFEVYEKND---
DHFR_ponAbe   MVGSLNCIVAVSQNMGIGKNGDLPWPPLR--NEFRYFQRMTTTSSVEGKQNLVIMGKKTWFSIPEKNRPLKDRINLVLSRELKEPPQGAHFLSRSLDDALKLTEQPELANKVDMVWIVGGSSVYKEAMNHPGHLKLFVTRIMQDFESDTFFPEIDLEKYKLLPEYPGVLSDVQEEKGIKYKFEVYEKND---
DHFR_nomLeu   MVGSLNCIVAVSQNMGIGKNGDLPWPPLR--NEFRYFQRMTTTSSVEGKQNLVIMGKKTWFSIPEKNRPLKGRINLVLSRELKEPPQGAHFLSRSLDDALKLTEQPELANKVDMVWIVGGSSVYKEAMNYPGHLKLFVTRIMQDFESDTFFPEIDLEKYKLLPEFPGVLSDVQEEKGIKYKFEVYEKND---
DHFR_macMul   MVSSLNCIVAVSQNMGIGKNGDLPWPPLR--NEFRYFQRMTTTSSVEGKQNLVIMGKKTWFSIPEKNRPLKGRINLVLSRELKEPPQGAHFLARSLDDALKLTEQPELANKVDMIWIVGGSSVYKEAMNHPGHLKLFVTRIMQDFESDTFLPEIDLEKYKLLPEYPGVLSDVQEEKGIKYKFEVYEKND---
DHFR_papAnu   MARSLNCIVAVSQNMGIGKNGDLPWPPLR--SEFRYFQRMTTTSSVEGKQNLVIMGKKTWFSIPEKNRPLKGRINLVLSRELKEPPQGAHFLARSLDDALKLTEQPELANKVDMIWIVGGNSVYKEAMNHPGHLKLFVTRIMQDFESDTFLPEIDLEKYKLLPEYPGVLSDVQEEKGIKYKFEVYEKND---
DHFR_calJac   MVQLLNCIVAVSQNMGIGKNGELPWPPLR--NEFRYFQRMTTTSSVEDKQNLVIMGRKTWFSIPEKNRPLKDRINVVLSRELKEPPQGA-LLARSLDDALKLTEQPELANKVDMIWIIGGSSVYKEAMDHPGHLKLFVTRIMQDFESDTFFPEIDLEKYKLLPQYPGVPSDDQEEKGIKYKFEVYEKND---
DHFR_saiBol   MVQLLNCIVAVSQNMGIGKNGDLPWPPLR--NEFRYFQRMTTTSSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINVVLSKELKEPPQGAHFLARSLDDALKLTEQPELANKVDMIWIIGGSSVYKEAMDHPGHLKLFVTRIMQDFESDTFFPEIDLKKYKLLSEYPGVLSDVQEEKGIKYKFEVYEKND---
DHFR_tarSyr   MVRPLYCIVAVSQNMGIGKDGTLPWPPLR--NEFRYFQRMTTMSSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINLVLSRELKVPPPGAHFLAKSLDDALKLIEQPELANKVDKVWIVGGNSVYREAMSQPGHLKLFVTRIMQDFESDTFFPEIDLEKYKLLSEYPGVLSDVQEEKGIKYKFEVYEKND---
DHFR_otogar   MVRPLNCIVAVSQNMGIGKNGDLPWPPLR--NEFNYFQRMTKTSSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKESPQGAHFLAKSLDDALKLIEEPELANKVDMVWIVGGSSVYKEAMNQPGHVKLFVTRIMQDFECDTFFPEIDLKKYKLLSEYPGVLSDVQEEKGIKYKFEVYEKND---
DHF1_micMur   MVRTLNCIVAVSQNMGIGKNGDLPWPPLR--NEFRYFQKMTTTSSVEGKQNLVIMGRKTWFSIPEKSRPLKDRINVVLSRELKEPPQGAHFVAKSLDDALKLIEQPELANKVDMVWIVGGSAVYKEAMNQPGHLKLFVTRIMQDFESDTFFPEIDLKKYKLLPEYPGVLSDVQEEKGIKYKFEVYEKND---
DHFR_tupBel   MVRPLNCIVAVSQNMGIGKNGDLPWPPLR--NEFKYFQRMTTTSSVEGKQNIVIMGRKTWFSIPEKNRPLKDRINLVLSRELKEPPQGAHFLAKSLDDALKLIEQPELANQVDMVWIVGGSSVYREAMNQPGHLKLFVTRIMQDFESDTFFPEIDLEKYKLLPEYPGVLSDVQEEKGIKYKFEVYEKND---
DHFR_musMus   MVRPLNCIVAVSQDMGIGKNGDLPWPPLR--NEWKYFQRMTTTSSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQPELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEYPGVLSEVQEEKGIKYKFEVYEKKD---
DHFR_ratNor   MVRPLNCIVAVSQNMGIGKNGDLPWPLLR--NEFKYFQRMTTTSSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPQGAHFLAKSLDDALKLIEQPELASKVDMVWVVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLEKYKLLPEYPGVLSEIQEEKGIKYKFEVYEKKD---
DHFR_criGri   MVRPLNCIVAVSQNMGIGKNGDFPWPMLR--NEFKYFQRMTTTSSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPQGAHFLAKSLDDALKLIEQPELADKVDMVWIVGGSSVYKEAMNQPGHLRLFVTRIMQEFETDTFFPEIDLEKYKLLPEYPRVLPEVQEEKGIKYKFEVYEKKG---
DHFR_perMan   MVRPLNCIVAVSQNMGIGKNGDLPWPPLR--NEFKYFQRMTTTSSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPQGAHFLAKSLDDALKLIEQPELANKVDMVWIVGGSSVYKEAMNKPGHLRLFVTRIMQEFESDTFFPDIDLEKYKLLPEYPGVLSEVQEEEGIKYKFEVYEKKA---
DHFR_perPol   MVRPLNCIVAVSQNMGIGKNGDLPWPLLR--NEFKYFQRMTTTSSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPQGAHFLAKSLDDALKLIEQPELANKVDMVWVVGGSSVYKEAMNKPGHLRLFVTRIMQEFESDTFFPDIDLEKYKLLPEYPGVLSEVQEEEGIKYKFEVYEKKA---
DHFR_dipOrd   MGRSLNCIVAVSQNMGIGKNGDLPWPPLR--NEFKYFQRMTTTSTVEGKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPQGAHFLAKSLDEALKLIEQPELNNKVDMVWIVGGSSVYKEAMNQPGHHKLFVTRIMQEFESDTFFPEIDLEKYKLLPEYPGVLSEVQEEKGIKYKFEVYEKID---
DHFR_speTri   MVRPLNCIVAVSQNMGIGKNGDLPWPPLR--NEFKYFQRMMTTPSVEGKQNLVIIGRKTWFSIPEKNRPLNDRINIVLSRELKELPKGAHFLAKSLTDALKLTEQPELASKIDMIWIVGGSSVYKEAMIQPGHIKLFVTRIMQEFESDTFFPEIDLEKYKLLPEYPGILSEVQEEKGIKYKFEVYEKTD---
DHFR_cavPor   MVRPLNCIVAVSQNMGIGKNGDLPWPPLR--NEFGYFQRMTTTPSVEGKQNLVIMGRKTWFSLPEKSRPLKDRINVVLSRELKEPPQGAHFLAPSLDDALKLIEQPGLANEVDMVWIVGGSSVYKEAMERPGHIKLFVTRIMQEFESDTFFPEIDLKKYRLLPEYPGVPSEIQEEKGIKYKFEVYEKND---
DHFR_oryCun   MVRPLNCIVAVSQNMGIGKNGDLPWPPLR--NEFRYFQRMTTTSSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINVVLSRELKEPPQGAHFLAKSLADALKLIEEPELAEKVDMAWIVGGSSVHKEAMNQPGHLRLFVTRIIQEFESDTFFPEIDLEKYKLLPEYPGVLSDVQEEKGIKYKFEVYEKND---
DHFR_ochPri   MVRPLNCIVAVSQNMGIGRNGDLPWPPLR--NEFKYFQRMTTTSTVEGKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPPGAHFLAKSLDDALKLIEEPELAERVDMVWIVGGSSVYKEAMDKPGHLRLFVTRIMQEFESDTFFPEIDLEKYKLLPEYPGILPEVQEERGIKYRFEVYEKND---
DHFR_felCat   MVRPLNCIVAVSQNMGIGKNGDLPWPPLR--NEFKYFQRMTTTSSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRDLKEPPQGAHFLAKSLDDALELTEQPELAHKVDMVWVVGGSSVYKEAMNRPGHVRLFVTRIMQEFESDTFFPEIDLEKYKLLPEYPGVLSDIQEEKSIKYKFEVYEKNN---
DHFR_canFam   MVRTLNCIVAVSQNMGIGRNGDLPWPPLR--NEFKYFQRMTTNSSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRDLKEPPQGAHFLAKSLDDALKLTEQPELANKVDMVWIVGGSSVYKEAMNKPGHLRLFVTRIMHEFESDTFFPEIDLEKYKLLPEYPGVLSDVQEEKGIKYKFEVYEKND---
DHFR_vulVul   MVRTLNCIVAVSQNMGIGRNGTVPWPPLR--NEFKYFQRMTTNSSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRDLKEPPQGAHFLAKSLDDALKLTEQPELANKVDMVWIVGGSSVYKEAMNKPGHLRLFVTRIMHEFESDTFFPEIDLEKYKLLPEYPGVLSDVQEEKGIKYKFEVYEKND---
DHFR_musPut   MVRTLNCIAAVSQNMGIGKNGDLPWPPLR--NEFKFFQRMTTTSSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINLVLSRDLKEPPQGAHFLAKSLDDALKLIEQPELANKVDMVWIVGGSSVYKEAMNKPGHLRLFVTRIMQEFESDTFFPEIDLEKYKLLPEYPGVLSDVQEEKGIKYKFEVYEKND---
DHFR_ailMel   MVRTLNCIAAVSQNMGIGKNGDLPWPPLR--NEFKFFQRMTTTSSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINLVLSRDLKEPPQGAHFLAKSLDDALKLIEQPELANKVDIVWIVGGSSVYKEAMNKPGHLRLFVTRIMQEFESDTFFPEIDLDKYKLLPEYPGVPSDVQEEKGIKYKFEVYEKND---
DHFR_equCab   MVRPLNCIVAVSQNMGIGKNGDLPWPPLR--NEFKYFQRMTTASSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINLVLSRELKEPPQGAHFLAKSLDDALSLLEQPELANKVDMVWIVGGSSVYKEAMNKPGHLRLFVTRIMQEFESDTFFPEIDFEKYKLLPEYPGVLSDIQEEKGIKYKFEVYEKND---
DHFR_susScr   MVRPLNCIVAVSQNMGIGKNGDLPWPPLR--NEYKYFQRMTTTSSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPQGAHFLAKSLDDALKLTEQPELKNKVDMVWIVGGSSVYKEAMNKPGHIRLFVTRIMKEFESDTFFPEIDLEKYKLLSECSGVPSDVQEEKGIKYKFEVYEKNN---
DHFR_turTru   MVRPLNCIVAVSQNMGIGKNGDLPWPPLR--NEYRYFQRMTTTSSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPQGAHFLAKSLDGALKLIEQPELTNTVDMVWIVGGGSVYKEAMNKPGHLRLFVTRIMQEFESDTFFPEIDLEKYKLIPEYPGVPSDVQEEKGIKYKFEVYEKNN---
DHFR_oviAri   MVRPLNCIVAVSQNMGIGKNGNLPWPPLR--NEFKYFQRMTTVSSEEGKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPKGAHFLAKSLDDALELIEDPELTNKVDMVWIVGGSSVYKEAMNKPGHLRLFVTRIMQEFESDVFFPEIDFEKYKLLPEYPGVPLDVQEEKGIKYKFEVYEKNN---
DHFR_capHir   MVRPLNCIVAVSQNMGIGKNGNLPWPPLR--NEFKYFQRMTTVSSEEGKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPKGAHFLAKSLDDALELIEDPELTNKVDMVWIVGGSSVYKEAMNKPGHLRLFVTRIMQEFESDVFFPEIDFEKYKLLPEYPGVPSDVQEEKGIKYKFEVYEKNN---
DHFR_bosTau   MVRPLNCIVAVSQNMGIGKNGNLPWPPLR--NEFQYFQRMTTVSSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPKGAHFLAKSLDDALELIEDPELTNKVDVVWIVGGSSVYKEAMNKPGHVRLFVTRIMQEFESDAFFPEIDFEKYKLLPEYPGVPLDVQEEKGIKYKFEVYEKNN---
DHFR_myoLuc   MVRTLNCIAAVSQNMGIGKNGDLPWPPLR--NEFKYFQRMTTTSSVEGKQNLVIMGRKTWFSIPKKNRPLKDRINLVLSRELKEPPKGAHFLAKSLDDALKLIEEPELTNQVDMVWIVGGSSVYKEAMSKPGHQRLFVTRIMQEFESDTFFPEIDLEKYKLLPEYPGVLSDVQEEKGIKYKFEVYEKNE---
DHFR_pteVam   MVLSLNCIVAVSQNMGIGKNGDLPWPPLR--NEFKYFQRMTTTSSVEGKRNLVIMGRKTWFSIPKKNRPLKDRINLVLSRELKEPPKGAHFLAKSLDDALKLIEQPELRSKVDMVWIVGGSSVYKEAMNKPGHIKLFVTKIMQEFESDIFFPEIDLGKYKLLPEYPGVLSDVQEEKGIKYKFEVYEKND---
DHFR_sorAra   MVRPLNCIVAVSQNMGIGKNGELPWPPLR--NEFTYFRKMTTTSSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINLVLSRELKEPPTGAHFLAKSLDDALKLIEQPELTNKVDMVWIVGGSSVYQEVMNKPGHLRLFVTRIMQEFECDTFFPEIDLEKYKLLTDYPGVVSDVQEEKGIKYKFEVYEKKD---
DHFR_loxAfr   MVRPLNCIVAVSQNMGIGKNGDLP-PPLR--NEFKYFQRMTTTSSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINLVLSRGLKEPPQGAHFLAKSLDDALKLIEQPELRNKVDMVWIVGGSSVYKEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLEKYKRLSEYPGVLSDVQEEKGIKYKFEVYEKKD---
DHFR_proCap   MVRPLSCIVAVSQNMGIGKNGDLPWPPLR--NEFKHFRTMTSTPSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINLVLSRELKEPPQGAHFLAKSLDDALKLIEQPELTNKVDLVWIIGGSSVYKEAMSQPGHLTLFVTRIMQEFESDTFFPEIDLEKFKLLPEYPGVPSDVQEEKGIKYKFEVYEKKD---
DHFR_monDom   MVRQLNCIAAVSKNMGIGKDGDLPWPLLR--NEFKYFQKMTTTPSVEGKQNLVIMGKKTWFSIPEKCRPLKDRINVVLSRELKEPPQGAHYLASSLDNALNLLELPELADKVDMVWIVGGSSVYKEAMNKPGHQRLFITRILQEFESDTFFPEIDLKKYKQLQDYPGVSPDVQEEKGIQYKFEVYEKHE---
DHFR_macEug   MAPTINCIVAVAQNMGMGKNGDLPWPPLR--KEFQYFQKMTTTPSVEGKQNLLIMGKKTWFSIPEKNRPLKDRINLVLSRELKDPPLGAHYLAKSLDDALDLLRLPELANTVDMVWVVGGSSVYKEAMNKPGHQRLFVTRILQEFESDTFFPEIDLKRYKLLPEYPGVLTDVQEEKGIQHKCEIHEKHD---
DHFR_sarHar   MVRPINCIAAVSQNMGIGKNGDLPWPPLR--NEFNYFQKMTTTSSVEGKQNLLIMGKKTWFSIPEKHRPLKDRINLVLSRELKEPPPGAHYLAKSLDDALDLLELPELADKVDMIWVIGGSSVYKEAMEKPGHQRLFVTRILQEFECDTFFPEIDLKKYKLLPEYPGVLTDIQEEKGIQYKFEVYEKHD---
DHFR_triVul   MVRTINCIVAVSQNMGIGKNGDLPWPPLR--NDFKHFQKMTTIPSVEGKQNLLITGKKTWFSIPEKSRPLKDRINVVLSRELKEPPPGAHYLAKSLDDALDLLELPELANKVDMIWVVGGSSVYKEAMNKRGHNRLFVTRILQEFESDTFFPEIDLKRYKLLXEYPGVLTDVQEEKGIQYKFEVYEKHD---
DHFR_ornAna   MGPLLNCIVAVAKNGGIGNKGDLPWPALR--NEFRYFQKMTTTPTVKGKQNLVIMGKKTWFSIPEKSRPLKDRINIVLSRELKEPPQGAHYVAKSLDDALNLLHLPELASKVDMVWIIGGSSVYKEAMEKGGHQRLFVTRILQDFESDTFFPEIDLEKYKLIPEYPGVPADTQEEKGIQYRFEVYEKHI---
DHFR_tacAcu   MGRPLNCIAAVAKNGGIGNKGDLPWPPLR--NEYRYFQKMTTTPTVEGKENLVIMGKKTWFSIPEKSRPLKDRINIVLSRELKEPPQGAHYVAKSLDDALNLLNLPELASKVDMVWIIGGSSVYKEAMEKPGHQRLFVTRILQEFESDTFFPEIDLEKYKLIPEYPGVPADTQEEKGIQYRFEVYEKHI---
DHFR_galGal   MVRSLNSIVAVCQNMGIGKDGNLPWPPLR--NEYKYFQRMTSTSHVEGKQNAVIMGKKTWFSIPEKNRPLKDRINIVLSRELKEAPKGAHYLSKSLDDALALLDSPELKSKVDMVWIVGGTAVYKAAMEKPINHRLFVTRILHEFESDTFFPEIDYKDFKLLTEYPGVPADIQEENGIQYKFEVYQKSVLAQ
DHFR_lagLag   MVRSLNSIVAVCQNMGIGKDGNLPWPPLR--NEYKYFQRMTSTSHVEGKQNALIMGKKTWFSIPEKNRPLKDRINIVLSRELKEAPEGAHYLSKSLDDALALLDSPELKSKVDMVWIVGGTSVYKAAMEKPINHRLFVTRILHEFESDTFFPEIDYKDFKLLTEYPGVPADVQEENGIQYKFEVYQKSVLAQ
DHFR_anaPla   MVRSLNSIVAVCQNMGIGKDGNLPWPPLR--NEYKYFQRMTSTSHVEGKQNALIMGKKTWFSIPEKNRPLKDRINIVLSRELKEAPEGAHYLSKSLDDALALLDSPELKSKVDMVWIVGGTSVYKAAMEKPINHRLFVTRILHEFESDTFFPEIDYKDFKLLTEYPGVPADVQEENGIQYKFEVYEKTVLAQ
DHFR_taegut   MPRSLNSIVAISQNMGIGKDGRLPWPPLR--NDYKYFQRMTSASPVEGKKNAVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEAPKGAHYLSKSLDDALALLDSPELQSKVCMVWIVGGSSVYKEAMEKPIHHRLFVTRILKEFESDTFFPEIDHKDYKLLTEYPGVPADIQEENGIQYKFEVYEKTVVP-
DHFR_ficHyp   MVRSLNSIVAVSQNMGIGKDGRLPWPPLR--NEYKYFQRMTSTPRVEGKQNALIMGKKTWFSIPEKNRPLKDRINIVLSRELKEAPEGAHYLSKSLDDALALLDSPELKSKVDMVWIVGGTSVYKAAMEKPIHHRLFVTRILKEFESDTFFPEIDHKDYKLLTEYPGVPADIQEENGIQYKFEVYEKAVV--
DHFR_melUnd   MVRSLNSIVAVCQNMGIGKDGSLPWPPFR--NEYKYFQRMTSTSQVEGKQNALIMGKKTWFSIPEKNRPLKDRINIVLSRELQETPKGAHYLSKNLDDALALLDSPELKSKVDMVWIVGGTSVYKAAMEKPINHRLFVTRILQEFESDTFFPEIDHKDYKLLTEYPGVPADIQEENGIQYKFEVYEKAV---
DHFR_allMis   MVSSLNAIAAVSQNMGIGKNGTLPWPPLR--NEFKYFQRMTTTPSVPGKQNVLIMGKKTWFSIPEKNRPLKDRINIVLSRELAEVPKGAHHLARSLDDALVLLDSPELKNKIDLVWIVGGSSVYKAAMEKPINQRLFVTRILHEFESDTFFPEIDLEKYRLLPEFPGVPADVQEENGIHYKFEVYEKTV---
DHFR_croPor   MVSSLNAIAAVSQNMGIGKNGTLPWPPLR--NEFKYFQRMTTTPSVPGKQNVLIMGKKTWFSIPEKNRPLKDRINIVLSRELAEVPKGAHHLARSLDDALVLLDSPELKNKIDLVWIVGGSSVYKAAMEKPINQRLFVTRILHEFESDTFFPEIDLEKYRLLPEFPGVPADVQEENGIHYKFEVYEKTV---
DHFR_chrPic   MVRPLNCIAAVCQNMGIGKNGDLPWPPLR--NEFKYFQRMTTTTTVEGKQNVLIMGKRTWFSIPEKNRPLKDRINIVLSRELKETPKGAHYLAKSLDDALVLLDSPELKSKIDMVWIIGGSSIYKAAMEKPVHQRLFVTRILHEFESDTFFPEIDLEKYKHLPEYPGVPADVQEENGIRYKYEVYEKTV---
DHFR_anoCar   MVLSLNSIAAVCQNMGIGKNGQLPWPPLR--NEFKYFQKMTMTPTQEGKQNVVIMGKRTWFSIPEKNRPLKDRINIILSREMKEVPKGAHYLARSLDEALDLLQSPELENKIDLVWIVGGSSVYKAAMDKPIHHRLFVTRILHDFESDTFFPEIDLEKYKLIPSYAGVPTDIQEENGIRYRFEVYEKNI---
DHFR_pytMol   MVASLHSIVAVCNNMGIGKDGKLPWPPLR--NEFKHFQKMTMTTKEEGKQNVVIMGKKTWFSIPEKHRPLKNRINVVLSKELKDVPEGAHYLAKSLEEALDLLETSEMERKVDKVWIVGGSSVYKAAMEKPIHHQLFVTRIMHDFESDTFFPEIDLKKYKLLPKYTGIPTDIQEENGIQYKFEVYENTI---
DHFR_ambMex   MGRILNAIVAVCPSMGIGKDGNLPWPILR--NEFKYFQRMTMTATQEGKQNVVIMGRKTWLSIPEKNRPLKGRINIVLSRELTAPPPGAHYLANSLDGALELLETRELAGKVDLLWIIGGSSLYRELIGRPHHQRLFVTRILKEFECDTFLPEFDLEKYRLLPEYPGVSLGIQEENGIQYKFEVYEKVEEA-
DHFR_xenTro   MNPFLHAVVAVCPNQGIGKEGSLPWPLLR--NEFKHFQRLTMTPTVEDKKNVVIMGRKTWFSIPEKNRPLKERINIVLSKELKEPPTGAHYLSKSLAEAIDLLEDPELKDKVDLVWVIGGSSLYQELMEKPVNQRWFVTRILQEFECDTYLPEIDLNSFRLLPEYPGISPELQEENGVQYKFEVYEKIV---
DHFR_xenLae   MNQFLHAVVAVCPNQGIGKGGSLPWPLLR--NEFKHFQRLTMTPTVEGKKNVVIMGRKTWFSIPEKNRPLKERINIVLSKELKEPPTGAHYLSKSLTEAIDLLEDPELKDKVDLVWIIGGSSLYQELMEKPINQRWFVTRILQEFECDTYLPEINLNSFQLLPQYPGISPELQEENGVQYKFEVYEKIV---
DHFR_ranCat   MNQFLHAVVAVCPNQGIGKGGTLPWPLLR--NEFKHFQRLTMTPTTEGKQNVVIMGRRTWFSIPEKNRPLKGRINIVLSRELTEPPNGAHYLAKSLDEAVHLLKTPEMKEKIDLIWIIGGSSLYKEVMDKPVSQRLFITRIQQEFECDTFLPDVDLQSYKLLPEYPGISSEMQEENGIQYKFEVYEKML---
DHFR_latCha   MARLLNSIVAVCPNLGIGKDGNLPWHPKRLSNEFRYFQKMTTTPTVEGKQNVVIMGRKTWFSIPEKNRPLKGRVNVVLSRELQEAPEGAHYLARDLDSALKLLESPELADRVDLVWIIGGSALYKEALEHPSQHRLFVTRVLQQFECDTYLPEINFNKFTLLPEFPGVSSAIQEESGIQYKFEVYEKNS---
DHFR_gadMor   MSRVLNCIVAVCPDAGIGYKGDLPWHPTRLNNEFKHFRRLTVTPGAEDKQNVVIMGRKTWNSIPEKNRPLNDRINIVLSRQLEAPPSGAHHLARDLPSALRLLDVPE-GLLADQVWLIGGSSMYSEVMETPCPRRLFVTRILKQFPSDTFFPDISPAQFRLLPEYPGVSSELQEENGIQYRFQVYQSVDP--
DHFR_tetNig   MARVLNAIVAVCPDLGIGRNGDLPWHPIRLDNEFKHFRKMTSTPSVNGKQNVVIMGRKTWFSIPEKHRPLANRINIVLSRRSREPPAGAHHLAHDLSSALQLVET-QLAEQADQVWVIGGSSLYQEMMERPGTSRLFVTHVLKQFDCDTFLPEIRPQRYRLLPQFPGVPQELQEEKGIQYRYQVYESLEP--
DHFR_hipHip   MSRILNGIVAVCPDLGIGNRGNLPWHPVRLSNEFKHFRSMTATPSEKGKQNVVIMGRKTWFSIPEKNRPLNNRINIVLSRQCKAPPAGAHHLSADFSSALRLVDT-ELADQADQVWVIGGSSLYKEMMESPGTRRLFVTRILKQFDCDTFLPEISPDKYRQLPEFPGVPQELREENGIQYRFEVYESIEP--
DHFR_solSen   MSRVLNGIVAVCPDMGIGMTGNLPWHPVRLSNEFKHFRTMTATSSVKGKKNVVIMGRKTWFSIPEKNRPLNNRINIVLSRQCKLPPAGAHHLAADFSSALRLVET-ELADEADQIWIIGGSSLYKEMMESPGIKRLFVTRILKQFECDTFLPEISLDKYRLLPEFAGVSQELQEENAIQYRFEVYESVEQ--
DHFR_oreNil   MPRVLNAIVAVCPDRGIGNKGNLPWHPIRLSKEFAHFRKMTATPSVKGKQNVVIMGKKTWYSIPEKNRPLSNRINIVLSRECKVPPAGAHYLASDFSSALRLIDT-ELADQADQVWVIGGSSLYKEMMGSTGMRRLFVTQILKQFECDTFLPEISLDKYRLLPEFPDVPQELQEENGIQYRFKVYESIQE--
DHFR_dicLab   MSRILNGIVAVCPDLGIGMNGNLPWHPVRLGNEFKHFRTMTATPSVKGKQNVVIMGRKTWFSIPEKNRPLNNRINIVLSKECKVPPAGAHHLAADFNSALRLVET-ELADQADQVWVIGGSSLYKELMESPGTRRLFVTRILKQFECDTFLPEICPDKYQLLPEFPGVSQELQEENGIQYRFEVYESTEQ--
DHFR_perFla   MSRILNGIVAVCPDLGIGNNGNLPWHPVRLNNEFKHFRTMTATPSVNGKQNVVIMGRKTWFSIPEKNRPLNNRINIVLSRECKAPPAGAHHLARDFSSALRLVDT-ELAQQADTVWVIGGSSLYKEMMESPGTRRLFVTRILKQFECDTFLPEITPDKYRLLPEFPGVPQELQEENGIQYRFEVYESIQQ--
DHFR_spaAur   MSRIVNGIVAVCPDLGIGNNGNLPWHPVRLNREFKHFRTMTATPSVKGKQNVVIMGRKTWFSIPDKFRPLNNRINIVLSRECKVPPAGAHHLAADFSSALSLVDT-ELADQADQVWVIGGSSLYKELMESTRTSRLFVTRILKQFECDTFLPEISPDKYSLLPEFPGVPQGLQEENGIQYKFEVYERIKQ--
DHFR_gasAcu   MSRVLNGIVAVCPDLGIGCHGNLPWHPLRLSNEFKHFRRMTATASVKDKQNVVIMGRKTWFSIPEKNRPLNNRINIVLSRELKAPPAGAHHLAPDFSSALRLVDT-ELAEQADQVWVIGGSSLYKELLESPGTRRLFVTRILKQFECDTFLPEICPDRYRLLPEFPGVPEELQEENGVQYRFEVYESIDV--
DHFR_oryLat   MTRTLNGIVAVCPDLGIGKGGNLPWHPLRLSKDFALFRKMTSTPLVAGRQNVVIMGRKTWFSIPEKNRPLQNRINIVLSRQLKAPPAGAHFVAADFAAALRLVDS-ELAERADQVWIIGGSSIYXEMMESPAPRRLIVTRVQKQFECDTFFPEIRPEQFRLLPEFPGVSQELQEENGVSYKFEVYESIQQ--
DHFR_anoFim   MSRVLNAIVAVCPDLGIGRNGDLPWHPVRLNNEFKHFRRMTSTPSVEGKQNVVIMGRKTWFSIPEKNRPLNNRINIVLSRELKTPPAGAHHLAQDFNSALRLVDM-KLAEQVDQVWVIGGSSLYKELMESQGIRRLFVTQILKQFECDTFLPKISPDKYRLLPEFPGVPQELQEENGIQYRFEVYESIEQ--
DHFR_esoLuc   MSRVLNCIVAVCPNMGIGNKGNLPWHPKRLNNEFKYFQKMTMTPFVEGKQNVDIMGRKTWFSIPERNRPLKNRINIVLSRELKEPPPGAHHLASDFSSALRLLDTAEMAERVDQVWVIGGSSLYKEMMESEGIRRLFVTRILQQFESDTFLPEINQDKYRLLPEFPGVPQDVQEESGIQYRFEVYESVDH--
DHFR_salSal   MSRVPNCIVAVCPDVGIGNNGNLPWHPKRLNNEFKYFQKMTMTSSVEGKQNVVIMGRKTWFSIPERNRPLKNRINIVLSRELKKPPAGAHHLASDFRSALCLLDMAEMAERADQVWVIGGSSLYKEMMESQGTRRLFVTRILQQFKSDTFLPEINQDQYRLLPEFPGVPQDLQEEAGLQYRFEVYESIDQ--
DHFR_oncMyk   MSRVLNCIVAVCPDMGIGNNGNLPWHPKRLNNEFKYFQKMTMTSSVEGKQNAVIMGRKTWFSIPERNRPLKNRINIVLSRELKEPPAGAHQLASDFRSALRLLDTAEMAERVDQVWVIGGSSLYEEMMESQGTRRLFVTRILQQFKSDTFLPEINQDKYRLLPEFPGVPQDLQEEAGLQYRFEVYESIDQ--
DHFR_danRer   MSRILNCIVAVCPDMGIGKNGNLPWHPIRLSNELKHFQKMTMTPSDEGKKNVVIMGRKTWFSIPAAHRPLKNRINIVLSRELKTAPEGAHYLASDFSSALHLLDSGELEKLVDQVWIIGGSSLYKEVMERSGHRRLFVTRILKQFDCDTFIPNFDMDKYKLLPEFPGVPVGLQEDNGVQYLFEVYESIKH--
DHFR_cteIde   MSRILNCIVAVCPDMGIGRKGNLPWHPIRLSNEFKHFQKMTMTPSVEGKKNVVIMGRKTWFSIPAQNRPLKNRINIVLSRELKTAPEGAHYLASDLSSALHLLDSSELEKQVDQVWIIGGSALYKEVMERSGPRRLFVTRVLKQFDCDTFIPEINMDKYKLLPEFPGVPTGLQDENGVQYVFECMIVS----
DHFR_cypCar   MSRILNSIVAVCPDMGIGKNGNLPWHPIRLSNEFKHFQKMTMTPLVEGKKNVVLMGRKTWFSIPAANRPLKNRINIVLSRELKTAPEGAHYLASDFSSALHLLDSSELEKQVDQVWIIGGSALYKEVMESSGPRRLFVTRVLKQFDCDTFIPDINMDKYKLLPEFPGVPTGLQEENGVQYVFEVYESTDH--
DHFR_ictPun   MGRVLNCIVAVCPDMGIGRNGNLPWHPIRLSKEFKHFQKMTMTPTVEGKKNVVIMGRKTWFSIPAQNRPLKNRINVVLSRELKTAPEGAHYLAADFSSALRILENAELEAQADQVWIIGGSSLYKEVMESSAHQRLFVTRILKQFDCDTFIPNIDLDKFRLLPEFPGVPAGLQEENGLQYVFEVYESVEQ--
DHFR_leuEri   MTRLINSIVAVCPNMGIGNNGNFPWHPIRLSKEFKHFQRMTSTPSVEGKCNAVIMGRKTWHSIPEKNRPLKNRINIVLSRELKEVPEGAHYLAPDLDSALALLDSSDLQDKVDLVWIIGGSALYKEALESPASHRLFVTRILQDFDSDTFIPEINLNKFKLLPEFPDVPADIQEENGIRFKYEVYERISPV-
DHFR_squAca   MPRLVNCIVAVCPNMGIGKDGNFPWHPIRLSKELKHFQKMTATPSVEGKRNAVIMGRKTWFSIPEKHRPLKDRLNIVLSRELKELPHGANYLAHDLDSALAHLDSSELQDKVDLVWIIGGSTLYKEAMESPVSHRIFVTRILKDFESDTFLPEMDLNKFKLLPKFPDVPTEVQEDNGIQYKFEVYERIAHA-
DHFR_eptBur   MQNPVNVIAAVLPNMGIGWKGGLPWHSKSLVKEMKHFTRLTSAAA-EGKQNAVVMGRKTWESIPEKFRPLRNRINVVLSRTLSHLPLGAHYLATDFSNALAQLSESELCCRIDHIWIIGGSSIYKEALLHPACHRIFITRIMHDFEADTFFPHIDSKRFSLCQSYPGIPEEIQEENGVQYKFEVYEVVAK--
DHFR_petMar   -----------------------------------------------GKQNVVIMGRKTWFSIPEKKRPLKNRINIVLSRELKCLPEGAHYLVPDLNSALSLIDSPAIQEKLDGVWILGGGGVYKEAMEHSACRRLFITRVLQTMEADAFFPDIDADKFKLLP-----------------------------

                      10        20          30        40        50        60        70        80        90       100       110       120       130       140       150       160       170       180       190
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DHFR_homSap   MVGSLNCIVAVSQNMGIGKNGDLPWPPLR--NEFRYFQRMTTTSSVEGKQNLVIMGKKTWFSIPEKNRPLKGRINLVLSRELKEPPQGAHFLSRSLDDALKLTEQPELANKVDMVWIVGGSSVYKEAMNHPGHLKLFVTRIMQDFESDTFFPEIDLEKYKLLPEYPGVLSDVQEEKGIKYKFEVYEKND---
DHFR_panTro   .............................................................................................................S..................................................................................
DHFR_gorGor   .....................V..........................................................................................................................................................................
DHFR_ponAbe   .......................................................................D........................................................................................................................
DHFR_nomLeu   .................................................................................................................................Y..................................F...........................
DHFR_macMul   ..S.........................................................................................A.....................I...................................L.........................................
DHFR_papAnu   .AR............................S............................................................A.....................I.....N.............................L.........................................
DHFR_calJac   ..QL.................E.........................D........R..............D...V.............-L.A.....................I..I..........D..................................Q....P..D....................
DHFR_saiBol   ..QL....................................................R..............D...V...K............A.....................I..I..........D...........................K.....S.............................
DHFR_tarSyr   ..RP.Y.............D.T....................M.............R..............D...........V..P.....AK........I..........K......N...R...SQ................................S.............................
DHFR_otogar   ..RP..............................N......K..............R..............D...I........S.......AK........I.E........................Q...V............C.........K.....S.............................
DHF1_micMur   ..RT..................................K.................R.........S....D...V...............VAK........I..................A.......Q..........................K...................................
DHFR_tupBel   ..RP..............................K................I....R..............D....................AK........I.......Q.............R....Q..............................................................
DHFR_musMus   ..RP.........D...................WK.....................R..............D...I..........R.....AK......R.I......S..............Q....Q....R........E............G.............E................K....
DHFR_ratNor   ..RP......................L.......K.....................R..............D...I................AK........I......S......V.......Q....Q....R........E..........................EI...............K....
DHFR_criGri   ..RP..................F...M.......K.....................R..............D...I................AK........I......D...................Q....R........E..T...................R..PE................KG...
DHFR_perMan   ..RP..............................K.....................R..............D...I................AK........I..........................K....R........E........D.................E....E...........KA...
DHFR_perPol   ..RP......................L.......K.....................R..............D...I................AK........I.............V............K....R........E........D.................E....E...........KA...
DHFR_dipOrd   .GR...............................K.........T...........R..............D...I................AK...E....I.....N....................Q...H.........E..........................E................I....
DHFR_speTri   ..RP..............................K.....M..P..........I.R.............ND...I........L.K.....AK..T............S.I..I.............IQ...I.........E.......................I..E................T....
DHFR_cavPor   ..RP..............................G........P............R.....L...S....D...V................AP........I...G...E.................ER...I.........E............K..R........P.EI....................
DHFR_oryCun   ..RP....................................................R..............D...V................AK..A.....I.E....E....A........H.....Q....R......I.E................................................
DHFR_ochPri   ..RP..............R...............K.........T...........R..............D...I..........P.....AK........I.E....ER.................DK....R........E.......................I.PE....R....R...........
DHFR_felCat   ..RP..............................K.....................R..............D...I....D...........AK......E........H......V............R...VR........E...........................I....S...........N...
DHFR_canFam   ..RT..............R...............K.......N.............R..............D...I....D...........AK...................................K....R.......HE................................................
DHFR_vulVul   ..RT..............R..TV...........K.......N.............R..............D...I....D...........AK...................................K....R.......HE................................................
DHFR_musPut   ..RT....A.........................KF....................R..............D........D...........AK........I..........................K....R........E................................................
DHFR_ailMel   ..RT....A.........................KF....................R..............D........D...........AK........I..........I...............K....R........E............D...........P.......................
DHFR_equCab   ..RP..............................K.......A.............R..............D....................AK......S.L..........................K....R........E...........F...............I....................
DHFR_susScr   ..RP.............................YK.....................R..............D...I................AK..............K....................K...IR.......KE..................S.CS..P...................N...
DHFR_turTru   ..RP.............................Y......................R..............D...I................AK...G....I.....T.T.........G........K....R........E.................I......P...................N...
DHFR_oviAri   ..RP.................N............K.......V..E..........R..............D...I..........K.....AK......E.I.D...T....................K....R........E....V......F............PL..................N...
DHFR_capHir   ..RP.................N............K.......V..E..........R..............D...I..........K.....AK......E.I.D...T....................K....R........E....V......F............P...................N...
DHFR_bosTau   ..RP.................N............Q.......V.............R..............D...I..........K.....AK......E.I.D...T....V...............K...VR........E....A......F............PL..................N...
DHFR_myoLuc   ..RT....A.........................K.....................R.......K......D..............K.....AK........I.E...T.Q.................SK...QR........E............................................E...
DHFR_pteVam   ..L...............................K..............R......R.......K......D..............K.....AK........I.....RS...................K...I.....K...E....I.......G...................................
DHFR_sorAra   ..RP.................E............T..RK.................R..............D..............T.....AK........I.....T...............Q.V..K....R........E..C...............TD....V..................K....
DHFR_loxAfr   ..RP....................-.........K.....................R..............D........G...........AK........I.....R....................Q....R........E................R.S........................K....
DHFR_proCap   ..RP.S............................KH.RT..S.P............R..............D....................AK........I.....T....L...I..........SQ....T........E..............F.........P..................K....
DHFR_monDom   ..RQ....A...K......D......L.......K...K....P......................C....D...V..............Y.AS...N..N.L.L....D...................K...QR..I...L.E............K...Q.QD....SP........Q........HE...
DHFR_macEug   .APTI......A....M..............K..Q...K....P........L..................D...........D..L...Y.AK......D.LRL.....T.....V............K...QR......L.E............KR...........T........QH.C.IH..H....
DHFR_sarHar   ..RPI...A.........................N...K.............L.............H....D..............P...Y.AK......D.L.L....D....I.VI..........EK...QR......L.E..C.........K............T.I......Q........H....
DHFR_triVul   ..RTI...........................D.KH..K...IP........L.T...........S....D...V..........P...Y.AK......D.L.L.........I.V............KR..NR......L.E............KR....X......T........Q........H....
DHFR_ornAna   .GPL.......AK.G...NK......A...........K....PT.K...................S....D...I..............YVAK......N.LHL....S.......I..........EKG..QR......L...................I......PA.T......Q.R......HI...
DHFR_tacAcu   .GRP....A..AK.G...NK.............Y....K....PT....E................S....D...I..............YVAK......N.LNL....S.......I..........EK...QR......L.E.................I......PA.T......Q.R......HI...
DHFR_galGal   ..R...S....C.......D.N...........YK......S..H......A...................D...I........A.K...Y..K......A.LDS...KS..........TA...A..EK.INHR......LHE...........YKDF...T.....PA.I...N..Q......Q.SVLAQ
DHFR_lagLag   ..R...S....C.......D.N...........YK......S..H......AL..................D...I........A.E...Y..K......A.LDS...KS..........T....A..EK.INHR......LHE...........YKDF...T.....PA.....N..Q......Q.SVLAQ
DHFR_anaPla   ..R...S....C.......D.N...........YK......S..H......AL..................D...I........A.E...Y..K......A.LDS...KS..........T....A..EK.INHR......LHE...........YKDF...T.....PA.....N..Q........TVLAQ
DHFR_taegut   .PR...S...I........D.R..........DYK......SA.P....K.A....R..............D...I........A.K...Y..K......A.LDS...QS..C...............EK.I.HR......LKE...........HKD....T.....PA.I...N..Q........TVVP.
DHFR_ficHyp   ..R...S............D.R...........YK......S.PR......AL..................D...I........A.E...Y..K......A.LDS...KS..........T....A..EK.I.HR......LKE...........HKD....T.....PA.I...N..Q........AVV..
DHFR_melUnd   ..R...S....C.......D.S.....F.....YK......S..Q......AL..................D...I......Q.T.K...Y..KN.....A.LDS...KS..........T....A..EK.INHR......L.E...........HKD....T.....PA.I...N..Q........AV...
DHFR_allMis   ..S...A.A............T............K........P..P....VL..................D...I......A.V.K...H.A.......V.LDS...K..I.L...........A..EK.INQR......LHE...............R....F...PA.....N..H........TV...
DHFR_croPor   ..S...A.A............T............K........P..P....VL..................D...I......A.V.K...H.A.......V.LDS...K..I.L...........A..EK.INQR......LHE...............R....F...PA.....N..H........TV...
DHFR_chrPic   ..RP....A..C......................K........TT......VL....R.............D...I........T.K...Y.AK......V.LDS...KS.I.....I....I..A..EK.V.QR......LHE................H.......PA.....N..R..Y.....TV...
DHFR_anoCar   ..L...S.A..C.........Q............K...K..M.PTQ.....V.....R.............D...II....M..V.K...Y.A....E..D.LQS...E..I.L...........A..DK.I.HR......LH..................I.S.A..PT.I...N..R.R.......I...
DHFR_pytMol   ..A..HS....CN......D.K............KH..K..M.TKE.....V..............H....N...V...K...DV.E...Y.AK..EE..D.L.TS.MER...K...........A..EK.I.HQ.......H.............K......K.T.IPT.I...N..Q.......NTI...
DHFR_ambMex   .GRI..A....CPS.....D.N....I.......K......M.ATQ.....V....R...L..............I......TA..P...Y.AN...G..E.L.TR...G...LL..I....L.R.LIGR.H.QR......LKE..C...L..F.....R........SLGI...N..Q........VEEA.
DHFR_xenTro   .NPF.HAV...CP.Q....E.S....L.......KH...L.M.PT..D.K.V....R..............E...I...K......T...Y..K..AE.ID.L.D...KD...L..VI....L.Q.L.EK.VNQRW.....L.E..C..YL.....NSFR.......ISPEL...N.VQ........IV...
DHFR_xenLae   .NQF.HAV...CP.Q....G.S....L.......KH...L.M.PT....K.V....R..............E...I...K......T...Y..K..TE.ID.L.D...KD...L...I....L.Q.L.EK.INQRW.....L.E..C..YL...N.NSFQ...Q...ISPEL...N.VQ........IV...
DHFR_ranCat   .NQF.HAV...CP.Q....G.T....L.......KH...L.M.PTT.....V....RR.................I......T...N...Y.AK...E.VH.LKT..MKE.I.LI..I....L...V.DK.VSQR..I...Q.E..C...L.DV..QS.........IS.EM...N..Q........ML...
DHFR_latCha   .ARL..S....CP.L....D.N...H.K.LS.......K....PT......V....R................V.V......Q.A.E...Y.A.D..S....L.S....DR..L...I...AL....LE..SQHR.....VL.Q..C..YL...NFN.FT....F...S.AI...S..Q.........S...
DHFR_gadMor   .SRV.......CPDA...YK.....H.T.LN...KH.R.L.V.PGA.D...V....R...N.........ND...I....Q.EA..S...H.A.D.PS..R.LDV..-GLLA.Q..LI....M.S.V.ET.CPRR......LKQ.P......D.SPAQFR........S.EL...N..Q.R.Q..QSV.P..
DHFR_tetNig   .ARV..A....CPDL...R......H.I.LD...KH.RK..S.P..N....V....R.........H...AN...I....RSR...A...H.AHD.SS..Q.V.T-Q..EQA.Q..VI....L.Q.M.ER..TSR....HVLKQ.DC...L...RPQR.R...QF...PQEL......Q.RYQ...SLEP..
DHFR_hipHip   .SRI..G....CPDL...NR.N...H.V.LS...KH.RS..A.P.EK....V....R.............NN...I....QC.A..A...H..ADFSS..R.VDT-...DQA.Q..VI....L...M.ES..TRR......LKQ.DC...L...SPD..RQ...F...PQELR..N..Q.R.....SIEP..
DHFR_solSen   .SRV..G....CPD....MT.N...H.V.LS...KH.RT..A....K..K.V....R.............NN...I....QC.L..A...H.AADFSS..R.V.T-...DEA.QI..I....L...M.ES..IKR......LKQ..C...L...S.D..R....FA..SQEL...NA.Q.R.....SVEQ..
DHFR_oreNil   .PRV..A....CPDR...NK.N...H.I.LSK..AH.RK..A.P..K....V........Y.........SN...I.....C.V..A...Y.ASDFSS..R.IDT-...DQA.Q..VI....L...M.GST.MRR....Q.LKQ..C...L...S.D..R....F.D.PQEL...N..Q.R.K...SIQE..
DHFR_dicLab   .SRI..G....CPDL...M..N...H.V.LG...KH.RT..A.P..K....V....R.............NN...I...K.C.V..A...H.AADFNS..R.V.T-...DQA.Q..VI....L...L.ES..TRR......LKQ..C...L...CPD..Q....F...SQEL...N..Q.R.....STEQ..
DHFR_perFla   .SRI..G....CPDL...N..N...H.V.LN...KH.RT..A.P..N....V....R.............NN...I.....C.A..A...H.A.DFSS..R.VDT-...QQA.T..VI....L...M.ES..TRR......LKQ..C...L...TPD..R....F...PQEL...N..Q.R.....SIQQ..
DHFR_spaAur   .SRIV.G....CPDL...N..N...H.V.LNR..KH.RT..A.P..K....V....R.......D.F...NN...I.....C.V..A...H.AADFSS..S.VDT-...DQA.Q..VI....L...L.ESTRTSR......LKQ..C...L...SPD..S....F...PQGL...N..Q.......RIKQ..
DHFR_gasAcu   .SRV..G....CPDL...CH.N...H...LS...KH.R...A.A..KD...V....R.............NN...I.......A..A...H.APDFSS..R.VDT-...EQA.Q..VI....L...LLES..TRR......LKQ..C...L...CPDR.R....F...PEEL...N.VQ.R.....SI.V..
DHFR_oryLat   .TRT..G....CPDL....G.N...H...LSKD.AL.RK..S.PL.A.R..V....R.............QN...I....Q..A..A....VAADFAA..R.VDS-...ERA.Q...I....I.X.M.ES.APRR.I...VQKQ..C.......RP.QFR....F...SQEL...N.VS.......SIQQ..
DHFR_anoFim   .SRV..A....CPDL...R......H.V.LN...KH.R...S.P.......V....R.............NN...I.......T..A...H.AQDFNS..R.VDM-K..EQ..Q..VI....L...L.ESQ.IRR....Q.LKQ..C...L.K.SPD..R....F...PQEL...N..Q.R.....SIEQ..
DHFR_esoLuc   .SRV.......CP.....NK.N...H.K.LN...K...K..M.PF......VD...R........R.....N...I..........P...H.ASDFSS..R.LDTA.M.ER..Q..VI....L...M.ESE.IRR......L.Q......L...NQD..R....F...PQ.....S..Q.R.....SV.H..
DHFR_salSal   .SRVP......CPDV...N..N...H.K.LN...K...K..M.........V....R........R.....N...I.......K..A...H.ASDFRS..C.LDMA.M.ERA.Q..VI....L...M.ESQ.TRR......L.Q.K....L...NQDQ.R....F...PQ.L...A.LQ.R.....SI.Q..
DHFR_oncMyk   .SRV.......CPD....N..N...H.K.LN...K...K..M.........A....R........R.....N...I..........A...Q.ASDFRS..R.LDTA.M.ER..Q..VI....L.E.M.ESQ.TRR......L.Q.K....L...NQD..R....F...PQ.L...A.LQ.R.....SI.Q..
DHFR_danRer   .SRI.......CPD.......N...H.I.LS..LKH..K..M.P.D...K.V....R.......AAH....N...I.......TA.E...Y.ASDFSS..H.LDSG..EKL..Q...I....L...V.ERS..RR......LKQ.DC...I.NF.MD.......F...PVGL..DN.VQ.L.....SIKH..
DHFR_cteIde   .SRI.......CPD....RK.N...H.I.LS...KH..K..M.P.....K.V....R.......AQ.....N...I.......TA.E...Y.ASD.SS..H.LDSS..EKQ..Q...I...AL...V.ERS.PRR.....VLKQ.DC...I...NMD.......F...PTGL.D.N.VQ.V..CMIVS-...
DHFR_cypCar   .SRI..S....CPD.......N...H.I.LS...KH..K..M.PL....K.V.L..R.......AA.....N...I.......TA.E...Y.ASDFSS..H.LDSS..EKQ..Q...I...AL...V.ESS.PRR.....VLKQ.DC...I.D.NMD.......F...PTGL...N.VQ.V.....ST.H..
DHFR_ictPun   .GRV.......CPD....R..N...H.I.LSK..KH..K..M.PT....K.V....R.......AQ.....N...V.......TA.E...Y.AADFSS..RIL.NA..EAQA.Q...I....L...V.ESSA.QR......LKQ.DC...I.N...D.FR....F...PAGL...N.LQ.V.....SVEQ..
DHFR_leuEri   .TRLI.S....CP.....N..NF..H.I.LSK..KH.....S.P.....C.A....R...H..........N...I........V.E...Y.APD..S..A.LDSSD.QD...L...I...AL....LES.ASHR......L...D....I...N.N.F.....F.D.PA.I...N..RF.Y....RISPV.
DHFR_squAca   .PRLV......CP......D.NF..H.I.LSK.LKH..K..A.P.....R.A....R.........H....D.L.I........L.H..NY.AHD..S..AHLDSS..QD...L...I...TL.....ES.VSHRI.....LK.......L..M..N.F....KF.D.PTE...DN..Q.......RIAHA.
DHFR_eptBur   .QNPV.V.A..LP.....WK.G...HSKSLVK.MKH.T.L.SAAA-.....A.V..R...E.....F...RN...V....T.SHL.L...Y.ATDFSN..AQLSES..CCRI.HI..I....I....LL..ACHRI.I....H...A.....H..SKRFS.CQS...IPEEI...N.VQ.......VVAK..
DHFR_petMar   -----------------------------..----------------....V....R.........K....N...I.......CL.E...Y.VPD.NS..S.IDS.AIQE.L.G...L..GG......E.SACRR..I..VL.TM.A.A...D..AD.F....--------------------------...

homSap  PWPPLR--NE  TTSSVEGKQNL  SIPEKNRP  LTEQPELANKV  AMNHPG  KLLPEYPGVLSD
xenTro  PWPLLR--NE  MTPTVEDKKNV  SIPEKNRP  LLEDPELKDKV  LMEKPV  RLLPEYPGISPE Identity: 36/55 (65%) Similar: 42/55 (76%)
latCha  PWHPKRLSNE  TTPTVEGKQNV  SIPEKNRP  LLESPELADRV  ALEHPS  TLLPEFPGVSSA Identity: 40/57 (70%) Similar: 46/57 (80%)
danRer  PWHPIRLSNE  MTPSDEGKKNV  SIPAAHRP  LLDSGELEKLV  VMERSG  KLLPEFPGVPVG Identity: 31/55 (56%) Similar: 37/55 (67%)
esoLuc  PWHPKRLNNE  MTPFVEGKQNV  SIPERNRP  LLDTAEMAERV  MMESEG  RLLPEFPGVPQD
salSal  PWHPKRLNNE  MTSSVEGKQNV  SIPERNRP  LLDMAEMAERA  MMESQG  RLLPEFPGVPQD
oncMyk  PWHPKRLNNE  MTSSVEGKQNA  SIPERNRP  LLDTAEMAERV  MMESQG  RLLPEFPGVPQD
cteIde  PWHPIRLSNE  MTPSVEGKKNV  SIPAQNRP  LLDSSELEKQV  VMERSG  KLLPEFPGVPTG
cypCar  PWHPIRLSNE  MTPLVEGKKNV  SIPAANRP  LLDSSELEKQV  VMESSG  KLLPEFPGVPTG
ictPun  PWHPIRLSKE  MTPTVEGKKNV  SIPAQNRP  ILENAELEAQA  VMESSA  RLLPEFPGVPAG
gadMor  PWHPTRLNNE  VTPGAEDKQNV  SIPEKNRP  LLDVPE-GLLA  VMETPC  RLLPEYPGVSSE
tetNig  PWHPIRLDNE  STPSVNGKQNV  SIPEKHRP  LVET-QLAEQA  MMERPG  RLLPQFPGVPQE
hipHip  PWHPVRLSNE  ATPSEKGKQNV  SIPEKNRP  LVDT-ELADQA  MMESPG  RQLPEFPGVPQE
solSen  PWHPVRLSNE  ATSSVKGKKNV  SIPEKNRP  LVET-ELADEA  MMESPG  RLLPEFAGVSQE
oreNil  PWHPIRLSKE  ATPSVKGKQNV  SIPEKNRP  LIDT-ELADQA  MMGSTG  RLLPEFPDVPQE
dicLab  PWHPVRLGNE  ATPSVKGKQNV  SIPEKNRP  LVET-ELADQA  LMESPG  QLLPEFPGVSQE
perFla  PWHPVRLNNE  ATPSVNGKQNV  SIPEKNRP  LVDT-ELAQQA  MMESPG  RLLPEFPGVPQE
spaAur  PWHPVRLNRE  ATPSVKGKQNV  SIPDKFRP  LVDT-ELADQA  LMESTR  SLLPEFPGVPQG
gasAcu  PWHPLRLSNE  ATASVKDKQNV  SIPEKNRP  LVDT-ELAEQA  LLESPG  RLLPEFPGVPEE
oryLat  PWHPLRLSKD  STPLVAGRQNV  SIPEKNRP  LVDS-ELAERA  MMESPA  RLLPEFPGVSQE
anoFim  PWHPVRLNNE  STPSVEGKQNV  SIPEKNRP  LVDM-KLAEQV  LMESQG  RLLPEFPGVPQE
leuEri  PWHPIRLSKE  STPSVEGKCNA  SIPEKNRP  LLDSSDLQDKV  ALESPA  KLLPEFPDVPAD
squAca  PWHPIRLSKE  ATPSVEGKRNA  SIPEKHRP  HLDSSELQDKV  AMESPV  KLLPKFPDVPTE

homSap  PWPPLR--NE  TTSSVEGKQNL  SIPEKNRP  LTEQPELANKV  AMNHPG  KLLPEYPGVLSD
xenTro  ...L..--..  M.PT..D.K.V  ........  .L.D...KD..  L.EK.V  R.......ISPE
latCha  ..H.K.LS..  ..PT......V  ........  .L.S....DR.  .LE..S  T....F...S.A
danRer  ..H.I.LS..  M.P.D...K.V  ...AAH..  .LDSG..EKL.  V.ERS.  .....F...PVG
esoLuc  ..H.K.LN..  M.PF......V  ....R...  .LDTA.M.ER.  M.ESE.  R....F...PQ.
salSal  ..H.K.LN..  M.........V  ....R...  .LDMA.M.ERA  M.ESQ.  R....F...PQ.
oncMyk  ..H.K.LN..  M.........A  ....R...  .LDTA.M.ER.  M.ESQ.  R....F...PQ.
cteIde  ..H.I.LS..  M.P.....K.V  ...AQ...  .LDSS..EKQ.  V.ERS.  .....F...PTG
cypCar  ..H.I.LS..  M.PL....K.V  ...AA...  .LDSS..EKQ.  V.ESS.  .....F...PTG
ictPun  ..H.I.LSK.  M.PT....K.V  ...AQ...  IL.NA..EAQA  V.ESSA  R....F...PAG
gadMor  ..H.T.LN..  V.PGA.D...V  ........  .LDV..-GLLA  V.ET.C  R........S.E
tetNig  ..H.I.LD..  S.P..N....V  .....H..  .V.T-Q..EQA  M.ER..  R...QF...PQE
hipHip  ..H.V.LS..  A.P.EK....V  ........  .VDT-...DQA  M.ES..  RQ...F...PQE
solSen  ..H.V.LS..  A....K..K.V  ........  .V.T-...DEA  M.ES..  R....FA..SQE
oreNil  ..H.I.LSK.  A.P..K....V  ........  .IDT-...DQA  M.GST.  R....F.D.PQE
dicLab  ..H.V.LG..  A.P..K....V  ........  .V.T-...DQA  L.ES..  Q....F...SQE
perFla  ..H.V.LN..  A.P..N....V  ........  .VDT-...QQA  M.ES..  R....F...PQE
spaAur  ..H.V.LNR.  A.P..K....V  ...D.F..  .VDT-...DQA  L.ESTR  S....F...PQG
gasAcu  ..H...LS..  A.A..KD...V  ........  .VDT-...EQA  LLES..  R....F...PEE
oryLat  ..H...LSKD  S.PL.A.R..V  ........  .VDS-...ERA  M.ES.A  R....F...SQE
anoFim  ..H.V.LN..  S.P.......V  ........  .VDM-K..EQ.  L.ESQ.  R....F...PQE
leuEri  ..H.I.LSK.  S.P.....C.A  ........  .LDSSD.QD..  .LES.A  .....F.D.PA.
squAca  ..H.I.LSK.  A.P.....R.A  .....H..  HLDSS..QD..  ..ES.V  ....KF.D.PTE

Vertebrate reference sequences

>DHFR_homSap Homo sapiens (human) AC130896 tightly syntentic -+ MSH3 chr5
0 MVGSLNCIVAVSQNMGIGKNGDLPWPPLR 2
1 NEFRYFQRMTTTSSVE 1
2 GKQNLVIMGKKTWFSIPEKNRPLKGRINLVLSRELK 2
1 EPPQGAHFLSRSLDDALKLTEQPELANKVDMVWIVGGSSVYK 0
0 EAMNHPGHLKLFVTRIMQDFESDTFFPEIDLEKYKLLPE 2
1 YPGVLSDVQEEKGIKYKFEVYEKND* 0

>DHFR_panTro Pan troglodytes (chimp) XM_001138522
MVGSLNCIVAVSQNMGIGKNGDLPWPPLRNEFRYFQRMTTTSSVEGKQNLVIMGKKTWFSIPEKNRPLKGRINLVLSRELKEPPQGAHFLSRSLDDALKLTEQPELASKVDMVWIVGGSSVYKEAMNHPGHLKLFVTRIMQDFESDTFFPEIDLEKYKLLPEYPGVLSDVQEEKGIKYKFEVYEKND

>DHFR_gorGor Gorilla gorilla (gorilla) CU104654
MVGSLNCIVAVSQNMGIGKNGVLPWPPLRNEFRYFQRMTTTSSVEGKQNLVIMGKKTWFSIPEKNRPLKGRINLVLSRELKEPPQGAHFLSRSLDDALKLTEQPELANKVDMVWIVGGSSVYKEAMNHPGHLKLFVTRIMQDFESDTFFPEIDLEKYKLLPEYPGVLSDVQEEKGIKYKFEVYEKND

>DHFR_ponAbe Pongo abelii (orangutan) XM_002815704 syntenic 
MVGSLNCIVAVSQNMGIGKNGDLPWPPLRNEFRYFQRMTTTSSVEGKQNLVIMGKKTWFSIPEKNRPLKDRINLVLSRELKEPPQGAHFLSRSLDDALKLTEQPELANKVDMVWIVGGSSVYKEAMNHPGHLKLFVTRIMQDFESDTFFPEIDLEKYKLLPEYPGVLSDVQEEKGIKYKFEVYEKND

>DHFR_nomLeu Nomascus leucogenys (gibbon) XM_003261693
MVGSLNCIVAVSQNMGIGKNGDLPWPPLRNEFRYFQRMTTTSSVEGKQNLVIMGKKTWFSIPEKNRPLKGRINLVLSRELKEPPQGAHFLSRSLDDALKLTEQPELANKVDMVWIVGGSSVYKEAMNYPGHLKLFVTRIMQDFESDTFFPEIDLEKYKLLPEFPGVLSDVQEEKGIKYKFEVYEKND

>DHFR_macMul Macaca mulatta (rhesus) XM_001110551 syntenic
MVSSLNCIVAVSQNMGIGKNGDLPWPPLRNEFRYFQRMTTTSSVEGKQNLVIMGKKTWFSIPEKNRPLKGRINLVLSRELKEPPQGAHFLARSLDDALKLTEQPELANKVDMIWIVGGSSVYKEAMNHPGHLKLFVTRIMQDFESDTFLPEIDLEKYKLLPEYPGVLSDVQEEKGIKYKFEVYEKND

>DHFR_papAnu Papio anubis (baboon) FC175564
MARSLNCIVAVSQNMGIGKNGDLPWPPLRSEFRYFQRMTTTSSVEGKQNLVIMGKKTWFSIPEKNRPLKGRINLVLSRELKEPPQGAHFLARSLDDALKLTEQPELANKVDMIWIVGGNSVYKEAMNHPGHLKLFVTRIMQDFESDTFLPEIDLEKYKLLPEYPGVLSDVQEEKGIKYKFEVYEKND

>DHFR_calJac Callithrix jacchus (marmoset) XM_002745986 tightly syntentic -+ MSH3 4 recent pseudogenes
MVQLLNCIVAVSQNMGIGKNGELPWPPLRNEFRYFQRMTTTSSVEDKQNLVIMGRKTWFSIPEKNRPLKDRINVVLSRELKEPPQGALLARSLDDALKLTEQPELANKVDMIWIIGGSSVYKEAMDHPGHLKLFVTRIMQDFESDTFFPEIDLEKYKLLPQYPGVPSDDQEEKGIKYKFEVYEKND 

>DHFR_saiBol Saimiri boliviensis (squirrel_monkey) AGCE01006205/AGCE01053749 numerous pseudogenes syntentic MSH3
MVQLLNCIVAVSQNMGIGKNGDLPWPPLRNEFRYFQRMTTTSSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINVVLSKELKEPPQGAHFLARSLDDALKLTEQPELANKVDMIWIIGGSSVYKEAMDHPGHLKLFVTRIMQDFESDTFFPEIDlkkykllseYPGVLSDVQEEKGIKYKFEVYEKND

>DHFR_tarSyr Tarsius syrichta (tarsier) ABRT010238682/ABRT010238686/ABRT010468683
MVRPLYCIVAVSQNMGIGKDGTLPWPPLRNEFRYFQRMTTMSSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINLVLSRELKVPPPGAHFLAKSLDDALKLIEQPELANKVDKVWIVGGNSVYREAMSQPGHLKLFVTRIMQDFESDTFFPEIDLEKYKLLSEYPGVLSDVQEEKGIKYKFEVYEKND

>DHFR_otogar Otolemur garnettii (bushbaby) AAQR03040271/AAQR03040268
MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFNYFQRMTKTSSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKESPQGAHFLAKSLDDALKLIEEPELANKVDMVWIVGGSSVYKEAMNQPGHVKLFVTRIMQDFECDTFFPEIDLKKYKLLSEYPGVLSDVQEEKGIKYKFEVYEKND

>DHF1_micMur Microcebus murinus (lemur) wgs Blat
MVRTLNCIVAVSQNMGIGKNGDLPWPPLRNEFRYFQKMTTTSSVEGKQNLVIMGRKTWFSIPEKSRPLKDRINVVLSRELKEPPQGAHFVAKSLDDALKLIEQPELANKVDMVWIVGGSAVYKEAMNQPGHLKLFVTRIMQDFESDTFFPEIDLKKYKLLPE YPGVLSDVQEEKGIKYKFEVYEKND

>DHFR_tupBel Tupaia belangeri (treeshrew) AAPY01231248/AAPY01231253/AAPY01729648/SRA|SRR005484.246484.2
MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVEGKQNIVIMGRKTWFSIPEKNRPLKDRINLVLSRELKEPPQGAHFLAKSLDDALKLIEQPELANQVDMVWIVGGSSVYREAMNQPGHLKLFVTRIMQDFESDTFFPEIDLEKYKLLPE

>DHFR_musMus Mus musculus (mouse) AK031241
MVRPLNCIVAVSQDMGIGKNGDLPWPPLRNEWKYFQRMTTTSSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQPELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEYPGVLSEVQEEKGIKYKFEVYEKKD

>DHFR_ratNor Rattus norvegicus (rat) NM_130400
MVRPLNCIVAVSQNMGIGKNGDLPWPLLRNEFKYFQRMTTTSSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPQGAHFLAKSLDDALKLIEQPELASKVDMVWVVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLEKYKLLPEYPGVLSEIQEEKGIKYKFEVYEKKD

>DHFR_criGri Cricetulus griseus (hamster) NM_001244016
MVRPLNCIVAVSQNMGIGKNGDFPWPMLRNEFKYFQRMTTTSSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPQGAHFLAKSLDDALKLIEQPELADKVDMVWIVGGSSVYKEAMNQPGHLRLFVTRIMQEFETDTFFPEIDLEKYKLLPEYPRVLPEVQEEKGIKYKFEVYEKKG

>DHFR_perMan Peromyscus maniculatus (deermouse) GH506523
MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPQGAHFLAKSLDDALKLIEQPELANKVDMVWIVGGSSVYKEAMNKPGHLRLFVTRIMQEFESDTFFPDIDLEKYKLLPEYPGVLSEVQEEEGIKYKFEVYEKKA

>DHFR_perPol Peromyscus polionotus (deermouse) GH462222
MVRPLNCIVAVSQNMGIGKNGDLPWPLLRNEFKYFQRMTTTSSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPQGAHFLAKSLDDALKLIEQPELANKVDMVWVVGGSSVYKEAMNKPGHLRLFVTRIMQEFESDTFFPDIDLEKYKLLPEYPGVLSEVQEEEGIKYKFEVYEKKA

>DHFR_dipOrd Dipodomys ordii (kangaroo_rat) wgsBlast
MGRSLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSTVEGKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPQGAHFLAKSLDEALKLIEQPELNNKVDMVWIVGGSSVYKEAMNQPGHHKLFVTRIMQEFESDTFFPEIDLEKYKLLPEYPGVLSEVQEEKGIKYKFEVYEKID

>DHFR_speTri Spermophilus tridecemlineatus (ground_squirrel) ti|1005446275
MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMMTTPSVEGKQNLVIIGRKTWFSIPEKNRPLNDRINIVLSRELKELPKGAHFLAKSLTDALKLTEQPELASKIDMIWIVGGSSVYKEAMIQPGHIKLFVTRIMQEFESDTFFPEIDLEKYKLLPEYPGILSEVQEEKGIKYKFEVYEKTD

>DHFR_cavPor Cavia porcellus (guinea_pig) XM_003479518 Hystricognathi
MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFGYFQRMTTTPSVEGKQNLVIMGRKTWFSLPEKSRPLKDRINVVLSRELKEPPQGAHFLAPSLDDALKLIEQPGLANEVDMVWIVGGSSVYKEAMERPGHIKLFVTRIMQEFESDTFFPEIDLKKYRLLPEYPGVPSEIQEEKGIKYKFEVYEKND

>DHFR_oryCun Oryctolagus cuniculus (rabbit) FJ687582
MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFRYFQRMTTTSSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINVVLSRELKEPPQGAHFLAKSLADALKLIEEPELAEKVDMAWIVGGSSVHKEAMNQPGHLRLFVTRIIQEFESDTFFPEIDLEKYKLLPEYPGVLSDVQEEKGIKYKFEVYEKND

>DHFR_ochPri Ochotona princeps (pika) wgsBlast/ti|1497151195
MVRPLNCIVAVSQNMGIGRNGDLPWPPLRNEFKYFQRMTTTSTVEGKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPPGAHFLAKSLDDALKLIEEPELAERVDMVWIVGGSSVYKEAMDKPGHLRLFVTRIMQEFESDTFFPEIDLEKYKLLPEYPGILPEVQEERGIKYRFEVYEKND

>DHFR_felCat Felis catus (cat) wgsBlast
MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRDLKEPPQGAHFLAKSLDDALELTEQPELAHKVDMVWVVGGSSVYKEAMNRPGHVRLFVTRIMQEFESDTFFPEIDLEKYKLLPEYPGVLSDIQEEKSIKYKFEVYEKNN

>DHFR_canFam Canis familiarus (dog) XM_003432480
MVRTLNCIVAVSQNMGIGRNGDLPWPPLRNEFKYFQRMTTNSSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRDLKEPPQGAHFLAKSLDDALKLTEQPELANKVDMVWIVGGSSVYKEAMNKPGHLRLFVTRIMHEFESDTFFPEIDLEKYKLLPEYPGVLSDVQEEKGIKYKFEVYEKND

>DHFR_vulVul Vulpes vulpes (fox) SRA|SRR094997.876315.2
MVRTLNCIVAVSQNMGIGRNGTVPWPPLRNEFKYFQRMTTNSSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRDLKEPPQGAHFLAKSLDDALKLTEQPELANKVDMVWIVGGSSVYKEAMNKPGHLRLFVTRIMHEFESDTFFPEIDLEKYKLLPEYPGVLSDVQEEKGIKYKFEVYEKND

>DHFR_musPut Mustela putorius (ferret) AEYP01048652/AEYP01048654/AEYP01048651/SRA|SRR071690.159842.2
MVRTLNCIAAVSQNMGIGKNGDLPWPPLRNEFKFFQRMTTTSSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINLVLSRDLKEPPQGAHFLAKSLDDALKLIEQPELANKVDMVWIVGGSSVYKEAMNKPGHLRLFVTRIMQEFESDTFFPEIDLEKYKLLPEYPGVLSDVQEEKGIKYKFEVYEknd

>DHFR_ailMel Ailuropoda melanoleuca (panda) XM_002916218
MVRTLNCIAAVSQNMGIGKNGDLPWPPLRNEFKFFQRMTTTSSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINLVLSRDLKEPPQGAHFLAKSLDDALKLIEQPELANKVDIVWIVGGSSVYKEAMNKPGHLRLFVTRIMQEFESDTFFPEIDLDKYKLLPEYPGVPSDVQEEKGIKYKFEVYEKND 

>DHFR_ursAme Ursus americanus (bear) GW292063 fragment
MVRTLNCIAAVSQNMGIGKNGDLPWPPLRNEFKFFQRMTTTSSVEGKQNLVIMGRKT

>DHFR_equCab Equus caballus (horse) XM_001504643
MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTASSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINLVLSRELKEPPQGAHFLAKSLDDALSLLEQPELANKVDMVWIVGGSSVYKEAMNKPGHLRLFVTRIMQEFESDTFFPEIDFEKYKLLPEYPGVLSDIQEEKGIKYKFEVYEKND

>DHFR_vicPac Vicugna paca (vicuna) wgsBlast
MVRPLDCIIAVSQNMDMGKNGDLPWPplrNEFKYFQRMTTASSAEGKQNLVIMGRKTWFSIPEKNRPLKDRINLVLSRELKEPPQGAHFLANSLDDALKVIEQPELTNKVDMVWIVGGSSVYKEAMDKPGHLRLFVTRIMQEFESDTFFPEIDLEKYKLLSEYPGVPSDVQEEKGIKYKFEVYEKNN

>DHFR_susScr Sus scrofa (pig) NM_001244064
MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEYKYfQRMTTTSSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPQGAHFLAKSLDDALKLTEQPELKNKVDMVWIVGGSSVYKEAMNKPGHIRLFVTRIMKEFESDTFFPEIDLEKYKLLSECSGVPSDVQEEKGIKYKFEVYEKNN

>DHFR_turTru Tursiops truncatus (dolphin) DT661408
MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEYRYFQRMTTTSSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPQGAHFLAKSLDGALKLIEQPELTNTVDMVWIVGGGSVYKEAMNKPGHLRLFVTRIMQEFESDTFFPEIDLEKYKLIPEYPGVPSDVQEEKGIKYKFEVYEKNN

>DHFR_oviAri Ovis aries (sheep) GO728717
MVRPLNCIVAVSQNMGIGKNGNLPWPPLRNEFKYFQRMTTVSSEEGKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPKGAHFLAKSLDDALELIEDPELTNKVDMVWIVGGSSVYKEAMNKPGHLRLFVTRIMQEFESDVFFPEIDFEKYKLLPEYPGVPLDVQEEKGIKYKFEVYEKNN

>DHFR_capHir Capra hircus (goat) JO612108
MVRPLNCIVAVSQNMGIGKNGNLPWPPLRNEFKYFQRMTTVSSEEGKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPKGAHFLAKSLDDALELIEDPELTNKVDMVWIVGGSSVYKEAMNKPGHLRLFVTRIMQEFESDVFFPEIDFEKYKLLPEYPGVPSDVQEEKGIKYKFEVYEKNN

>DHFR_bosTau Bos taurus (cow) NM_001077883
MVRPLNCIVAVSQNMGIGKNGNLPWPPLRNEFQYFQRMTTVSSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPKGAHFLAKSLDDALELIEDPELTNKVDVVWIVGGSSVYKEAMNKPGHVRLFVTRIMQEFESDAFFPEIDFEKYKLLPEYPGVPLDVQEEKGIKYKFEVYEKNN

>DHFR_myoLuc Myotis lucifugus (microbat) wgsBlast
MVRTLNCIAAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVEGKQNLVIMGRKTWFSIPKKNRPLKDRINLVLSRELKEPPKGAHFLAKSLDDALKLIEEPELTNQVDMVWIVGGSSVYKEAMSKPGHQRLFVTRIMQEFESDTFFPEIDLEKYKLLPEYPGVLSDVQEEKGIKYKFEVYEKNE
 
>DHFR_pteVam Pteropus vampyrus (macrobat) wgsBlast
MVLSLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSveGKRNLVIMGRKTWFSIPKKNRPLKDRINLVLSRELKEPPKGAHFLAKSLDDALKLIEQPELRSKVDMVWIVGGSSVYKEAMNKPGHIKLFVTKIMQEFESDIFFPEIDLGKYKLLPEYPGVLSDVQEEKGIKYKFEVYEKND

>DHFR_eriEur Erinaceus europaeus (hedgehog) wgsBlast/ti|909141827
MVRPLNCMVAVSQNMGIGKNGELPWPPLRNEFKHFRTMTSTPSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINLVLSRELKEPPQGAHFLAKSLDAALKLIDQPELTNKVDRV----------EVMNRPGHIRLFVTRILQEFECDTFFPEIDLKNYRLLP--------------------------

>DHFR_sorAra Sorex araneus (shrew) wgsBlast
MVRPLNCIVAVSQNMGIGKNGELPWPPLRNEFTYFRKMTTTSSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINLVLSRELKEPPTGAHFLAKSLDDALKLIEQPELTNKVDMVWIVGGSSVYQEVMNKPGHLRLFVTRIMQEFECDTFFPEIDLEKYKLLTDYPGVVSDVQEEKGIKYKFEVYEKKD

>DHFR_loxAfr Loxodonta africana (elephant) XM_003408117 error single trace TTTTTSSV
MVRPLNCIVAVSQNMGIGKNGDLP*PplRNEFKYFQRMTTTSSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINLVLSRGLKEPPQGAHFLAKSLDDALKLIEQPELRNKVDMVWIVGGSSVYKEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLEKYKRLSEYPGVLSDVQEEKGIKYKFEVYEKKD

>DHFR_proCap Procavia capensis (hyrax) wgsBlast
MVRPLSCIVAVSQNMGIGKNGDLPWPPLRNEFKHFRTMTSTPSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINLVLSRELKEPPQGAHFLAKSLDDALKLIEQPELTNKVDLVWIIGGSSVYKEAMSQPGHLTLFVTRIMQEFESDTFFPEIDLEKFKLLPEYPGVPSDVQEEKGIKYKFEVYEKKD

>DHFR_dasNov Dasypus novemcinctus (armadillo) AAGV020342833
mVRPLSCYDAVYQNMGIGKNGDMPWPPLRNEYKYFQKMTTVSSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELREPPQGAHFLAKSLDDALKLIEQPELADKVDMIWIVGGSSVYK---------------------------------------YPGVLSDVQEEKGIKYKFEVYEKND

>DHFR_choHof Choloepus hoffmanni (sloth) genome/blat many pseudogenes
-----------------------------NEFKYFQRMTTTSSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINLVLSRELK------------------------------------------EAMNRPGHLRLFVTRIMQEFESDTFFPEIDTEKYKLLPEYPGVLSDVQEEKGIKYKFEVYEKND

>DHFR_monDom Monodelphis domestica (opossum) XM_001363581
MVRQLNCIAAVSKNMGIGKDGDLPWPLLRNEFKYFQKMTTTPSVEGKQNLVIMGKKTWFSIPEKCRPLKDRINVVLSRELKEPPQGAHYLASSLDNALNLLELPELADKVDMVWIVGGSSVYKEAMNKPGHQRLFITRILQEFESDTFFPEIDLKKYKQLQDYPGVSPDVQEEKGIQYKFEVYEKHE

>DHFR_macEug Macropus eugenii (wallaby) wgsBlast
MAPTINCIVAVAQNMGMGKNGDLPWPPLRKEFQYFQKMTTTPSVEGKQNLLIMGKKTWFSIPEKNRPLKDRINLVLSRELKDPPLGAHYLAKSLDDALDLLRLPELANTVDMVWVVGGSSVYKEAMNKPGHQRLFVTRILQEFESDTFFPEIDLKRYKLLPEYPGVLTDVQEEKGIQHKCEIHEKHD

>DHFR_sarHar Sarcophilus harrisii (tasmanian_devil) AFEY01286966/AFEY01033989/AFEY01359230
MVRPINCIAAVSQNMGIGKNGDLPWPPLRNEFNYFQKMTTTSSVEGKQNLLIMGKKTWFSIPEKHRPLKDRINLVLSRELKEPPPGAHYLAKSLDDALDLLELPELADKVDMIWVIGGSSVYKEAMEKPGHQRLFVTRILQEFECDTFFPEIDLKKYKLLPEYPGVLTDIQEEKGIQYKFEVYEKHD

>DHFR_triVul Trichosurus vulpecula (brushtail) EG594753
MVRTINCIVAVSQNMGIGKNGDLPWPPLRNDFKHFQKMTTIPSVEGKQNLLITGKKTWFSIPEKSRPLKDRINVVLSRELKEPPPGAHYLAKSLDDALDLLELPELANKVDMIWVVGGSSVYKEAMNKRGHNRLFVTRILQEFESDTFFPEIDLKRYKLLXEYPGVLTDV

>DHFR_ornAna Ornithorhynchus anatinus (platypus) XM_001512186
MGPLLNCIVAVAKNGGIGNKGDLPWPALRNEFRYFQKMTTTPTVKGKQNLVIMGKKTWFSIPEKSRPLKDRINIVLSRELKEPPQGAHYVAKSLDDALNLLHLPELASKVDMVWIIGGSSVYKEAMEKGGHQRLFVTRILQDFESDTFFPEIDLEKYKLIPEYPGVPADTQEEKGIQYRFEVYEKHI

>DHFR_tacAcu Tachyglossus aculeatus (echidna) SRA:EUGXWLM02GHH9L/EUPZL4S02GL94B
MGRPLNCIAAVAKNGGIGNKGDLPWPPLRNEYRYFQKMTTTPTVEGKENLVIMGKKTWFSIPEKSRPLKDRINIVLSRELKEPPQGAHYVAKSLDDALNLLNLPELASKVDMVWIIGGSSVYKEAMEKPGHQRLFVTRILQEFESDTFFPEIDLEKYKLIPEYPGVPADTQEEKGIQYRFEVYEKHI

>DHFR_galGal Gallus gallus (chicken) NM_001006584 
MVRSLNSIVAVCQNMGIGKDGNLPWPPLRNEYKYFQRMTSTSHVEGKQNAVIMGKKTWFSIPEKNRPLKDRINIVLSRELKEAPKGAHYLSKSLDDALALLDSPELKSKVDMVWIVGGTAVYKAAMEKPINHRLFVTRILHEFESDTFFPEIDYKDFKLLTEYPGVPADIQEENGIQYKFEVYQKSVLAQ

>DHFR_melGal Meleagris gallopavo (turkey) genome/blat
------------QNMGIGKDGNLPWPPLRNEYKYFQRMTSTSHVE------------------------------------EAPKGAHYLSKSLDDALALLDSPELKSKVDMVWIVGGTSVYKAAMEKPINHRLFVTRILHEFESDTFFPEIDYKDFKLLKEYPGVPADVQEENGIQYKFEVYQKSVLAQ

>DHFR_lagLag Lagopus lagopus (ptarmigan) GW704257
MVRSLNSIVAVCQNMGIGKDGNLPWPPLRNEYKYFQRMTSTSHVEGKQNALIMGKKTWFSIPEKNRPLKDRINIVLSRELKEAPEGAHYLSKSLDDALALLDSPELKSKVDMVWIVGGTSVYKAAMEKPINHRLFVTRILHEFESDTFFPEIDYKDFKLLTEYPGVPADVQEENGIQYKFEVYQKSVLAQ

>DHFR_anaPla Anas platyrhynchos (duck) scaffold675
MVRSLNSIVAVCQNMGIGKDGNLPWPPLRNEYKYFQRMTSTSHVEGKQNALIMGKKTWFSIPEKNRPLKDRINIVLSRELKEAPEGAHYLSKSLDDALALLDSPELKSKVDMVWIVGGTSVYKAAMEKPINHRLFVTRILHEFESDTFFPEIDYKDFKLLTEYPGVPADVQEENGIQYKFEVYEKTVLAQ

>DHFR_taegut Taeniopygia guttata (finch) XM_002190440
MPRSLNSIVAISQNMGIGKDGRLPWPPLRNDYKYFQRMTSASPVEGKKNAVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEAPKGAHYLSKSLDDALALLDSPELQSKVCMVWIVGGSSVYKEAMEKPIHHRLFVTRILKEFESDTFFPEIDHKDYKLLTEYPGVPADIQEENGIQYKFEVYEKTVVP

>DHFR_ficHyp Ficedula hypoleuca (flycatcher) SRA|SRR029160.210264.2
MVRSLNSIVAVSQNMGIGKDGRLPWPPLRNEYKYFQRMTSTPRVEGKQNALIMGKKTWFSIPEKNRPLKDRINIVLSRELKEAPEGAHYLSKSLDDALALLDSPELKSKVDMVWIVGGTSVYKAAMEKPIHHRLFVTRILKEFESDTFFPEIDHKDYKLLTEYPGVPADIQEENGIQYKFEVYEKAVV*

>DHFR_melUnd Melopsittacus undulatus (budgerigar) SRA|SRR029329.119695.2
MVRSLNSIVAVCQNMGIGKDGSLPWPPFRNEYKYFQRMTSTSQVEGKQNALIMGKKTWFSIPEKNRPLKDRINIVLSRELQETPKGAHYLSKNLDDALALLDSPELKSKVDMVWIVGGTSVYKAAMEKPINHRLFVTRILQEFESDTF

>DHFR_allMis Alligator mississippiensis (alligator) genome/blat 
MVSSLNAIAAVSQNMGIGKNGTLPWPPLRNEFKYFQRMTTTPSVPGKQNVLIMGKKTWFSIPEKNRPLKDRINIVLSRELAEVPKGAHHLARSLDDALVLLDSPELKNKIDLVWIVGGSSVYKAAMEKPINQRLFVTRILHEFESDTFFPEIDLEKYRLLPEFPGVPADVQEENGIHYKFEVYEKT

>DHFR_croPor Crocodylus porosus (crocodile) genome/blat composite seq with alligator
MVSSLNAIAAVSQNMGIGKNGTLPWPPLRNEFKYFQRMTTTPSVPGKQNVLIMGKKTWFSIPEKNRPLKDRINIVLSRELAEVPKGAHHLARSLDDALVLLDSPELKNKIDLVWIVGGSSVYKAAMEKPINQRLFVTRILHEFESDTFFPEIDLEKYRLLPEFPGVPADVQEENGIHYKFEVYEKTV

>DHFR_chrPic Chrysemys picta bellii (turtle) AHGY01047456 AHGY01047458
MVRPLNCIAAVCQNMGIGKNGDLPWPPLRNEFKYFQRMTTTTTVEGKQNVLIMGKRTWFSIPEKNRPLKDRINIVLSRELKETPKGAHYLAKSLDDALVLLDSPELKSKIDMVWIIGGSSIYKAAMEKPVHQRLFVTRILHEFESDTFFPEIDLEKYKHLPEYPGVPADVQEENGIRYKYEVYEKTV*

>DHFR_anoCar Anolis carolinensis (lizard) genome/blat XM_003216272
MVLSLNSIAAVCQNMGIGKNGQLPWPPLR 2NEFKYFQKMTMTPTQEGKQNVVIMGKRTWFSIPEKNRPLKDRINIILSREMKEVPKGAHYLARSLDEALDLLQSPELENKIDLVWIVGGSSVYK AAMDKPIHHRLFVTRILHDFESDTFFPEIDLEKYKLIPSYAGVPTDIQEENGIRYRFEVYEKNI

>DHFR_pytMol Python molurus (python) AEQU011279324/AEQU010134262/AEQU010204446/AEQU010142865/AEQU010435622/AEQU010656196
MVASLHSIVAVCNNMGIGKDGKLPWPPLRNEFKHFQKMTMTTKEEGKQNVVIMGKKTWFSIPEKHRPLKNRINVVLSKELKDVPEGAHYLAKSLEEALDLLETSEMERKVDKVWIVGGSSVYK AAMEKPIHHQLFVTRIMHDFESDTFFPEIDLKKYKLLPKYTGIPTDIQEENGIQYKFEVYENTI

>DHFR_ambMex Ambystoma mexicanum (axolotl) CN050864
MGRILNAIVAVCPSMGIGKDGNLPWPILRNEFKYFQRMTMTATQEGKQNVVIMGRKTWLSIPEKNRPLKGRINIVLSRELTAPPPGAHYLANSLDGALELLETRELAGKVDLLWIIGGSSLYRELIGRPHHQRLFVTRILKEFECDTFLPEFDLEKYRLLPEYPGVSLGIQEENGIQYKFEVYEKVEEA

>DHFR_xenTro Xenopus tropicalis (frog) CT025223
MRNPFLHAVVAVCPPNQGIGKEGSLPWPLLRNEFKHFQRLTMTPTVEDKKNVVIMGRKTWFSIPEKNRPLKERINIVLSKELKEPPTGAHYLSKSLAEAIDLLEDPELKDKVDLVWVIGGSSLYQELMEKPVNQRWFVTRILQEFECDTYLPEIDLNSFRLLPEYPGISPELQEENGVQYKFEVYEKIV

>DHFR_xenLae Xenopus laevis (frog) NM_001095037
MRNQFLHAVVAVCPPNQGIGKGGSLPWPLLRNEFKHFQRLTMTPTVEGKKNVVIMGRKTWFSIPEKNRPLKERINIVLSKELKEPPTGAHYLSKSLTEAIDLLEDPELKDKVDLVWIIGGSSLYQELMEKPINQRWFVTRILQEFECDTYLPEINLNSFQLLPQYPGISPELQEENGVQYKFEVYEKIV

>DHFR_ranCat Rana catesbeiana (bullfrog) GO463031
---------------------GGTLPWPLLRNEFKHFQRLTMTPTTEGKQNVVIMGRRTWFSIPEKNRPLKGRINIVLSRELTEPPNGAHYLAKSLDEAVHLLKTPEMKEKIDLIWIIGGSSLYKEVMDKPVSQRLFITRIQQEFECDTFLPDVDLQSYKLLPEYPGISSEMQEENGIQYKFEVYEKML

>DHFR_latCha Latimeria chalumnae (coelocanth) AFYH01150805/AFYH01150806/AFYH01150807/AFYH01150815/AFYH01150814
MGAARLLNSIVAVCPNLGIGKDGNLPWHPKRLSNEFRYFQKMTTTPTVEGKQNVVIMGRKTWFSIPEKNRPLKGRVNVVLSRELQEAPEGAHYLARDLDSALKLLESPELADRVDLVWIIGGSALYKEALEHPSQHRLFVTRVLQQFECDTYLPEINFNKFTLLPEFPGVSSAIQEESGIQYKFEVYEKNS

>DHFR_lepOcu Lepisosteus oculatus (spotted_gar) AHAT01012362
0 MPRPINCIVAVCPNMGIGHNGNLPWHPKRL 2
1 NEFKYFQKMTMTPTLE 1
2 GQQNAVIMGRKTWFSIPERNRPLKNRINIVLSRELK 2
1 NPPEGAHYLASDFSSALRLLDSAPQTDQVDQVWVIGGSSVYK 0
0 EAMEAPGWCRLFVTQILQDFECDTFLPEINLNKYRLLPE 2
1 FPGVPLEVQEENGIRYKFEVYESIQN* 0

>DHFR_gadMor Gadus morhua (cod) FF410604 novel insertion 3 transcripts
MSRVLNCIVAVCPDAGIGYKGDLPWHPTRLNNEFKHFRRLTVTrtPGAEDKQNVVIMGRKTWNSIPEKNRPLNDRINIVLSRQLEAPPSGAHHLARDLPSALRLLDVPEGLLADQVWLIGGSSMYSEVMETPCPRRLFVTRILKQFPSDTFFPDISPAQFRLLPEYPGVSSELQEENGIQYRFQVYQSVDP

>DHFR_tetNig Tetraodon nigroviridis (pufferfish) CR710339
MARVLNAIVAVCPDLGIGRNGDLPWHPIRLDNEFKHFRKMTSTPSVNGKQNVVIMGRKTWFSIPEKHRPLANRINIVLSRRSREPPAGAHHLAHDLSSALQLVETQLAEQADQVWVIGGSSLYQEMMERPGTSRLFVTHVLKQFDCDTFLPEIRPQRYRLLPQFPGVPQELQEEKGIQYRYQVYESLEP

>DHFR_hipHip Hippoglossus hippoglossus (halibut) EB031730 
MSRILNGIVAVCPDLGIGNRGNLPWHPVRLSNEFKHFRSMTATPSEKGKQNVVIMGRKTWFSIPEKNRPLNNRINIVLSRQCKAPPAGAHHLSADFSSALRLVDTELADQADQVWVIGGSSLYKEMMESPGTRRLFVTRILKQFDCDTFLPEISPDKYRQLPEFPGVPQELREENGIQYRFEVYESIEP

>DHFR_solSen Solea senegalensis (sole) FF286406 
MSRVLNGIVAVCPDMGIGMTGNLPWHPVRLSNEFKHFRTMTATSSVKGKKNVVIMGRKTWFSIPEKNRPLNNRINIVLSRQCKLPPAGAHHLAADFSSALRLVETELADEADQIWIIGGSSLYKEMMESPGIKRLFVTRILKQFECDTFLPEISLDKYRLLPEFAGVSQELQEENAIQYRFEVYESVEQ

>DHFR_oreNil Oreochromis niloticus (tilapia) GR620119
MPRVLNAIVAVCPDRGIGNKGNLPWHPIRLSKEFAHFRKMTATPSVKGKQNVVIMGKKTWYSIPEKNRPLSNRINIVLSRECKVPPAGAHYLASDFSSALRLIDTELADQADQVWVIGGSSLYKEMMGSTGMRRLFVTQILKQFECDTFLPEISLDKYRLLPEFPDVPQELQEENGIQYRFKVYESIQE

>DHFR_dicLab Dicentrarchus labrax (seabass) FM022259 
mSRILNGIVAVCPDLGIGMNGNLPWHPVRLGNEFKHFRTMTATPSVKGKQNVVIMGRKTWFSIPEKNRPLNNRINIVLSKECKVPPAGAHHLAADFNSALRLVETELADQADQVWVIGGSSLYKELMESPGTRRLFVTRILKQFECDTFLPEICPDKYQLLPEFPGVSQELQEENGIQYRFEVYESTEQ

>DHFR_perFla Perca flavescens (perch) GO655828 
MSRILNGIVAVCPDLGIGNNGNLPWHPVRLNNEFKHFRTMTATPSVNGKQNVVIMGRKTWFSIPEKNRPLNNRINIVLSRECKAPPAGAHHLARDFSSALRLVDTELAQQADTVWVIGGSSLYKEMMESPGTRRLFVTRILKQFECDTFLPEITPDKYRLLPEFPGVPQELQEENGIQYRFEVYESIQQ

>DHFR_spaAur Sparus aurata (seabream) FM155123 
MSRIVNGIVAVCPDLGIGNNGNLPWHPVRLNREFKHFRTMTATPSVKGKQNVVIMGRKTWFSIPDKFRPLNNRINIVLSRECKVPPAGAHHLAADFSSALSLVDTELADQADQVWVIGGSSLYKELMESTRTSRLFVTRILKQFECDTFLPEISPDKYSLLPEFPGVPQGLQEENGIQYKFEVYERIKQ

>DHFR_gasAcu Gasterosteus aculeatus (stickleback) ENSGACT00000004939 
MSRVLNGIVAVCPDLGIGCHGNLPWHPLRLSNEFKHFRRMTATASVKDKQNVVIMGRKTWFSIPEKNRPLNNRINIVLSRELKAPPAGAHHLAPDFSSALRLVDTELAEQADQVWVIGGSSLYKELLESPGTRRLFVTRILKQFECDTFLPEICPDRYRLLPEFPGVPEELQEENGVQYRFEVYESIDV

>DHFR_oryLat Oryzias latipes (medaka) BJ496706
MTRTLNGIVAVCPDLGIGKGGNLPWHPLRLSKDFALFRKMTSTPLVAGRQNVVIMGRKTWFSIPEKNRPLQNRINIVLSRQLKAPPAGAHFVAADFAAALRLVDSELAERADQVWIIGGSSIYXEMMESPAPRRLIVTRVQKQFECDTFFPEIRPEQFRLLPEFPGVSQELQEENGVSYKFEVYESIQQ

>DHFR_anoFim Anoplopoma fimbria (sablefish) BT082890
MSRVLNAIVAVCPDLGIGRNGDLPWHPVRLNNEFKHFRRMTSTPSVEGKQNVVIMGRKTWFSIPEKNRPLNNRINIVLSRELKTPPAGAHHLAQDFNSALRLVDMKLAEQVDQVWVIGGSSLYKELMESQGIRRLFVTQILKQFECDTFLPKISPDKYRLLPEFPGVPQELQEENGIQYRFEVYESIEQ

>DHFR_esoLuc Esox lucius (pike) BT079933
MSRVLNCIVAVCPNMGIGNKGNLPWHPKRLNNEFKYFQKMTMTPFVEGKQNVDIMGRKTWFSIPERNRPLKNRINIVLSRELKEPPPGAHHLASDFSSALRLLDTAEMAERVDQVWVIGGSSLYKEMMESEGIRRLFVTRILQQFESDTFLPEINQDKYRLLPEFPGVPQDVQEESGIQYRFEVYESVDH

>DHFR_salSal Salmo salar (salmon) BT047529
MSRVPNCIVAVCPDVGIGNNGNLPWHPKRLNNEFKYFQKMTMTSSVEGKQNVVIMGRKTWFSIPERNRPLKNRINIVLSRELKKPPAGAHHLASDFRSALCLLDMAEMAERADQVWVIGGSSLYKEMMESQGTRRLFVTRILQQFKSDTFLPEINQDQYRLLPEFPGVPQDLQEEAGLQYRFEVYESIDQ

>DHFR_oncMyk Oncorhynchus mykiss (trout) EZ786723
MSRVLNCIVAVCPDMGIGNNGNLPWHPKRLNNEFKYFQKMTMTSSVEGKQNAVIMGRKTWFSIPERNRPLKNRINIVLSRELKEPPAGAHQLASDFRSALRLLDTAEMAERVDQVWVIGGSSLYEEMMESQGTRRLFVTRILQQFKSDTFLPEINQDKYRLLPEFPGVPQDLQEEAGLQYRFEVYESIDQ

>DHFR_danRer Danio rerio (zebrafish) NM_131775
MSRILNCIVAVCPDMGIGKNGNLPWHPIRLSNELKHFQKMTMTPSDEGKKNVVIMGRKTWFSIPAAHRPLKNRINIVLSRELKTAPEGAHYLASDFSSALHLLDSGELEKLVDQVWIIGGSSLYKEVMERSGHRRLFVTRILKQFDCDTFIPNFDMDKYKLLPEFPGVPVGLQEDNGVQYLFEVYESIKH

>DHFR_cteIde Ctenopharyngodon idella (carp) CK233011 
MSRILNCIVAVCPDMGIGRKGNLPWHPIRLSNEFKHFQKMTMTPSVEGKKNVVIMGRKTWFSIPAQNRPLKNRINIVLSRELKTAPEGAHYLASDLSSALHLLDSSELEKQVDQVWIIGGSALYKEVMERSGPRRLFVTRVLKQFDCDTFIPEINMDKYKLLPEFPGVPTGLQDENGVQYVFECMIVS

>DHFR_cypCar Cyprinus carpio (carp) CA968873 
MSRILNSIVAVCPDMGIGKNGNLPWHPIRLSNEFKHFQKMTMTPLVEGKKNVVLMGRKTWFSIPAANRPLKNRINIVLSRELKTAPEGAHYLASDFSSALHLLDSSELEKQVDQVWIIGGSALYKEVMESSGPRRLFVTRVLKQFDCDTFIPDINMDKYKLLPEFPGVPTGLQEENGVQYVFEVYESTDH

>DHFR_ictPun Ictalurus punctatus (catfish) FD330522 
MGRVLNCIVAVCPDMGIGRNGNLPWHPIRLSKEFKHFQKMTMTPTVEGKKNVVIMGRKTWFSIPAQNRPLKNRINVVLSRELKTAPEGAHYLAADFSSALRILENAELEAQADQVWIIGGSSLYKEVMESSAHQRLFVTRILKQFDCDTFIPNIDLDKFRLLPEFPGVPAGLQEENGLQYVFEVYESVEQ

>DHFR_leuEri Leucoraja erinacea (skate) EE989413 
MTRLINSIVAVCPNMGIGNNGNFPWHPIRLSKEFKHFQRMTSTPSVEGKCNAVIMGRKTWHSIPEKNRPLKNRINIVLSRELKEVPEGAHYLAPDLDSALALLDSSDLQDKVDLVWIIGGSALYKEALESPASHRLFVTRILQDFDSDTFIPEINLNKFKLLPEFPDVPADIQEENGIRFKYEVYERISPV

>DHFR_squAca Squalus acanthias (dogfish) DV497998 
mPRLVNCIVAVCPNMGIGKDGNFPWHPIRLSKELKHFQKMTATPSVEGKRNAVIMGRKTWFSIPEKHRPLKDRLNIVLSRELKELPHGANYLAHDLDSALAHLDSSELQDKVDLVWIIGGSTLYKEAMESPVSHRIFVTRILKDFESDTFLPEMDLNKFKLLPKFPDVPTEVQEDNGIQYKFEVYERIAHA

>DHFR_eptBur Eptatretus burgeri (hagfish) BJ646201
MAQNPVNVIAAVLPNMGIGWKGGLPWHSKSLVKEMKHFTRLTSAAAEGKQNAVVMGRKTWESIPEKFRPLRNRINVVLSRTLSHLPLGAHYLATDFSNALAQLSESELCCRIDHIWIIGGSSIYKEALLHPACHRIFITRIMHDFEADTFFPHIDSKRFSLCQSYPGIPEEIQEENGVQYKFEVYEVVAK

>DHFR_petMar Petromyzon marinus (lamprey) Contig6033:1-27,714
GKQNVVIMGRKTWFSIPEKKRPLKNRINIVLSRELKCLPEGAHYLVPDLNSALSLIDSTPAIQEKLDGVWILGGGGVYKEAMEHSACRRLFITRVLQTMEADAFFPDIDADKFKLLP

Urochordate, cephalochordate, hemichordate and echinoderm reference sequences

>DHFR_cioInt Ciona intestinalis (tunicate) F971791 dozens of transcripts
0 MPAKDIQIHSVVACCNNGGIGFKGRLPWRLP 2
1 KEMKYFKRITTGEVEEFG GRRNaiIIGRKTWESIPKSFKPLKDRINIVISRTLDKDCEGP 0
0 DLVVNSLDELIdLLHSKPWCDKINQVFNIGGNEIYK 0
0 LIHNSQYCGKIYLTRVLADlQCDTFLENLDDNFTKLPTEG 2
1 FPEVPQGIQMDKNDLQWKVEVYEKKTT* 0

>DHFR_cioSav Ciona savignyi reftig_155
0 MPAKELKIHSIVACCNNRGIGNKGRLPWRLP 2
1 KEMKHFTSLTTGDVENLN GKRNAVVVGRKTWESIPKSFKPLKNRVNIVISRSLSKEYEGP 0
0 DAVVNSLEELVQLLQSDHWSNRINQVFNIGGSEVYK 0
0 LINNSKYCGTIYLTRVLADFECDTFLDKFDDTFKQLDTTK 2
1 FPEVPQGEQMDKNDISWRVEVYVKKS* 0

>DHFR_halRor Halocynthia roretzi (squirt) DB618016/EY457317
0 MKIHAVVACCNNRGIGKDGTLPWRLP 2
1 KEMAHFQKITKGSPPK  GKKNAVVMGRKTWEEIK   RPLKDRYNIVMSRSLSEKLKGM 0
0 DALVRSLGELEDLLSSGEWTGEINEVFNIGGAEICR 0
0 LINGSSLCGRLCVTQMVTDFECDAFLPEIDDGEYLR LDE 2
1 YPEDVATLLPDGEVVEANGVKWAVEVYEAI* 0

>DHFR_botSch Botryllus schlosseri (tunicate) HO068783
0 MEIHTVVVCDDNWGISKDGCLPFGTK 2
1 KEAAHFYRFIADTSHLPP GRKNVLIMGRKTWTE ETPADAPQSRSVTYNFLLSKTLKERPEEV 0
0 DGIVSSVEEFVTLMESPRWKDKIGRIYNIGGGQACE 0
0 AIYASKYCGRAYMSLVRGDFHCDTFAPNLD

>DHFR_molTec Molgula tectiformis CJ399083 poor quality fragment
MGRNTWNSIPSKFRPlKnRWNIVLSRTlKDPyDGM
HAVVSSlDQlYKlLQDEAWKDKVNEIFCIGGAQVYE
VIDNSPYIGLiFMTRIRKKYDCDTFYSEVNKDKFILLKSCP

>DHFR_oikDio Oikopleura dioica (tunicate) CABV01002330/FP801526 completely different intronation from Ciona
0 MNKSGWNMILAADIKGGIGLRNDLPWRIPQDLKHFQMLTKGTPNQESVVVMGRNTWQSIPEKFRPLK 0
0 GRVNVVITNTLESENFIKCTSLEAAIAYINENHVNATKWIIGGAtLYNRGLKENFIDEVYLTRIQGDY 1
2 NCDTYVKDLYKILEVDYRLETSPKFDSKEVIGESGIKYAFERYVKNSGI* 0

>DHFR_braFlo Branchiostoma floridae (amphioxus) ABEP02022512/XM_002597864/BW702700 triplicated contig misassembly
0 MKTKKLSLVVAACNNMGIGVDGKIPWTLR 2
1 GDMKFFSRLTSGTEEA 1
2 GKQNAVVMGRKTWFSIPDRFRPLPKRLNVVLSRNLT 2
1 TPPEGAHHLAGSLEEAVKMLTETTVADTIDKVFIIGGNSVYK 0
0 DALSHPCCHRVYLTRVYKDFHCDTFFPNMDDTFKLVS 2
1 DPTIPSEMQEENGIEYKYEVYEKVA* 0

>DHFR_sacKow Saccoglossus kowalevskii (acornworm) ACQM01022605 exons 2-3 fused
0 MKKISLVAAACNNMGIGKNGDLPWRLR 2
1 KEMSFFTKVTSETKED GKQNAVVMGRKTWFSIPEKYRPLAGRYNVILSKNLK 2
1 ECPSGADMLSESLDDAIKQLSEPPLVDKIDKVYIIGGSSVYK 0
0 ESLESAACYRIYLTRVMADFDCDTFFPEFDTNLFNLVS 2
1 DPDVPSEIQEEKGIQFKYEVYEKK* 0

>DHFR_balCla Balanoglossus clavigerus (acornworm) FN986971
mQKISPVAAACNSMGIGKNGNLPWRLRKEMKYFTNVTSETVEEEKQNAVIMGRKTWFSIPEKYRPLNNRFNVILSNTLKECPAGADLLSPSLEEAVKLLSESPLEGKIDKVFIIGGSSVYKESIQSSLCHRIYLTRIMADFDCDTFFPNFDTNIFKLISDPSIPSEIQEEKGIQFKYEVYERQDV

>DHFR_strPur Strongylocentrotus purpuratus (urchin) AAGJ04143548  exons 2-3 fused
0 MAEKKLNLIAAACTSkGKMGIGINGNLPWRLR 2
1 QEMAYFERLTKTAQME GMKNAVIMGRKTWDSIPEKFRPLKDRVNVVLSNSLT 2
1 ECPPGADHLCSSLNEAVKLFSSPPLSETVDMVWITGGSAVYK 0
0 DGIDSPHCHRIYLTRIMKEIECDTFFPEFDLDRFKLVT 2
1 DPAVDQDTQEEKGIQYKFEIYESSS* 0

>DHFR_parLiv Paracentrotus lividus (urchin) AM567148
MADKRLNLIAAACTSNGKMGIGINGNLPWRLRQEMAYFERLTKTSQMEGMKNAVVMGRKTWFSIPEKFRPLKDRVNVVLSSSLTECPPGADHLCSSLSEAVKLLSSPPLSQTVDMVWITGGSSVYKDGIDSPLCHRIYLTRIMKEIECDTFFPEFDLDRFKLVTDDAVSQDIQEEKGIQYKFEIYESS

>DHFR_lytVar Lytechinus variegatus (urchin) JI431194
MAEKKLNLIAAACTSSGKMGIGINGNLPWRLRQEMAYFERLTKTPQLEGMKNAVIMGRKTWFSIPEKFRPLKDRINVVLSNTLTECPSGADHLCSSVSEAVKLLSCPPHSQTVDMLWIIGGSAVYKAGIESPCCHRIYLTRVLKEIECDTFFPEFDLEKFKLVVDPAVSQDIQEEKGIQYKFEIYENSS

>DHFR_patPec Patiria pectinifera (starfish) DB389444/DB385866 also called Asterina
MAGQKQcNLIVAACKCKDSLGIGINGTIPWKLRTDMKFFSTQTSTTAEN DKKNAVIMGRKTWLSIPDKFRPLPNRVNIVLSKTLSECPAGADHLCRSLDHAVKLLSEPPVADGIDVVWIIGGSSVYKEAMESSLCHRIYLTRIHSQFECDTFMPEIDTSKFKVVSDPAVSGERQKENDIEFNFEIHQNIQHTSTS

Protostome reference sequences

>DHFR_droMel Drosophila melanogaster (fruitfly) NM_001043255 Diptera; Muscomorpha 2-6 fused 
MLRFNLIVAVCENFGIGIRGDLPWRIKSELKYFSRTTKRTSDPTKQNAVVMGRKTYFGVPESKRPLPDRLNIVLSTTLQESDLPKGVLLCPNLETAMKILEEQNEVENIWIVGGSGVYEEAMASPRCHRLYITKIMQKFDCDTFFPAIPDSFREVAPDSDMPLGVQEENGIKFEYKILEKHS

>DHFR_droSim Drosophila simulans (fruitfly) XM_002103136 Diptera; Muscomorpha 2-6 fused
0 MLRFNLIVAVCENFGIGIKGDLPWRIK 2
1 SELKYFSRTTKRTSDPTKHNAVVMGRKTYFGVPESKRPLPDRLNIVLSTTLQESDLPKGVLLCPNLETAMKILEEQNEVENIWIVGGSGVYEEAMASPRCHRLYITKIMQKFDCDTFFPAIPDSFREVAPDSDMPLGVQEENGIKFEYKVLEKQS
	
>DHFR_droSec Drosophila sechellia (fruitfly) XM_002041171 Diptera; Muscomorpha 2-6 fused
0 MLRFNLIVAVCENFGIGIKGDLPWRIK 2
1 SELKYFSRTTKRTSDPTKHNAVVMGRKTYFGVPESKRPLPDRLNIVLSTTLQESDLPKGVLLCPNLETAMKILQEQNEVENIWIVGGSGVYEEAMASPRCHRLYITKIMQKFDCDTFFPAIPDSFREVAPDSDMPLGVQEENGIKFEYKVLEKQS
	
>DHFR_droEre Drosophila erecta (fruitfly) XM_001980159 Diptera; Muscomorpha 2-6 fused
0 MIRFNLIVAVGENFGIGYKGDLPWRIK 2
1 SELKYFSRITKRTNDPSKHNAVVMGRKTYFGVPESKRPLSNRLNVVLSTTLQENELPEGVLLCSNLETALKVLENKNDVENVWIVGGSGVYKEAMASPRCHRLYITKILHKFECDTFFPAIPDSFREVAPDSDIPLGVQEENGIHFEYKVLEKQSK

>DHFR_droYak Drosophila yakuba (fruitfly) XM_002097560 Diptera; Muscomorpha 2-6 fused
0 MLSFNLIVAVCENFGIGINGNLPWRIR 2
1 SELKYFSRTTKRTSDPSKHNAVVMGRKTYFGVPEGKRPLPDRLNIVLSTTLQENELPKGVLLCRNLEAAMKALEDKNEVENIWIVGGSGVYEEAMASTRCHRLYITKIHEKFECDTFFPAIPDSFREVAPDSDTPLGLQEENGIKFEYKVLEKQ

>DHFR_droAna Drosophila ananassae (fruitfly) XM_001954865 Diptera; Muscomorpha 2-6 fused
0 MRRFSLIVAVSENLGIGIKGDLPWRIK 2
1 SELKYFSRTTKRTSDPSKRNAVIMGRKTYFGVPESKRPLPNRLNIVLSTTLQPEDLPKDVMLCPNLEAAMKHLEGDDQIENVWVVGGSGVYAEAMDSNRCHRLYITRIHQKFECDTFFPTIPESFREVAPDPDTPAGVQEENGIEYEYKVMEKETK

>DHFR_droPse Drosophila pseudoobscura (fruitfly) XM_001359349 Diptera; Muscomorpha 2-6 fused
0 MLRFNLIVAVSENFGIGLKGDLPWRLK 2
1 SELKYFSRTTKRTSDPNKRNVVIMGRKTYFGVPASKRPLPERLNIVLSTTLKPAELPSDVVLCHSLEEAMKHLEEETLRDQIENVWIVGGSGVYEEAMASSRCNRLYITKIQQNFECDTFFPSIPDSFREVPPDADTPVGVQEENGIKYEYKVLEKRE
	
>DHFR_droPer Drosophila persimilis (fruitfly) XM_002017170 Diptera; Muscomorpha 2-6 fused
MLRFNLIVAVSENFGIGLKGDLPWRLK 2
1 SELKYFSRTTKRTSDPNKRNVVIMGRKTYFGVPASKRPLPERLNIVLSTTLQPAELPSDVVLCPSLEEAMKHLEEETLRDQIENVWIVGGSGVYEEAMASSRCNRLYITKIQQNFECDTFFPSIPDCFREVPPDADTPVGVQEENGIKYEYKVLEKRE

>DHFR_droWil Drosophila willistoni (fruitfly) XM_002070058 Diptera; Muscomorpha 2-6 fused
0 MLRFNLIVAVCENSGIGLKGDLPWRIR 2
1 SELKYFSNTTKRRLDSSKRNVVIMGRKTYFGVPPSKRPLPDRVNVILSSTLKSNELPNEVLIYSSLEGAMQELEKQKDQIENVWIVGGSSVYAESMASPRCHRLYLTKIYAEFECDTFFPKIPESFREVSPDKDTPPGIQEENGLKYEYKVFEKQE

>DHFR_droMoj Drosophila mojavensis (fruitfly) XM_001998634 Diptera; Muscomorpha 2-6 fused
0 MLKYNLIVAVCENFGIGFKGDLPWRLK 2
1 SELKYFSRRTKRINDPSKRNVVIMGRKTYFGVPPSKRPLPDRLNIVLSTTLTQKELPEGVLLCGSLDEAMQTLEKKPLCDQVENIWIVGGSGVYAEAMESPRCHRLYITKIQERFECDTFFPHIPASFHEVELTEDTPHGVQEENGIKFEYKILEKREHII

>DHFR_droVir Drosophila virilis (fruitfly) XM_002056347 Diptera; Muscomorpha 2-6 fused
0 MLRYNLIVAVCENFGIGLKGDLPWRLK 2
1 SELKYFSRTTKRTRDPAKRNVVIMGRKTYFGVPPSKRPLPDRLNIVLSTTMSRAELPNDVLLCRSLEEAMKMLESESEPWCDQIENIWIVGGSGVYEEAMASPRCYRLYITKIQEQFECDTFFPNIPADFHEVPLDEDTPHGVQEENGIKYEYKILEKQ

>DHFR_droGri Drosophila grimshawi (fruitfly) XM_001990307 Diptera; Muscomorpha 2-6 fused
0 MLKFNLIVAVCENFGIGFKGNLPWNLK 2
1 SELKYFSRTTKRVCDPSKRNVVIMGRNTYMGVPPSKRPLPDRLNIVLSTTMSAADLPKEVLLCRSLNEAMETIESPEWCKQVENVWIVGGSRVYAEAMDSERFHRLYLTKIHEKFECDTFFPEIPDCFKEIPLDEHTPDGIQEEDGIKYEYKILEKQ

>DHFR_gloMor Glossina morsitans (tsetse_fly) EZ423753 Diptera; Muscomorpha
MLKFNLIVAVSKNFGIGLKGGLPWELKSELRYFSELTKRVFDSTKRNVVIMGRKTYFGIPLNNRPLRNRLNIVLSTTLNKVGELPEEVLLQPNLEAAMKFLEDNNTLKSNIENIWIIGGASVFKEAMASKRCHRLYITEIQSSFESDVFLPTIPNDFEQIIPGPEVPQTVQVENCICFRYKVLEKRE

>DHFR_haeIrr Haematobia irritans (buffalo fly) AY099474 Diptera; Muscomorpha
MLKFSLIVAVCENFGIGIKGDLPWLLKSELKYFSTTTKRVHDPSKRNVVIMGRKTYFGVPESKRPLQQRLNIVLSTTLKQSDLPKDVLLYPNLESAMSFLENEKDINQQIETVWIVGGSGVYKEAMASPRCHRLYLTKIYAKFDCDTFFPDIPADFKEVDLDPETPKGIQEENNITYEYKILEKQDHK

>DHFR_sarCra Sarcophaga crassipalpis (flesh_fly) EZ609788 Diptera; Muscomorpha
MLKFSLIVAVCENFGIGIKGDLPWKIKSELKYFSSTTKRVRDPSKRNVAIMGRKTYFGIPESKRPLPERLNIVLSTTLKPSDLPSDVLLYPNLESAMKMLEENEKLKQEIETVWIVGGSGVYAEAMASQRCHRLYLTKIQASFECDTFFPKIPDDFVEVELDEETPKGIQEENNIKYEYKILEKKT

>DHFR_culQui Culex quinquefasciatus (mosquito) XM_001864570 Diptera; Nematocera exons 2-6 fused
0 MKKFSLIVAVCSNGGIGIKGDLPWRLR 2
1 SELRHFARMTKRVADPGKRNAIIMGRKTYFGIPEGRRPLPDRLNVVLTRDPASNTYPEGVLVCTSLEDALAKIQDLVENIWIIGGSAVYKEAMESRHCHRIYLTEILAPFECDAFFPEIGKEFRLVGNDADLAEEVQEENGVRFQYKIYEKKDID

>DHFR_anoGam Anopheles gambiae (mosquito) XM_319471 Diptera; Nematocera exons 2-6 fused
0 MSKKFSCIVAVCENRGIGINGDLPWKLK 2
1 QELKYFSHTTKKVNDADKRNAVIMGRKTYFGVPESKRPLPERLNIVLTRNAAAYTFPPDVLVCGSLQEALHKLDSTDHGEHIENIWVVGGNSVYKEAMESDRCHRIYLTEIKKQFDCDAFFPEIPNSFMVIDNDPDTPSGVQEENGIQYVYKIYENKQQ

>DHFR_aedAlb Aedes albopictus (mosquito) X60192 Diptera; Nematocera exons 2-6 fused
0 MKKFSLIVAVCANGGIGIKGDLPWRLR 2
1 QELKYFSRMTKKIQDSGKRNAIIMGRKTYFGVPESKRPLPERLNIILTRDPSANAYPSEVMVCTSMQEALKKLDEAPLVNEIENVWIVGGNAVYKEAMQSDRCHRIYLTEIKETFECDAFFPEITSDFQLVKNDDDVPEDIQEENGIQYQYRIYEKVPK

>DHFR_aedAeg Aedes aegypti (mosquito) AY432005 Diptera; Nematocera exons 2-6 fused
MKKFSLIVAVCANGGIGIKGDLPWRLR 2
1 QELKYFSRMTKKITDTSKRNAIIMGRKTYFGVPESKGPLPERLNIILTRDPSANTYPSDVMVCKSMQEALTKLDESPLANEIEKIWIVGGNAVYKEAMQSERCHRIYLTEIKETYECDAFFPEITSEFQLVKNDDDVPEEIQEENGIQYQYRIYEKLPK

>DHFR_armSub Armigeres subalbatus (mosquito) EU205282 Diptera; Nematocera
MKKFSLIVAVCANGGIGIKGDLPWRLKQELKYFSRMTKKKQDTSKRNAIIMGRKTYFGVPESKRPLPERLNIILTRDPSAHVYPPDVMVCTSMQDALNRLDETPLVDEIENIWIVGGNAVYKEAMQSARCHRIYLTEIKKTFECDAFFPEINSDFQLVKND

>DHFR_danPle Danaus plexippus (monarch) AGBW01009470/AGBW01006850 Lepidoptera
0 MSKVKLNLIAAACENMGIGSNGSLPWRLK 2
1 KEMEYFTTMTTKVKDA SKINAVIMGRRTWDCIPNKNRPLENRLNIVLTRHVDVIKDK 0
1 KEMAYFTTMTTSVSDK KKVNAVIMGRRTWDCIPIKYRPLSNRINIVLTHNVDYVKKQ 0
0 VPKDVVVVSGLDEAVSYIIDRNDVESTWVIGGSSVYQ 0
0 ASMDHPNCGKIYLTEIQKTFDCDTFFPNINRQRFHLVDEDDIPRDIQTEGDISYYFRVYKRL* 0

>DHFR_bomMor Bombyx mori (silkworm) BABH01041437+BABH01041438 Endopterygota; Lepidoptera
0 MSRTQLNLIAAACENMGIGINGTLPWKLK 2
1 KEMAYFTTMTTSVSDK KKVNAVIMGRRTWDCIPIKYRPLSNRINIVLTHNVDYVKKQ 0
0 VPEGVIVVPGLDEAIDHIDSRSDIESTWVIGGSSIYK 0
0 AAMEHPNCGKIYLTEIQKSFDCDTFFPDINKQRFHLVNEEEVPGEKQSEGDINYYYRIYKRL* 0

>DHFR_helVir Heliothis virescens (budworm) AF104106 Lepidoptera
MSQVKLNLIAAACDNMGIGVNGALPWRLKKEMAYFTTMTSKVSEPTKVNAVIMGRRTWDCIPDKYRPLQDRVNIVLTHNVDSVKENVPEGVMVFPGLDEAIKYIEGREDIESTWVIGGSSIYRAAMTHPNCGKIYLTEIQKSFDCDTFFPNIDKQQFHLVDEEQIPGEKQVEGNISYYFRVYKKL

>DHFR_triCas Tribolium castaneum (beetle) AAJJ01000197  Coleoptera exons 2-4 fused
0 MVIKFDLIAAACENMGIGKNNDLPWRLK 2
1 SELAFFSQMTTQTSDESKKNVVLMGRKTWDSIPPKFKPLHQRFNFIMSRAGVNLEGYKDCFSFKSLDEVISKLQDEKFQQLYENVWVIGGSYIYE 0
0 ATMASKYFHRLYLTKVLKTFDCDTFFPKIRDDLIEVRYL 2
1 DPRVPEGVQEENGIQFVYHVYQNPDFNKL* 0

>DHFR_denPon Dendroctonus ponderosae (beetle) GO487443 Coleoptera
 MALKLNLIVAASENWGIGKNGTLPWKLKKEMAFFRTMTSATEDKSKKNVVIMGRRTWESIPAQFKPLPNRINFVLSRSGLNLDQYSNTQSFTNFREALDKLEESAFRQLYENVWIIGGSSLYNETFRSPHFHKLYLTKIHKEFDCDTFLP ALPGNLKEINDPAVTED   IQEENDIEYTFQVFEVIN

>DHFR_apiMel Apis mellifera (bee) HP575683 Hymenoptera
0 MNLHLIAATCEGMGIGIKGTLPWKLK 2
1 SELAFFTYMTTNTKNPNKRNVVLMGRRTWESIPKENRPLKNRINIVLTSQSL 2
1 DYGNDVIVCKNIPHAFEVIEKIKDRIENIWVTGGSSVYK 0
0 EAMESPNFYRLYLTRIKKYFECDTFFPTIPNDFVLTE 2
1 DPNIPQGIQEENDIQFVYEVYKKNRFLIWFQNNIFRIL* 0

>DHFR_bomImp Bombus impatiens (bumblebee) JI114531 Hymenoptera
0 MNLNLIAAACEGMGIGVKGTLPWRLK 2
1 SEMAFFTSMTTNTKDPNKKNVVLMGRRTWECIPKKYKPLKNRINIVLTSQSL 2
1 DYGDDAIVCKSIPHALEVIKKPTLKDQIEGIWVIGGSSVYK 0
0 AAMESPNFHRLYLTRIKKHFVCDTFFPTIPNNFVLID 2
1 DPNIPQGVQEENDIQFVYEVYEKNRFLYLKII

>DHFR_eugCor Euglossa cordata (bee) HP973249 Hymenoptera distal frameshift fixed
MSLNLNIIAAVCEGMGIGVKGSLPWKLKSEMAFFTSMTTQTKDPK KNVVIMGRRTWECIPTKYRPLKDRINVVLTSQNLDYGVNAIICKSMSHALELLKKPPFSDEIESVWVIGGNLVYKAAMESPNFHRLYLTRIKSILTVIHFFPPIPNDFALIQDPNIP

>DHFR_nasVit Nasonia vitripennis (wasp) XM_001605731 Hymenoptera
0 MQVKLKLIAAACENMGIGVNGDLPWRLRK 2
1 EMDFFTKMTSTTKDENKKNVVLMGRRTWESIPKKFKPLSNRINMVLTSQPL 2
1 DLGEDAIVCKNLPEAIEKISQSPLKDKVEQVWVIGGSLVYK 0
0 TSMESPNFYRLYLTRVKKDFKCDAFFPEIPNDYILVK 2
1 DPEVPEGIQEEKDIKFEYEVYEKKQN

>DHFR_copFlo Copidosoma floridanum (wasp) JI834972 Hymenoptera
MQVKLKLIAAACDNMGIGINGDLPWRLRKEMDFFTKMTSTTKDPNKQNVVLMGRKTWESIPKKYKPLANRINVVLSSQPLDLGKDAIACNSLPEAIQQISQDHLRDKVEQVWVIGGSSIYQAAMDSPNFYRLYLTRVKKHFDCDTFFPEIPKDYALVKDPEVPEGIQEEKGINFEYEVYQKA

>DHFR_attCep Atta cephalotes (leafcutter-ant) ADTU01012310  Hymenoptera
0 MPPKLELIAAACENMGIGVNGNLPWRLK 2
1 AEMAYFTRMTTNTRNKNKRNVVLMGRRTWECIPEKYRPLKDRINMVLTSQSL 2
1 NYGDDAIACKSILHALDVISEMQNEVERVWVIGGSSVYK 0
0 SAMESPHFGRLYLTRIKKKFECDTFFPSIPNDLVLIE 2
1 DSTVPQGIQEEKGIEFVYEVYERR* 0

>DHFR_camFlo Camponotus floridanus (carpenter_ant) XP_393902 Hymenoptera 
MPPKLELIAAACENMGIGINGGLPWRLK
TEMAFFTRMTTDTKDKDKKNIVLMGRRTWDCIPEKYKPLENRINMVLTSQSL
NYGDKAIICKSIPHAIEVIAEMQNQVERVWVIGGSRVYK
DVMGSPYFGRLYLTRIKKEFKCDTFFPPIPSGFVLIK
DPTVPQHIQEEKGIKFIYEVYEKK 

>DHFR_harSal Harpegnathos saltator (jumping_ant) XP_393902 Hymenoptera 
MLPKLELIAAACENMGIGIKGDLPWRLK
TEMEYFTRMTTETKDKNKKNVVLMGRRTWDCLPETYRPLRNRINMVLTSQL
MNYGDKAIVCTSIPNALEIITKIQNEVERVWVIGGSKVYK
EAMESPYFGRLYLTRIRKRFECDTFFPPIPNNFVLVQ
DPIVPEDIQEEKGIQFVYEVYEKR

>DHFR_linHum Linepithema humile (argentine_ant) LH21356-RA Hymenoptera
MQLKLELIAAACENMGIGNNGALPWRLK
TEMEYFTRMTIDTKDKNKKNVVLMGRRTWDSIPIKYRPLNDRINMVLTSQSL
DYGDKAIVCKSIPHALEVIAGMQNQIERVWIIGGSSVYK
DAMESPHFGRLYLTRIKKEFECDTFFPPIPNDFVLVT
DPAVPQDVQEEKGIKFVYEVYEKR

>DHFR_pogBar Pogonomyrmex barbatus (harvester_ant) B10193-RA Hymenoptera
MSHKLELIAAACENMGIGINGDLPWRLK
TEMAFFTRMTTETNHTNKKNVVLMGRRTWECIPDKYRPLKDRINMVLTSKSL
DYGNETITCKSISHAFDIITQMQNQIERVWVIGGSSVYK
SAMESPYFGRLYLTRIKKKFECDTFFPPIPNDFVLIE
DPTIPQGIQKENGIEFVYEVYERR 

>DHFR_solInv Solenopsis invicta (fire_ant) AEAQ01011575+AEAQ01011576 Hymenoptera
MPPKLELIAAACENMGIGVNGDLPWRLK
TEMAFFTRMTTDTKQNNKKNVVLMGRRTWECIPKKYRPLKDRINMVLSSQSL
DYKDEAIVCKSIPHALDVISGMQNQVERVWVIGGSSVYK
SAIESPYFGRLYLTRIKKKYECDTFFPLIPNDFVLVE
DPTVPQGIQEENGIEFVYEVYERR

>DHFR_bemTab Bemisia tabaci (whitefly) EZ942456 Hemiptera
MSSLRLSIIVAMTADNGIGFKNSLPWSLPNELRNFAKLTKNCKDKSKQNAVVMGRNTWESIPAQHRPLKDRLNVVLSSTLKDAGEGVLVCPRLKDALTLLEENYQNKIENVWIIGGARVYEDALNLSNCHKLHVTHIQEKPECDVFFPTIPRDFQIITTSEMVEENGFHYNYQVYERRTSPDGEANGIEDSTKS

>DHFR_acyPis Acyrthosiphon pisum (aphid) NM_001162022 Hemiptera one exon
MVYSVIAAVSKNGGIGYKGNLPWKIKKEMEYFNLMTTRVNLKGVQNAVIMGRCTWQSIPDKYRPLKGRINVVISKTLNSVPEGVLLYPKLEEALKSLYLNDRIENLWVIGGSGLYKEAVNDKNCKKLFITKIDQEYLCDTFFPDFDTKKFEETSEANVPKGIQEDNGIKYEFKVFKRL

>DHFR_blaGer Blattella germanica (cockroach) FG125641 Orthopteroidea
MNLKLXLIVAVSENMGIGLNGDLPWRLRKELAHFSRLTKRTIDSNKQNAVIMGRRTWESIPIKNRPLPERLNVVLTKQELDFGKEVLVCNSLNSALEKLCEPRLDKTIESIWVIGGSSVYKEAMSSPHCHRIYLTKIFKTFECDTFLPEIPGNSFKIVRDPDVADEIQEENGLTYEYQVYERILV

>DHFR_pedHum Pediculus humanus (louse) XM_002432121 Phthiraptera
0 MVNNFNLIAAVCSNNGIGYKGNLPWNLR 2
1 KELQYFNRMTKDVKNPEKKNAVIMGRKTWDSLPHNWKPLPGRYNFVLTTQSL 2
1 NLDGAVVCSDLNDLIKKINSPDYSNLIETAWVIGGSKVYESVLKYGLCHRFYLTRIKKEFECDCFFNYDFTKDFKEVS 2
1 DDRVPKGVQKENEIEYVFHVYEKVNESKDSSN* 0

>DHFR_onyArc Onychiurus arcticus (springtail) EW753645 Hexapoda; Collembola
MGPKLYIIAAACENMGIGYKNDLPWKLRNEMAYFNRMTQIVENPGSGSELKNAVIMGRRTWDSIPEKFRPLRNRLNIVLTSRPDFKQPNVLSYPGWYEAVKDLATPEKSLQIDKVWVIGGNSTYKLAVESPDCHRIYLTRVLKHFECDAFFPEFDKSKFRLANDPNVPQDIQEEDGIQYHFEI

>DHFRcalCle Caligus clemensi (sea_louse) BT080999 Crustacea; Maxillopoda; Copepoda
MTSLKCVKINVIAAACKSSRGIGKDNDLPWKLPTDMKYFREKTASTTEIRVGVIMGRRTWESVPPKFRPFKNRFNVVLTSREDFEAKGAVIARTLEDAVKKCSNNVDSIWVIGGSGVYAEALNKYPCRVYLTEIDKEFDCDAFFPPLDEKKYKLASVSPDHQENDITFRFKVYDVIEL

>DHFR_lepSal Lepeophtheirus salmonis (sea_louse) BT121746 Crustacea Copepoda
0 MSKSIPTLHVIAAACRPTNGIGKNNTLPWNLPTDLKYFKTTTSSTPDSKVAVIMGRRTWESIPSKPRPLKNRINVVLTTNSNFVAEGAILA
SSLEEGIKSCMDQADKIWIIGGANLYKEAMQYYKCQIFLTEIDKEFECDTFFPSIDSTKYKEVSVSEDHVENGITFRFKRFERLLQENN

>DHFR_litVan Litopenaeus vannamei (shrimp) FE141783 Crustacea Malacostraca 
MSVRLNIIVAACENHGIGINGELPWKLREEMKYFSRMTKATKSLEKQNAVMMGRKTWESIPAKFRPLPGRLNLVISSRAKDPGPEFNGSTVCASFKEAIDEVQSRTDEVESLWAIGGSSIYE
MALESEHLHRIYLTRILKDFECDTFLPLFDPKKFQIVTD
PAVPAELQQEGDITYKYEIYEKVK

>DHFR_celPug Celuca pugilator (crab) JO488787 Arthropoda; Crustacea
MGPRLNIIVAVAENHGIGKGGELPWRLREEMKHFSRMTKRLDSPATQNVVLMGRKTWESIPLKFRPLPGRLNVVLSSQMKQQQDSSGKVIVCQDFESALKSVSTENPEVETVWVIGGHSVYKMALESEHLHRIYLTKVMKEFECDVFFPSFKMNEFKLVEDPDVPGQV

>DHFR_dapPul Daphnia pulex (water_flea) FE400591 Crustacea Branchiopoda
0 MKLNLIVATASNMGIGFQGTIPWRLK 2
1 KDMALFAKLTKNTNDTNKQNAVVMGRKTWESIPEKNRPLSNRLNIVLSSSQQ 2
1 IENTLTCKSLESAMQLLQTPPYLDQIENIWIIGGASVYK 0
0 EAMEHPSCHRIYVTHILKDFECDVFMPAIDPAKFSLVG 2
1 DSKVPEGTEEENGISFNVKVYEKV* 0

>DHFR_ixoSca Ixodes scapularis (tick) EW930272 Chelicerata Arachnida 3 re-intronated exons
0 MCPGVQESLQYFAIAAMCHNRGIGVLNTLPWRLKKEMAYFSRITSQAAEGKTNAVVMGRNTWDSIPPKYKPLPGR 2
1 VNVVVSRTLEKVPDGHHVARSFSDALSLLQPLVDSGK 0
0 VDKTFVVGGAQLYREALADPRCTRVYLTEIDADFECDVFFPELELGQFRLVQEEGVPQEEQTEGGLTYRFNVYQRISSGSV* 0

>DHFR_ambMac Amblyomma maculatum (tick) JO841362 Chelicerata Arachnida
MCPSHKSVVSCFAIAAMCRNRGIGFKNALPWRLKKEMAFFKRMTSEAAEGKQNAVVMGRNTWESIPPKFRPLNNRINVVVSKTLTEVPEGHHVASSFPAALQLLQTLVDTGKVDKVFLVGGAQLYREALESGHCTRIYLTEVDKDFECDVFFPEFDNSVFSPVEEEGVPQEPQQEDGVTFRFRVYERVQN

>DHFR_varDes Varroa destructor (mite) ADDG01075186 Chelicerata; Arachnida one exon
MAVPPLCAIVAVCKASKGIGNKGSLPWGRSLPNEMKHFARVTTNTTDPDKCNAVVMGRRTWESIPENRRPLPRRINIVISSGLCQESVPEGVKVAKSFDEALEIAGTGCCKGQSRVERIFVIGGTQVYVDAMRHPAMDTVYLTEVLGQFECDTFLNIDTRKFTDIEDPAVTNEEQEENGIRYKYRVLKKIPQ

>DHFR_perSed Peripatopsis sedgwicki (velvetworm) FN242195 Onychophora 
MHRYLNLVVAVCNNLGIGIGGELPWRLRGDMKFFSKLTSETKDSEKTNVVIMGRKTWASIPEKFRPLPKRTNIVISSSLNSVPEGVLIARDFEHAMSLAFGLQQDNIVEQVFVIGGSSVYEQALKSDFPCRIYLTRVMGDFKCDTFFPKLNVDKYALVKVPEIPEEVQEENGISYLFQVLQEKVES

>DHFR_milTar Milnesium tardigradum (tardigrade) EZ761524
--------VAACRNGGIGIRGDLPWTLRGDMRFFTKITSQTKDPQKKNAVIMGRKTWFSIPERFRPLSKRINVVISRTMFGIQPNAIQTQSLEEALDLLSKEPLSSEIETVWIIGGESLYKMAIHSPQLYRVYFTNVMADFECDTFFPTQELQRMKVVSDADVNGDVQMENDVKFQHEIYEP 

>DHFR_triSpi Trichinella spiralis (nematode) ES273161 Nematoda Enoplea
MAQVNIIVAICEKYGIGKKNSLPWHLSKEMQHFKKMTTSVSDPNKINAVIMGRNTWYSIPEKYRPLSGRFNIIISGTMPQLSDEDVIVTWHWREAIEIVEHPISKKAIETFWICGGAKLYNDVIESGLWNRLYVTWIMKEFDCDAFFSFPDKNTIKLVEDDRIPSDIQVEKGIHYKYEVYENCMKMQS

>DHFR_xipInd Xiphinema index (nematode) CV509322 Nematoda Enoplea
MNLIVAACDNMGIGHGNELPWPKLPNESKHFLKLTAGTRDPGKQNAVIMGRKTWEIIPVEHRPLKHRLNVVISRTLSRKDVPPGVQLFEDFEPAMRYLLSPQVSQNIETIWNVGGSEIYALGLDHPSFHRLYFTRVSGTYDCDTYFPQVDWTMLKRVEDPNISTGLHEENGVQYKFETYEHAF

>DHFR_caeEle Caenorhabditis elegans (nematode) NM_059963 Nematoda Chromadorea 4 exons re-intronated
0 MRKMNLIVAMDAEGGIGKNGVLPWRIKKDMQYFASVTKNVSDQSK 0
0 RNAVLMGRKCWESIPVTRRPLAGRLNIVLSRQLPAQKSDDYIVV 2
1 NSLEAAMKLLSEPPFVDSIETIWNIGGAEIYDLALRENLVDEIHLTRIFKN 2
1 FEADVHLKSLDFSKMEKVQNAEVSSENSEIFEENGLKFEFCKWKVVENH* 0

>DHFR_melInc Meloidogyne incognita (nematode) CF803120 Nematoda Chromadorea 3 exons re-intronated
MNIIAAVDENFGIGKNNSLPWRLPKEYKHFINLTTTTKNPNKINAVLMGRKCWESIPEKYRPLKNRLNIVMTKTWVTPEF 0
0 VGENLIFINSLDSLNLILESKPYENLIETIW 2
1 NIGGKQIYSLGIEHQNLNKIVLTKIDKNFDCDVKFPEINWNEFIEEENGEIVEEKGLCWKAITYIKK* 0

>DHFR_ascSuu Ascaris suum (roundworm) JI215038/AEUI01062849/AEUI01051380 Nematoda Chromadorea
0 MVSPKLPINIIVAMDSRGGIGKNGALPWHIPEDLKYFQTMTTKTIDPTKQNAIVMGRKVWESLPAKWRPLKNRLNVVLSNS 0
0 MDDPHDGSYIVTRTFESAIDILNGMSDKIETIWDIGGRRPYEEGLKSSQLRQ 0
0 LYVTFVEGDFDADVFFPDVDFKKFSKQNGDKQSSEHHYEGITYRFETFTALHTDTELS* 0

>DHFR_schMed Schmidtea mediterranea (planaria) DN307116 Platyhelminthes Turbellaria 6 standard exons
0 MKRLNLIVAACENQGIGKNGKLPWNLK 2
1 NEMIYFNNITTSVSDS 1
2 NKQNVVIMGRITWESIPNKFRPMPKRINIVISSTLSR 2
1 DHDNLTGTYLFNNLSEALSFTNDELSDKVEEVFVIGGSSLYK 0
0 AALSSSYPLRLYYTKIFESLDCDTFFPAFEWEQFKSVK 2
1 LDHVPSEVQQENGIRYQFMVLDRC* 0

>DHFR_schMan Schistosoma mansoni (tapeworm) XM_002580511 Platyhelminthes Digenea
0 MRLNVVVAVSENWGIGKGGGLPWKIK 2
1 KDMEFFKTVTTKAHP 1
2 GLKNAVVMGRVTWESIPESFKPLKDRINIVVSSTLSH 2
1 APSFVQVVPSLNAAIDLLYNEEFSSIVDEVFIIGGYRLYK 0
0 EALKQS YPVRIYCTHILSEVDCDTYFPKVDWDKLKK 2
1 VDLPDIPADTFTENGFTFKFCVYDVPSEQFI

>DHFR_taeSol Taenia solium (tapeworm) PEL761437 Platyhelminthes Cestoda
MGLRRLNVIAAVAKNGGIGKENKLPWHIREDMAFFSRITSTAQEGKKNAVVLGRRTWLSFPPKFRPLPNRVNVLVSTQLESVPEGAYLVKSFEDSLRLVESLVDGGEVDEVFIIGGHGLYKEALEQEVYPVRLYYTHIMKDFDCDTFFPSVDWERFTPIQLDTVDSNPKYCGEIEFRFAVYEKGPHRFHDN

>DHFR_aplCal Aplysia californica (sea_hare) AASC02008512 Mollusca Gastropoda one exon
MTSTKLNIVVAVCTNMGIGIEGRLPWRLKQDMAFFKQLTVETQDEQKKNMVIMGKKTWMSIPTKFRPLQDRVNVVLSTQLTEPPSGALLASSLKEAVNLAKSDANVENVFVIGGASVYREAVEGDCPCRIYLTRVDKEFECDTFFPKFDTDVFERIQNPNNVPTGINVEGDLSFTFEVYEKNLFPH

>DHFR_lotGig Lottia gigantea (limpet) FC707856 Mollusca Gastropoda one exon 
MCTKRLNIVVAACKNNGIGVNGSIPWRLKKDMAMFRHITSDTVDESKQNAVIMGRKTWMSIPDKFRPLKNRVNIILSNTLSESPDGTYIVKNFQQALSLVNSDKMKQQIESVHIIGGSSVYKEAMESDYDCRVYLTKVDADFECDTFLPPIEESLFQRVKNPSNIPSDVQEENGIQFTYEIYDKTTDQSDRESPVSR

>DHFR_phyAcu Physella acuta (snail) BW985974 Mollusca Gastropoda 
MPKLNIIVAACNNNGIGIEGRLPWRLKSDMSFFKQITLRTQDAEKKNAVIMGKNTWFSIPSKFRPLVGRINVVLSTQMTDAPEGTQLAKSLNQAVSLLESLNVENIFVIGGSTVYKQAMEGEHPCRIFLTRIAADFLRDTFLPDIPN

>DHFR_pinMax Pinctada maxima (oyster) GT283724 Mollusca Bivalvia
MSKCKLNLVVAACNNRGIGIDGQLPWRLRKDMDFFKKITTETNDPEKRNAVIMGRKTWFSIPEKFRPLSKRINIILSREMKEVPENTYVARSLDEAVQLVSDGGELADRVESIHIIGGSSVYKEALASSHPCRIYFTRVLADFKCDTFLPEFDENKYQKLENMENIPTEIMTEKDINF

>DHFR_mytCal Mytilus californianus (mussel) ES395435 Mollusca Bivalvia
MSKTKSTLNLVVAACNSRGIGINGKLPWRIRKDMDFFKKITMETKNPDKKNVVIMGRKTWFSIPEKFRPLPKRINIILSREMKEAPPGVYIARSFDDAITMVNKDLSDKAESIYVIGGSSIYKAAMDSDYDIRIFLTKILADYDCDTFLPQFDENLYKIIENYEGVPKGKQTENDIDFIFEVYEKVR

>DHFR_dreRos Dreissena rostriformis (quagga_mussel) EY433849 Mollusca Bivalvia
MSMPRLNLVVAICNNNGIGINGSLPWKLRKDMDFFKTITMTTTDPEKQNAVIMGRKTWQSIPENFRPLRNRINIILSRDMNAPNGTHVARSLPDALSLLSSEELADRVEQIHIIGGSSVYKEAMTGPHPCRLYLTRILADFECDAFLPEIDDSSFTKIRNPDNVPTDLQEENGLKFRFEVYDKL

>DHFR_alvPom Alvinella pompejana (bristleworm) GO125274 Annelida Polychaeta 
MPPTLNLIVAMCNNNGIGIQGKLPWKIK
GDMAFFRKMTTETKNPAKMNAVIMGRKTWFSIPEKNRPLNKRMNIILSQTMSSVPDGVHLARSFSEALNLISTKPLADIIEGIFVIGGASVYKEALESSYPCRIYLTRVFADVPCDTFMPTFSGFKQIECAEIDNTVQKENDWQYKFEVYEKV

>DHFR_helRob Helobdella robusta (leech) EY386022 Annelida Clitellata six exons
0 MQPKLQIVVALCVKNRGIGLNNSIPWKLP 2
1 GDMTFFRKLTSETSIL 1
2 GSKNAILMGRKTWDSIPSNLKPLKNRLNVVISRTLE 2
1 cPDInGRLHVSKSFEDAVEFVASDKSIDKIFIIGGSSIYD 0
0 LALASSTCYNVFITEISNDFQCDTFFPKFNQDAYKLIK 2
1 YPGHSTSVQFENGIGYQFTCHHKVSK* 0

Cnidarians, ctenophores, trichoplax, sponge and choanoflagellate reference sequences

>DHFR_nemVec Nematostella vectensis (anemone) ABAV01013911 Cnidaria six exons
MAAVGVRFSCIVAMDLKRGIGKNNDLPWKIK
GDMKFFTEKTSEVKTE
GKQNAVIMGRKVWESIPEKFRPLKGRLNIVLSRTLS
EPPQGAQLCRSFEQALTILSTDPYTKKIENVFVCGGSALYK
DAMAHSACTRIYITYIDQEFDCDVFFPEFDQNTYHLVE
DPDVPSVQHEEKGIKYKFCVYDRCQ

>DHFR_acrMil Acropora millepora (coral) EZ020360 GO002175 Cnidaria Anthozoa Hexacorallia 
VAVADNGGIGKENRLPW HLK
GDMKFFSHLTSTVATE
GKQNAVIMGRNTWESIPKKYRPLPRRLNIVLSRKMS
EPPEGAVLCSSLQDALDRLRSAPYTDTVEKVFVIGGATVYE
EAMQHHQCYRLYITHIYKDFECDVKFPEFDKRIYKETR
DIDVSSEMLEEKGIPYEFKVYQKD

>DHFR_hydMag Hydra magnipapillata (hydra) XM_002157178 Cnidaria Hydrozoa one exon
MVQLRKVHCIAAIDSEWGIGLKGKLPWNLPREYKFFQNITTKVISEGKQNAVIMGKNTWFSIPQQHRPLKNRLNVILSSSFIKEDYPKNVLLESSLEAAILRLSDEFYANTVENIFVIGGSRVYKEAMEKFCDKIYLTKIEQDYSCDVFYPIFDTNKFKEIDDEEVDKNKQIENGVSYTFHVYSSNYTV

>DHFR_mneLei Mnemiopsis leidyi (sea_walnut) AGCP01012173 Ctenophora exons 2-3 fused
0 MKLRFNIIVATDLEGGIGKNNNLPWRCR 2
1 KEFRHFTDTTSKLQNGKQGAAVMGRKTWESIPEKFRPLKNRHNFVISSTLS 2
1 VANPGNVPVYKTISDCMTALDQDKD IGSVWIVGGWGIYK 0
0 EAMESERLHRMYITTVQGKFDCEVFFPSFDERNFNLVI 2
1 DPDIPNETVTEDNGISWSVSVYERK

>DHFR_triAdh Trichoplax adhaerens (placozoa) ABGP01000098 same six exons
0 MQKNFRMIAAMTRDRGIGYKNDLPWKLR 2
1 NEYKYFARLTTSTKDS 1
2 AKKNAVLMGRNTWVSIPEKNRPLRNRINIVISSQLR 2
1 AEEVPEGVHVVASIPEVIELVQSPLLKDIVETVWIVGGAPVYK 0
0 GFMSHPQCDRIYLTKLDANYECDVFFPEMSDDFKEIS 2
1 DPDVSQEVQEENGLKYTFHVYQRN* 0

>DHFR_subDom Suberites domuncula (sponge) GH558547 Porifera Demospongiae
MYIMVAKEIKSGIGNKGKIPWPRINTDMNFLETMTKKTRNKTKQNAAIMGRLTWESIPAKYRPLPERFNIVLSSKKFEDIAPGAHMVCSSLSFAMEKLRAPPLSDQIENVWIFGGAGVYEESMKHPDCDKLYVTEVLQEFPCDAFCPPIDSDIYKLIDHSDDPDVPKGEFVENGVKFKFEVYQRQ

>DHFR_monBre Monosiga brevicollis (choanoflagellate) ABFJ01001321
0 MRVALVVAATKAGGIGHQGQLPWHLK 2
1 ADMQHFRDITTKTEQP 1
2 AAINAVIMGRKTFESIPDKFRPLHGRLNVVLSTQKP 2
1 VSLPLHTPHDTNPPQPLPHPACLAHPQPEVIETAYIIGGASVYN 0
0 EALAKPEYSHYIFMTKVHKDVLCDTVMDMKHLDAYEHDPNFPVq 2
1 * 0

Fungal, microsporidial and amoebozoan reference sequences

Fungi are of special interest because of an existing crystallographic structures in Candida albicans, C. glabrata and Pneumocystis carinii (1AI9, 3CSE, 1CD2). A small sampler of the available sequences is provided below. Microsporidia, obligate intracellular parasites, are today considered a subgroup of 'zygomycete' fungi (for which only DHFR of Rhizopus is available). DHFR is evolving very rapidly within this group -- for example, between two species within the genus Encephalitozoon the percent identity can be as little as 61%.

Encephalitozoon are also odd in having immediately tandem copies of DHFR and TS (thymidylate synthase). These cannot be fused genes because reading frames are out of register (ruling out a linker domain), nor does the intervening DNA contain an intron (which are exceedingly rare in this genus and exhaustively listed). However, a fused DHFR-TS could readily arise from a simple deletion; no such construct has been tested for functionality to date. The basal fungus Rhizopus does not have tandem or fused DHFR-TS.

>DHFR_canAlb Candida albicans (fungi) Dikarya; Ascomycota
MLKPNVAIIVAALKPALGIGYKGKMPWRLRKEIRYFKDVTTRTTKPNTRNAVIMGRKTWESIPQKFRPLPDRLNIILSRSYENEIIDDNIIHASSIESSLNLVSDVERVFIIGGAEIYNELINNSLVSHLLITEIEHPSPESIEMDTFLKFPLESWTKQPKSELQKFVGDTVLEDDIKEGDFTYNYTLWTRK

>DHFR_canGla Candida glabrata (fungi) Dikarya; Ascomycota
MSKVPVVGIVAALLPEMGIGFQGNLPWRLAKEMKYFREVTTLTNDNSKQNVVIMGRKTWESIPQKFRPLPKRINVVVSRSFDGELRKVEDGIYHSNSLRNCLTALQSSLANENKIERIYIIGGGEIYRQSMDLADHWLITKIMPLPETTIPQMDTFLQKQELEQRFYDNSDKLVDFLPSSIQLEGRLTSQEWNGELVKGLPVQEKGYQFYFTLYTKK

>DHFR_pneCar Pneumocystis carinii (fungi) P16184 Dikarya; Ascomycota
MNQQKSLTLIVALTTSYGIGRSNSLPWKLKKEISYFKRVTSFVPTFDSFESMNVVLMGRKTWESIPLQFRPLKGRINVVITRNESLDLGNGIHSAKSLDHALELLYRTYGSESSVQINRIFVIGGAQLYKAAMDHPKLDRIMATIIYKDIHCDVFFPLKFRDKEWSSVWKKEKHSDLESWVGTKVPHGKINEDGFDYEFEMWTRDL

>DHFR_schSti Scheffersomyces stipitis (fungi) Dikarya; Ascomycota
MSEIKPTIAIIVAALKPDLAIGFQGKMPWRLRKEIRYFKDVTTKTSDPSKINAVVMGRKTWDSIPARFRPLPDRINIVLSRSFQNETVDTNVIHANSIANSLSQLQPNVERVFVIGGAEIYNELINDSRVTHLLLTEIENTNEDNQIAVDTFLKFPLYSTDSQWRKQPKSELQKFIGSSITLEDDISEGDLKYNYTLWTRRAG

>DHFR_spaPas Spathaspora passalidarum (fungi) Dikarya; Ascomycota
MTMTEHKPTISIVVAALKPKYGIGYQGKMPWRLRKEIQYFKNVTSKTTKSNGINAVIMGRKTWESIPKKFRPLPDRINIVLSRSFNNETIDDSVIHADSIENGLKLIKGKNVDKVFIIGGGEIYNSVIGSELVDNLLITEIEHTDPETVPMDTFLKFPEDKWVKQPKSELEKFIGEPVDENITEGDYAYNYTLWTRK

>DHFR_lodElo Lodderomyces elongisporus (fungi) Dikarya; Ascomycota
MTSEKPTISIIVAALKPSYGIGNKGKLPWRLRKEMAYFKRVTLRTTHTQNAVIMGRKTWESIPVKFRPLPDRLNIVLSRSFANEEQENGVIHANSIGRCVELIKEKNLLKVFIIGGSEIYHAFLREKGLVDYLLITEIEQDVKNAEGKDKEGKTKEEEVKEVEMDTFLKFDTNEWVMGSQEELKEYTGETEIETNVKEGDFTYNYTIWKRK

>DHFR_debHan Debaryomyces hansenii (fungi) Dikarya; Ascomycota
MTAANGKVTPPIAMIVAALLPELGIGIKGKMPWRLRQEIKYFKNVTTTTKDPNGINAVIMGRKTWESIPTKFRPLPGRLNIVLSRSYVSHGSNDEDIILTNSIENALAKVEEYSKPVEKIFVIGGSELYNKLISHEKVQHLLITEIKSTRPVEVDTWLKFPIYTESSDWTKQTNDDLSKFTGIDNEDVEITEGDFTYKYTYWKKKV

>DHFR_meyGui Meyerozyma guilliermondii (fungi) Dikarya; Ascomycota
MRSVWHRARIYLNHLLPRAFMRQPPIALIVAALLPDLGIGFGGALPWRLKQEIKYFRDVTSNAPDGSINAVIMGRRTWESIPPRFRPLPNRINVVLSRSNPNLEENDVFWGNSFDTALEFLQKRHDINKIFVIGGAEIYNQVINDPRISHLLLTEVSANYDATIPMDRFLHFPREAWTRSPHSQLIQFTGIDATDSTIKEGDFSYNYSLWCKK

>DHFR_milFar Millerozyma farinosa (fungi) Dikarya; Ascomycota
MTSSGHSLGRGGIPIVMIVAALVPEMGIGLKGKMPWRLKNEMAYFRKVTTTTTDNEAINAVIMGRKTWQSIPEKFRPLPKRLNIVLSRSQHENDGDDAVLFCTSLEEALSKAKKYSKPVEKIFIMGGGELYNQAYNSGQVGHLLLTEIRANKEVETDTRIEFPVYGDQSTWIRQPHSALQELVGGEVEEQQLQEGDFTYGFAYFKKKESDE

>DHFR_claLus Clavispora lusitaniae (fungi) Dikarya; Ascomycota
MFSCEPGRSLFISHGCLIISARVSSTWLHHIHMIKSLKPVTSVIVAALSPKYGIGAQGKLPWRLKQEMKYFKDVTSAARAGSINAVVMGRKTWESIPKKFRPLPNRLNIVLSRSFSNEEKDGVLYFNSIDSIMSNLAQSNYWYHDKPIDKIFIIGGAEIYNSVMKGDLVDNLLVTNIRYVGNPEAEPVLDTFLDWDMSLWEQSNVSRIREFSDVEFEEGIIKEGDYEYEYTMWERRK

>DHFR_komPas Komagataella pastoris (fungi) Dikarya; Ascomycota
MSRPFKVNIIVAALIPGHGIGLKGKLPWSLKNEMKYFRLLTTNTISPTSKNVVIMGRNTWESIPSRFRPLQNRLNIVLTRNANISVENGAAVDDSVLVADSLKNGLDIIEKTGNIEEVFLIGGADLYNQCLASGLVDRIFLTEVKSLNGQIETDVSIKIDPSQWIKQESQVLEKYLQDHGVNGFQLDGNKEGELVYDFTLLERR

>DHFR_ogaPar Ogataea parapolymorpha (fungi) Dikarya; Ascomycota
MTKVSLIVAALVPKYGIGYKGQLPWALKEEMRYFRRVTTQTADKNKKNAVVMGRKTWESIPERFRPLKGRVNVVLTRDLSAFSSKYSEEVAKHGNNVKVADSLKSALQTLDMDNIEEVFVIGGAELYNEVLRTTPELVDRLLLTEVSTEKELEMDAFINVGTLWKKDDPQVWKSYLASKGLENEFSQDNREADFQFSYHIYSRC

>DHFR_rhiDel Rhizopus delemar (basal fungus) EIE80550 'zygomycetes'
MTQRKLALIAAATEELGIGRKGDLPWHLPRDLRYFRDVTTKIPKGNSAQNVVIMGRVTWESIPPKYRPLDNRFNIVVSRNPNYDLQTTDPNAILVTSFEEALEKIDATKTNPRVFVVGGAQLYSHAIKRADCTDILLTKIKAKIECDAFFPKIDEHVYRPSTHEELEAYVEESVPEGIQTYKDLEYEFTYYKRMV

>DHFR_encHel Encephalitozoon hellem (microsporidia) CP002713 tandem DHFR-TS
MFALIVAFAKNKGIGKNNRLPWPPLRTDMSWFRTLSQSVPLITSDRIDLSPSSSNAVVMGRKTWDSIPPRFQPLPNRTNIVLSRRPGISTENTLFISAFGDIEHLRIPSSSMVFVIGGHDIYSLAIQSGRAHIIFATEISESLECDVFFPRIDWDAYERRDITREVSGLVGKDLADAFYNTEANAFTENGITFKMFIYIKHGAL

>DHFR_encRom Encephalitozoon romaleae (microsporidia) CP003518 tandem DHFR-TS
 MFALIVALARNKGIGRANRLPWSRPLRTDMSWFRTLSQSIPLITSHHIDLLPSPSNAVVMGRKTWESIPPKFRPLPNRMNIVLSRRPGAKTENAFFIPTFEDLSHLDIPPSSMIFVIGGHDIYNLAIQNGKAQIMFVTEVFESPDCDVFFPHVDWESYEKRDITYEVSRLIDQNLADAFYNTEANAFTENGTTFKMFIYIKPGSL

>DHFR_encCun Encephalitozoon cuniculi (microsporidia) CP003518 tandem DHFR-TS
 MLALVVALASHRGIGNANALPWPRPLAADMAWFRTLSQSIPLISPDRIALAPSASNAVVMGRRTWDSIPSRFRPLANRINVVLSRGPARSTENTFFIQTFEALDSLPLPPSSMTFVIGGRDVYSLALQSGRPHLIFATEVFESPECDVFFPHIDWASYEKRDITRDVSRLIDRTLASAFYSPETATFTENGTSFKMFLYTKPETR

>DHFR_encCun Encephalitozoon intestinalis (microsporidia) NC_014415 tandem DHFR-TS
 MIALIVALAANQGIGRHGKLPWKHPLKTDMAWFRILSQSVPILSPDHISLSPSKSNVVVMGRKTWHSIPSRFRPLQNRINVVLSRSSLPHSQNTFFIPSFAALDHLPLPPSPATFVIGGHDIYALAIQTGRVQAMFVTEVHESPECDVFFPQVDWSSYQKRDITRDVARLVDTTLVDAFYIPEENIFNEGGISFKMFIYTKLHP

>DHFR_nemSpp Nematocida spp (microsporidia) EHY66023
MLPVVLAAYSEGNGVIGREGRLPWPSIKVDFKFMKYLTTKYPSGLIMGRLTYESIGRPLPKRTSIVITSHDKENIQDINYSVLFCKSLESAISICRSLSLQPIIFGGAAVYKEALERYKCKLYLTEIYKIFQGDAFFPLHCIDHRSLVNITPDVLKELSIPENLHRNNVERPLEQSEIYLTQDGRHIPLINTLYENGINYAFLTGYSTPLSEK

>DHFR_harCan Hartmannella cantabrigiensis (slime mold) AAQ02617 Amoebozoa; Tubulinea; Euamoebida pubmed 21112814
MNGFALVVAVTKTMGIGNKGGLPWSSIRGDMNFFKKITSETKDAAKQNAVVMGRKTYVGIPAKFRPLSNRVNIVISTNANLRKEESIPDSVHIVDSFDLAVTKAYNLPNIENVFVIGGSSVYAEAMKHPQCKTIYYTNILTPDFTCDTFFPKIEESVFKVTSRSDLQQEGETSYEFLTYYRI

>DHFR_polPal Polysphondylium pallidum (slime mold) EFA83570 Amoebozoa; Mycetozoa; Dictyosteliida
MVKISFVVAVSLNNVIGKDGGIPWRLKGDLAFFSKTTTGHPIVMGRKTFDSIGRPLPNRHSIVITSSGKLYNRDGIEYQEKPDVLTIVRSTDQAIELVNNLNSDVVYIIGGETIYKETFKYANELLITTVDTVIEGGDTFFNLFPNKATPLIDCNNQSWVIMDKKSFKKDNDNEYDYTISTLKRE

>DHFR_dicDis Dictyostelium discoideum (slime mold) XP_637661 Amoebozoa; Mycetozoa
MKVSIIVAVSKNNVIGTKAGDIPWELPKDLKHFRDTTHGFPCIIGRISLEAFGDLLPNRFNIIVSSKKEFINIREKIAINLPEYETGITEKGAQYVIVKSVKEAVDYAKDILKTDLVYICGGKGIYEDSQQYSHELIITDVDVHIEENDVVIYNPFPSNDERNKWKVVNQIHHPKDHQNNLDFSITYYVKNEEILIDYKVENQN

Plant, water mold, stramenopile, alveolata, euglenozoa, and heterolobosea reference sequences

These species have a fused DHFR-TS proteins that additionally form a dimer (rather than the usual DHFR monomer). This fusion is sometimes taken as a decisive rare genetic event that unites plants with all early diverging eukaryotes. However, a fusions of two genes that act sequentially in metabolism is a very poor choice of synapomorphy because the event could have occurred on multiple occasions as selectively advantageous convergent evolution. For example, much-studied fusions in the fungal tryptophan biosynthetic pathway do not seem phylogenetically informative.

Gene fusions may also be readily reversible -- the long linker region need simply develop a stop codon and initial methionine because the upstream promotor can serve for a polycistronic mRNA. This could be tested experimentally but has not. As noted above, microsporidia have adjacent DHFR-TS separated by a few dozen basepairs.

Finally, a single rare genetic event is unacceptable (the minimal accepted standard is three); rare genetic events have to be comprehensively discovered and checked for phylogenetic consistency.

A careful crystallographic study in Cryptosporidium hominis in comparison to Plasmodium falciparum and Leishmania major, suggested the linker domain in fused DHFR-TS could be used in the phylogenetic classification of protozoa, as the linker seems to control the relative orientation of the two proteins. In particular, a longer leader peptide with very short and conserved linker RN seemed informative.

However as more genomes were sequenced in early diverging protozoa, support for this concept completely evaporates. The set of sequences below show no primary sequence relationships in the linker region that correlate with generally accepted classification. In particular, linker regions are evolving way to fast relative to very old divergences -- linkers are not alignable even within the same genus.

The sequences below are broken into two regions: the DHFR and linker, followed by the earliest conserved residues associated with thymidilate synthase. Fused DHFR have limited relevance to the evolution of non-fused DHFR because interactions of subunits within the fused dimer provide selective constraints that have no counterpart in non-fused DHFR.

>DHFR_araTha Arabidopsis thaliana (thale cress) Q05763 Viridiplantae; Streptophyta bifunctional DHFR-TS 
MTSKPQSTYQVVVAATKEMGIGKDGKLPWNLPTDLKFFKDLTLSTSDSAKKNAVVMGRKTWESIPKKYRPLSGRLNVVLSRSSGFDIANTENVVTCSSIDSALDLLAAPPFSLSIEKVFVIGGGDILREALNKPSCEAIHITEIDTSIDCDTFIPTVDTSAYQPWCSSFPICENGLRFSFTTHVRVKSSSAGEASDESDGSKVLQVDWKKFSSVLPKMIFDR
HEEYLYLNLVKEIISNGNLKDDRTGTGTLSKFGCQMKFNLRRNFPLLTTKRVFWRGVVEELLWFISGSTNAKVLQEKGIRIWDGNASRAYLDGIGLTEREEGDLGPVYGFQWRHFGAKYTDMHADYTGQGFDQLLDVINKIKNNPDDRRIIMSAWNPSDLKLMALPPCHMFAQFYVANGELSCQMYQRSADMGLGVPFNIASYSLLTCILAHVCDLVPGDFIHVIGDAHVYKNHVRPLQEQLENPPKPFPVLKINPEKKDIDSFVADDFELIGYDPHKKIDMKMAV

>DHFR_popTri Populus trichocarpa (cottonwood) XP_002306118 Viridiplantae; Streptophyta bifunctional DHFR-TS 
MSGDSVANLSNGNANTQQSDPRKTYQVVVAATKDMGIGKDGKLPWKLPSDLKFFKDLTLTTMDSGKKNAVIMGRKTWESIPLEHRPLPGRLNVVLTRSGSFDIATAENVVICGSMGSALELLAASPYCLSIEKVFVIGGGQILRESLNAPGCDAIHITEIETDFDCDTFIPAIDTSVFQPWYSSFPKVENDIRHCFTTYVRVRSSTIESHSQNNGENSDGNSENSKFEVKKFSFLPKMVFER
HEEYLYLRMVQDIISDGNLKDDRTGTGTLSKFGCQMRFNLRKNFPLLTTKKVFWRGVVEELLWFISGSTSAKVLQEKGIHIWDGNASREYLDSIGLKDREEGDLGPVYGFQWRHFGARYTDMHADYTGQGFDQLLDVIDKIKNNPNDRRIIISAWNPSDLKLMALPPCHMFAQFYVANGELSCQMYQRSADMGLGVPFNIASYALLTCIIAHVCDLIPGDFVHVIGDAHVYHTHIRPLQEQLRKLPKPFPILKINSEKKDIDTFVAADFKLIGYDPHQKIEMKMAV

>DHFR_selMoe Selaginella moellendorffii (spikemoss) XP_002963028 Viridiplantae; Streptophyta bifunctional DHFR-TS
MVLDGTPPPPAAAATLRAFQVVVAATRDLGIGKEGKLPWSVPSDMAFFKRITSEAADARKKNAVVMGRRTWESIPAKFRPLPGRVNVVLSRAGLELGDKAIVCSSLDSALGILAAPPFDSSVEAVFVIGGGEVFREAMVSPLCTAIHLTEIQTFFECDTFMPALDLTCFRLWYASSPATVNNVRHSFHTYVRNKALDVRNVAGTCSLPIPEFVRER
HEEYQYLNLIKDIIETGNTRSDRTGTGTISKFGCQMRFNLRNSFPLLTTKRVFWRGVVEELLWFISGSTNAKVLEDKGIHIWKGNGSREYLDSIGLSEREENDLGPVYGFQWRHFGARYSTMHDDYTGQGVDQLAEVIDKIKNKPDDRRIVLSSWNPADLKQMALPPCHMFAQFYVSNGELSCQMYQRSCDMGLGVPFNIASYALLTCMIAHVCDLAPGDFVHVLGDAHVYTNHVDPLKEQLENVPKPFPILKIATSNKDIDSFTAKDFQLVDYESHQRIAMKMAL

>DHFR_phyPat Physcomitrella patens (moss) XP_001758934 Viridiplantae; Streptophyta bifunctional DHFR-TS pmid 18079367
MASSGSPAPECDEGEQILSGTHSEKLRGFQIVVAATRERGIGKQGHLPWKLPTDMKFFKTVTSVTTSSSKKNAVIMGRHTWESIPEKFRPLPGRLNVILTRSGIKSTPAGVVVSESLQSALALIATPSYSSHVESVFVIGGGQLVSTVCSEAMASPLCEVIHLTEVEGDVECDTYMPAVDTDIFRIWSASIPIVENGLRISFLTYVRGSLTSSIKQTDNGETTVGLLPKYLLEA
HDEYQYLNLIDDIIKTGAVKGDRTGTGTISKFGCQMRFNLRKSFPLLTTKRVFWRGVVEELLWFISGSTNAKVLHDKGVKIWDGNGSREYLDKQGLTEREEGDLGPVYGFQWRHFGAKYVDMHADYTGQGYDQLKDVINKIKTDPDDRRILLSAWNPADLKLMALPPCHMFAQFYVANGELSCQMYQRSCDMGLGVPFNIASYALLTCILAHVCDLVPGDFVHVLGDAHVYKNHVEPLQEQLQNTPMPFPRLRIKTANRDIDSFVAADFELIGYKAHQKINMIMAV

>DHFR_ostTau Ostreococcus tauri (green alga) XP_003074642 Viridiplantae; Chlorophyta trifunctional dehydrogenase-DHFR-TS smallest free-living eukaryote 
MASYYRALAFALAGARRYGRDGYEAADLEPLREVQNLVGTAFVVTGGSRGIGFAVSEGLIRRGARVHVVCRDEATGASARERLNALGTRTGGSCEVHVSDLSSLAQTRGLVEKLASRGEALHGLVCNAGVMRRERTTSSEGYExxxxxxxDGASRGPQKGPRVTPDASRGLILKSAPAELDGDAVPTSTSTPGRATSSARRLAWGLARGIDALGAFARAGGRSSTGGGGSATRKVRCASARAIGEAKRSRGGMEMRAFQVVVAATVDGGIGKDNGLPWKLTKDMGHFKKLTSETRDANAVNAVVMGRKTWESIPAKFRPLPGRLNVVLSRSGVLAEANDENNNGAETLPEGVLVRKTIDDALSAISSSEKRVEKTFVIGGAQIYEEALKSERCEAVHLTEVEGDFECDVFIPKIDADKFKLYGQSKPITENGARYQFLTYVTKDSESGKFRPIAGELLPAGCDVK
HEEHQYLDMIREIIDQGAVKGDRTGTGTISTFGNQMRFDLRRSFPLLTTKRVFWRGVAEELLWFVAGETNANKLAEKKINIWDGNGSREYLDSIGLTEREVGDLGPVYGFQWRHFGAEYTNMHADYTGKGVDQLAEVINKIKNNPTDRRILLTAWNPAALKEMALPPCHMFCQFYVANGELSCQMYQRSCDMGLGVPFNIASYSLLTCMIAQVCDLKPGDFVHVCGDTHVYSNHVEPLKKQLTCEPRPFPILKINPEKKDIDSFTFDDFEIIGYDPHPKIEMKMAV

>DHFR_micPus Micromonas pusilla (green alga) XP_002502546 Viridiplantae; Chlorophyta bifunctional DHFR-TS
MASEASTVRPFQVVVAATKGEMGIGYQGGLPWNLPKDMAYFKAVTSQVNEPGTKNAVVMGRKTWESIPAKFRPLPGRVNVVLSRGHTLVEAGSENSAPVNGGGKPEVLPEGVFLRPSLEAALELLSAPEHEAGVEKVFVIGGAQVYAEAMASPHCQALHLTEVTPPADEPEKYKCDAFLPQIDPAKFKLYASAKPLREKDGATIQFLTYFGVDPGTGKFRSPGSKVLPAGAVAKGVR
HEEMQYLDLIKEIMEEGNVKGDRTGTGTISKFGCQMRFDLRRSFPLLTTKRVFWRGVAEELIWFVKGCTSAKELQDKDIHIWDGNGSREYLDSVGLGHREEGDLGPVYGFQWRHFGAEYKDMHADYTGQGVDQLAEVIDKIKNNPNDRRILLTAWNPAALKEMALPPCHMFCQFYVANGELSCQMYQRSCDMGLGVPFNIASYSLLTCMVAQVCGLKPGDFVHCCGDTHVYSNHVEPLYKQLENEPRPFPTLKINPEKKDIDSFEFSDFEIVDYDPHPKIAMQMAV

>DHFR_chlVar Chlorella variabilis (green alga) EFN56442 Viridiplantae; Chlorophyta bifunctional DHFR-TS
MTPEQARRTIQIVVAATKQWGIGKGGSLPWSLPGDMKYFRELTSRTADPAKQNVVIMGRKTWESIPAKFRPLAGRINVVLTRGAVAGDENASAPGNATAALAEASKAEGVHISSSLDSALEMLSGPEFDSRVESVFVIGGGQVYKECMESPLLSAIHLTLVEGEAAEAANCDTFMPPVDESRFRLWSASAPRSVGGTRYSFLCYTRAGQEGLPALPPTVASR
HEEHQYLEMIEDVMANGVFRGDRTGTGTYSKFGATMRFNLRHSFPLLTTKRVFWRGVAEELLWFVSGCTNARVLMDKGIHIWDGNGSREYLDSIGLSHREEMDLGPVYGFQWRHFGAEYTDMHADYGGKGVDQLAELIHKIKTNPNDRRLVLSAWNPAALTDMALPPCHMFCQFYVADGELSCQMYQRSCDLGLGVPFNIASYSLLTCMVAQVCGLRPGDFVHVLGDAHVYANHVGPLRDQLKNAPRHFPRLLINPAKTDIDSFVFDDFQLEGYQPHKAIKMQMAV

>DHFR_volCar Volvox carteri (green alga) XP_002955893 Viridiplantae; Chlorophyta bifunctional DHFR-TS
MTSHKGFQLVVAATPSLGIGKNGTLPGWKLPGDMAYFRELTSRTREPSCQNVAIMGRKTWESIPNKFRPLKGRINIIARRNGAQFGPDVLGCSSLGAALELLEDEKLKRDVDFVFVIGGAQVYAEALRHPSCTAVHLTKVEKEFDCDAHLPPLDPSVFGVWSASEPFQENDTRYSFVCYTRRGLSPPPELPPGMASR
HDEIQYLELVRELITSGTFRPDRTGTGTYSRFGRTARYNLRHTFPLLTSKRVFWKGVAEELLWFIRGSTNANLLRDKNIHIWDGNSTREYLDSRGLHHREVGDLGPVYGFQWRHFGAAYKDMHTDYTGQGVDQLKAIIEQIRKDPNDRRMIMSAWNPAALNEMALPPCHMFCQFYVADGELSCLMYQRSCDVGLGVPFNIASYALLTRLVAQVTGLRAGDFIHVMGDTHVYANHVEPLKEQLKNTPRHFPILNINPAKTDIDSFVFEDFELVDYQPHQTIKMKMAV

>DHFR_chlRei Chlamydomonas reinhardtii (green alga) XP_001700389 Viridiplantae; Chlorophyta bifunctional DHFR-TS
MDTSRKGFQLVIAATPSLGIGKNGKLPDWQLPGDMAYFKELTSRTRDGAHTNAVIMGRKTWESIPAKFRPLKGRLNIVLSRSFADDADLLESALSLLEHKELKGGVEGVFIIGGGQVYAEALAHPNCTAVHLTQVEKEYECDTFLPALDPAVWGVWSSSEPVTENNTRYSFVCYTRRGTEQAPELPPCMASR
HDEQQYLELVRELITSGVFRPDRTGTGTYSRFGRTSRYNLRHTFPLLTSKRVFWKGVAEELLWFISGATNANLLRDKNIHIWDGNSSREFLDGRGLGHREVGDLGPVYGFQWRHFGAEYKDMHTDYTGKGVDQLRYIIDTLKKDPNDRRMVMSAWNPAALAEMALPPCHMFCQFYVADGELSCLMYQRSCDVGLGVPFNIASYALLTRLVAQVTGLRAGELVHVMGDTHVYANHVEPLREQLKNTPRHFPTLRINPDKKDIDSFVFEDFELVDYNPHKTIKMQMAV

>DHFR_phyInf Phytophthora infestans (water mold) XP_002904741 stramenopiles; Oomycetes bifunctional DHFR-TS VLCKLSWSPSLVGVVYLVKADEGA*
MTEQSIRVVVAALETTGGIGLRQHIPWRLPSDMKHFRALTTASPDSSVQHAVIMGRKTWESLPAKVRPMPKRYNVVLTRDSSYRNKQEIPDTVGVAASFREALKLVQEQGDKVDQVFVIGGGAVYAEALSYPSCTKVHLTRVKGQFECDAFFPLEQLEQNFKVAEESEVKEENGVQFHFVEWERKTGEVKAVESALLVDNTTP
HEEMQYLDLIRKILTQGAKREDRTGTGTLSVFGAQMRFSLRNNVFPLLTTKRVFWRGVAEELLWFISGDTSAHTLQQKDIHIWDGNGSREYLDSRGLQHREVGDLGPVYGFQWRHFGAKYTDMHADYTGKGVDQLAEVIHKLRTNPTDRRIVLSAWNPSDLNEMALPPCHMFCQFYVANGELSCQMYQRSADMGLGVPFNIASYALLTRLVAQVTGLKPGEFIHVIGDAHVYLNHVEPLQKQLTRTPCPFPTLHINPEKTSSIDEFTFEDFEWVSDYAGSAIkmkmai 

>DHFR_albLai Albugo laibachii (white rust) CCA17727 stramenopiles; Oomycetes bifunctional DHFR-TS
MVRTISIVVAATEDCWGIGWRQSIPWKLAKDLKHFRDLTTRTFSFNGCNSQNMRNAVIMGRKTWESLPPSYQPLPNRYNHILTRTTNYRIDHSVPHDVGISSSLADALEEIERQEDIGRVFIIGGQKVYEAALECTSCDRIYLTTVKATYQCDTFFPSNLKDRGFQLVSESDEMEENEIRFHFRELSRPIVTITKHSGLYTNEKP
HEEYQYLNLIRNIVANGVEKQDRTGVGTKSIFGVQMRFNLRNDMFPLLTTKKVFWRGVAEELLWFISGNTNAKTLQDRKVRIWDGNGSSAYLKSIGLHHREEGDLGPIYGFQWRHFGAKYTDMHADYTGQGFDQLADVILKLKTDPNGRRIILSAWNPNDLNEMALPPCHLMCQFYVAKGQLSCQMYQRSADMGLGVPFNIASYALLTKMIAHVTYLTPGEFVHVIGDAHIYLNHLEPLKLQLERTPRPFPTLLIRPRNDLRTIDDFEFTDFELQNYEPYMAIKMAMAV

>DHFR_blaHom Blastocystis hominis (protozoan) CBK20227 stramenopiles; Blastocystis bifunctional DHFR-TS WSVCLNQWFI*
MSIIRFSIVAAMTTKRGIGLNGGLPWRIKQDMKFFVDLTTTTTDSEKQNAVIIGKNTYFSFPEKFRPLKNRINFIISHDKELREKHNIPDSVYICSSVDEALILMQEEELKRKIENIFVIGGGQIYKKAINLPECEKLYLTEVDADISCDTFFPEIPLAYKKTKESETFEENNFKFRFAEYTRLPMTFLPPYRTSLQE
HEELQYLNLIRYILDHGALRTDRTNTGTRSVFGTQMRFLLANGVIPLLTTKKVFWRGIVEELLWFIRGCTDGRVLKEKNIHIWDGNGTRSFLDSVGLTQNEEDDLGPIYGFQWRHFGAAYEGRDKDYSNQGVDQLKEVIHLLKTDPYNRRIILSAWNPADLRKMALPPCHVMSQFYVQNGYLSCQMYQRSADMGLGVPFNIASYSLLTILLAHITGLKPGEFVHSIGDTHVYVNHEEALLQQIEREPRPFPTLRITKQSENIEEYTFADFELVGYNPHETLKMKMAI

>DHFR_ectSil Ectocarpus siliculosus (brown alga) CBN76613 stramenopiles; Phaeophyceae bifunctional DHFR-TS
MASSVAVASPSTSRSFVVIVAATAGSLGIGKNGALPWRLAADMAYFKRCTSTPTTSSGTTTATTTTDKINAVIMGRKTWQSIPERFRPLAGRRNVVLSRNPAARDTLRLPKDVLLAGSLTEALDLLSPAAASEVDKVFVIGGGSVYAEAVASELCEKVLLTSVTPPADGRFDDCDTHFPALDPDTFKLVKKGEAQEEKGVELSFDEYESVNHSEAAAAMAAAEGDKENASAASVVGSKRKSSPDGDQVAATGVETPAAAAAAGDGGATADAAAAAGSASTAPVPQEEAEGGPGN
EEEMQYLNLVEDILDNGVRRGDRTGTGTLSKFGVQMRFSLREERFPLLTTKRVFWRGVAEELLWFVAGCTNANVLKERGIHIWDGNGSREFLDGLGLTEREEGDLGPVYGFQWRHFGAEYTDMHADYTGKGVDQLAECIQKIKTNPEDRRIVLSAWNPADLGKMALPPCHMFCQFYVAEGELSCQMYQRSADMGLGVPFNIASYALLTRLVAQACGLKAGDFVHTIGDAHVYVNHVDALREQLKRKPRPFPTLSINTDETDIDSFEYKHLEVKGYSPYKTIKMKMAV

>DHFR_aurAno Aureococcus anophagefferens (microalga) EGB04584 stramenopiles; Pelagophyceae bifunctional DHFR-TS
MRPALTKMAPTNDLPQLTCIVAAVAGSYGIGKDGTLPWKLAGDMKYFKKVTSAAPEGKTNACVMGRKTWLSIPPKFRPLGGRKNVVLSRNPNAREELGLPPDVFVCPSLDAAVDALAGEADLGEVFVIGGGQVYASALADARCTKVLLTSIDDAAGAFADCDAVFPDPAKAGFASAASGEALVENGVAYRFETLTRPAKKPPLRESNGPPAPPPANEKPAAPN
HEEKQYLDLIAEILDRGVRRGDRTGTGTISIFGARMSFSLRNDRFPLLTTKRVFWRGVAEELVWFVSGSTNGKLLADKGIHIWDGNGSREFLDRRGLGHREEMDLGPVYGFQWRHFGAKYVDMHKDYAGEGVDQLAACVETIKQDPNSRRIVLSAWNPADLDEMALPPCHMFCQFYVANGELSCQMYQRSADMGLGVPFNIASYALLTRMVAQATGLKPGDFVHVIGDAHVYSNHVDALKVQLQREPRPFPTLTINPAKTGIDDFEFADFTVEGYKPYPKIDMKMAV

>DHFR_phaTri Phaeodactylum tricornutum (diatom) XP_002177415 stramenopiles; Bacillariophyta bifunctional DHFR-TS frag
VVAAAASHRGIGYQGSLPWRLPGDLRHFQAVTTQPPETGGTNAVIMGRKTWDSIPDRFRPLPGRINVVLTRHPERTLWPDGVLTATSLADAAALLAQPTRAVGHVFVIGGGQVYEQALREKFVNRVVYTEVANLPADSKFDAFFPPLPDAEWQAQPWQTSTTATNKENGNAPETGAWHTDGPSGITYRFLEYTRKVASVTVQPPTDGN
AEERQYLDLCRHILDHGVRRGDRTGTGTLSQFGTQMRFSLRDGTLPLLTTKRVFWRGVAEELLWFVAGNTNANDLAAKDIRIWDGNGSRDFLDSRGLGHREVGDLGPVYGFQWRHFGATYGTMHDDYTGKGVDQLAECIDKIKNNPEDRRIILSAWNPADLELMALPPCHMFCQFYVRTASLSCQMYQRSADMGLGVPFNIASYALLTHMIAHVTGRKPGDFVHTIGDAHVYLNHVDALQTQLERTPRAFPKLQINPDKQNIDDFVFADFTVVGYQPHKTIKMEMAV

>DHFR_thaPse Thalassiosira pseudonana (diatom) XP_002295808 stramenopiles; Bacillariophyta bifunctional DHFR-TS
VVAAAAGSRGIGHQGKLPWRLPGDMNHFKKVTTTPPSPGLTNAVIMGRKTWDSIPSKFRPLDGRVNVILSRKSAAGVEGVPMSTDDNNNNNNAEDVLVASSLEEAMQKLDGRANHGSTFIIGGGEIYSQGIKSGLVKRVIYTNVKGLAEDAKLDAFFPELTDSEWECVPFTPKDNAENEETENNHVSKKAKVAEEHEDAKSGLKYEFLEYIRRDNVDIPEGADVNPEEMQYLEICRDIIENGVRRGDRTGTGTLSKFGVQMRYSLRDNTLPLLTTKRTFWRGVAEELLWFVKGSTNANELADKNIHIWDGNGSREFLDSRGLQHREEGDLGPVYGFQWRHFGAEYKDMHADYTGQGVDQLADCIDKIINNPEDRRIVMSAWNPKDLDVMALPPCHMFCQFYVDTDRNEVSCQMYQRSADMGLGVPFNIASYALLTHMIAKVTGRKAGDFVHTIGDAHVYLNHVDALKEQLERKPRAFPKLKMKDGKDFNDIDGFEFDDFEVVGYKPHKTIKMKMAV

>DHFR_cryHom Cryptosporidium hominis (protozoan) 1QZF Alveolata; Apicomplexa bifunctional DHFR-TS 11 aa linker
MSEKNVSIVVAASVLSSGIGINGQLPWSISEDLKFFSKITNNKCDSNKKNALIMGRKTWDSIGRRPLKNRIIVVISSSLPQDEADPNVVVFRNLEDSIENLMNDDSIENIFVCGGESIYRDALKDNFVDRIYLTRVALEDIEFDTYFPEIPETFLPVYMSQTFCTKNISYDFMIFEKQEKKTLQNCDPARGQLKSIDDTVDLLGEIFGIRKMGNRHKFPKEEIYNTPSIRFGRE
HYEFQYLDLLSRVLENGAYRENRTGISTYSIFGQMMRFDMRESFPLLTTKKVAIRSIFEELIWFIKGDTNGNHLIEKKVYIWSGNGSKEYLERIGLGHREENDLGPIYGFQWRHYNGEYKTMHDDYTGVGVDQLAKLIETLKNNPKDRRHILTAWNPSALSQMALPPCHVLSQYYVTNDNCLSCNLYQRSCDLGLGSPFNIASYAILTMMLAQVCGYEPGELAIFIGDAHIYENHLTQLKEQLSRTPRPFPQLKFKRKVENIEDFKWEDIELIGYYPYPTIKMDMAV

>DHFR_toxGon Toxoplasma gondii (protozoan) Q07422 Alveolata; Apicomplexa bifunctional DHFR-TS
MQKPVCLVVAMTPKRGIGINNGLPWPHLTTDFKHFSRVTKTTPEEASRLNGWLPRKFAKTGDSGLPSPSVGKRFNAVVMGRKTWESMPRKFRPLVDRLNIVVSSSLKEEDIAAEKPQAEGQQRVRVCASLPAALSLLEEEYKDSVDQIFVVGGAGLYEAALSLGVASHLYITRVAREFPCDVFFPAFPGDDILSNKSTAAQAAAPAESVFVPFCPELGREKDNEATYRPIFISKTFSDNGVPYDFVVLEKRRKTDDAATAEPSNAMSSLTSTRETTPVHGLQAPSSAAAIAPVLAWMDEEDRKKREQKELIRAVPHVHFRG
HEEFQYLDLIADIINNGRTMDDRTGVGVISKFGCTMRYSLDQAFPLLTTKRVFWKGVLEELLWFIRGDTNANHLSEKGVKIWDKNVTREFLDSRNLPHREVGDIGPGYGFQWRHFGAAYKDMHTDYTGQGVDQLKNVIQMLRTNPTDRRMLMTAWNPAALDEMALPPCHLLCQFYVNDQKELSCIMYQRSCDVGLGVPFNIASYSLLTLMVAHVCNLKPKEFIHFMGNTHVYTNHVEALKEQLRREPRPFPIVNILNKERIKEIDDFTAEDFEVVGYVPHGRIQMEMAV

>DHFR_babBov Babesia bovis (protozoan) AAP57962 Alveolata; Apicomplexa bifunctional DHFR-TS 
MSTVYEGCGGLTIYVAIALNRVIGHQNQIPWPHIIHDFRFLRNGTTYIPPEVLSKNPNIQNVVIFGRKTYESIPKASLPLKNRINVIISRTVKEVPGCLVYEDLSTAIRDLRANVPHNKIFILGGSYLYKEVLDNGLCDKIYLTRINKEYPGDTFFPDIPDTFEITAISPTFSTDFVSYDFVIYERKDCKTVFPDPPFDQLLLKGVDIIVPKPKYVACPGVRIRN
HEEFQYLDILADVLSHGVLKPNRTGTDAYSKFGYQMRFDLSRSFPLLTTKKVALRSIIEELLWFIKGSTNGNDLLAKNVRIWELNGRREFLDKNGFTEREEHDLGPIYGFQWRHFGAEYLDMHADYTGKGIDQLAEIINRIKTNPNDRRMIVCSWNVSDLKKMALPPCHCFFQFYVSDNKLSCMMHQRSCDLGLGVPFNIASYSILTAMVAQVCGLGLGEFVHNLADAHIYVDHVDAVTTQIARIPYPFPRLRLNPDIKNIEDFTIDDIVVEDYVSHPAIPMAMSA

>DHFR_plaFal Plasmodium falciparum (malaria) AAA29585 Alveolata; Apicomplexa bifunctional DHFR-TS
MMEQVCDVFDIYAICACCKVESKNEGKKNEVFNNYTFRGLGNKGVLPWKCNSLDMKYFCAVTTYVNESKYEKLKYKRCKYLNKETVDNVNDMPNSKKLQNVVVMGRTNWESIPKKFKPLSNRINVILSRTLKKEDFDEDVYIINKVEDLIVLLGKLNYYKCFIIGGSVVYQEFLEKKLIKKIYFTRINSTYECDVFFPEINENEYQIISVSDVYTSNNTTLDFIIYKKTNNKMLNEQNCIKGEEKNNDMPLKNDDKDTCHMKKLTEFYKNVDKYKINYENDDDDEEEDDFVYFNFNKEKEEKNKNSIHPNDFQIYNSLKYKY
HPEYQYLNIIYDIMMNGNKQSDRTGVGVLSKFGYIMKFDLSQYFPLLTTKKLFLRGIIEELLWFIRGETNGNTLLNKNVRIWEANGTREFLDNRKLFHREVNDLGPIYGFQWRHFGAEYTNMYDNYENKGVDQLKNIINLIKNDPTSRRILLCAWNVKDLDQMALPPCHILCQFYVFDGKLSCIMYQRSCDLGLGVPFNIASYSIFTHMIAQVCNLQPAQFIHVLGNAHVYNNHIDSLKIQLNRIPYPFPTLKLNPDIKNIEDFTISDFTIQNYVHHEKISMDMAA

>DHFR_thePar Theileria parva (parasitic protozoan) EAN31856 Alveolata; Apicomplexa bifunctional DHFR-TS
MAEDYSGLPFVKLLVAITPENGIGISNGLPWPHIKRDFLFMFRATTYVDPEFKEKHPGAQNVIIIGRKTYDSLPEGTFPLKNRINVVISRNLTEVPGGLVFPSMPAAIRHVTEHLNYYKIFIMGIFHLNIKHLGGSYVYRDALDARIVDKMYVTRVNKNYKCEVFFPKIPDDFVITGISKTFSSEDTTYDFVTYERKDLIVDDRVKKTFEEMLLTGEKFATKDQLVKACPSLKVRY
SQELQYLEILADILNTGYHKKNRTGVDCIAKFGYQMRFDLSESFPLLTTKKVFLRGIIEELIWFIKGSTNGNELLKKNVRIWELNGARSFLDNLGFFDREEHDLGPVYGFQWRHFGAKYVDMHTDYSNQGIDQLSLVIDRIKNNPDNRRLIVCSWNVADLSKMALPPCHLLFQFFVADGTLSCMMHQRSCDMGLGVPFNIASYSLLTYMIAQICGLKPGEFVHNLGDAHIYNNHIDAIKEQISRVPYPFPTLRLNKDITRIEDFTIDDIKVENYRSHPVIKMEMAA

>DHFR_perMar Perkinsus marinus (protozoan) XP_002768310 Alveolata bifunctional DHFR-TS
MALPQLSVIVAHTCKWGIGKDGQLPWKSLPEDMKRFKKITTGGHCNDNVKNVCIMGRKTWESIPERFRPLRDRINVVISSTTSPTDYPPGVRICPSLLDSLASLQQGEVEIGEIFVIGGASLITEAMQLPQCRTLYTTRVGTDPWECDVFMEKIDEKIWEPIAVSRTFRHSDIPYDFVDYKRREGSDKTELSLAERCPRGKFQ
HGEYEYLRLIEEIIENGEDADDRTGVGTKTIFSCHMRFSLRDGVFPLLTTKRVFWRGVVEELLWFIRGDTNAKHLSDQGVKIWDLNGTREFLDGRGLTHREEGDLGPVYGFQWRHFGADYVDMHADYSKSGGVDQLAEVIRQLKENPMDRRIIMTAWNPTALPLMALPPCHMMCQFYVNSKQELSCAMYQRSADMGLGVPFNIASYALLTCMLAQVCGLKPGEFCHHLGNTHVYNNHIEPLRQQLKRTPRPFPLLKINPNVKTIDDIKSSDFELIGYNPWGKIAMEMAV

>DHFR_parTet Paramecium tetraurelia (ciliate) XP_001453677 Alveolata; Ciliophora bifunctional DHFR-TS
MTTNKTFSMVLAMTLNGGIGYQNRLPWKLKEDLQRFKKITTGGIVIMGRKTFESMNSKPLPNRVNVVISKNMKSSNEVQVFPRIEDALQFYNTSHQKLYLIGGKRIFEEGLATDKCSDVHLTRIGVETKCDVYLNKNIFSTFQVNKTSQTKSENGINYDYQHLINKNSHEQSYIDEE
HQENQYLDMITKIMKEGVSKDDRTGVGTMSIFGQTMRFNLAQSFPLLTTKKVFFRGVVEELLWFLRGNTNGKLLLDKGVKIWEGNGTREYLDTIGLQHRQEHDLGPVYGFQWRHFGAKYKDCQTDYSNQGVDQVKEIIQLLKNNPDSRRIILSAWNPIDLKQMALPPCHVMSQFFVANGKLSCMMYQRSCDFGLGIPFNIASYALLTYMLAKECNLNLGEFVHVLGDTHIYSNHVEALKKQIERVPYPFPLLKIKGNKSLFDYTYEDFELVGYNAHDKIEMKMAV

>DHFR_tetThe Tetrahymena thermophila (ciliated protozoan) EAR85731 Alveolata; Ciliophora bifunctional DHFR-TS
MKTRHFDIVLAQTLKKQGIGYKNSLPWRLPNELKNFKKITTETKNKGLQNAVIMGKNTWEALPKKQQPLKDRLNIVISTTMQEGQIADHSYACKSLDSALNFLEQQNQIQDALVIGGAKLCQQALSDQRLRQIHLTRVGVEVECDVFMQKDYLKNFDMIEVSETQSENNLNYDFTRYFNKNYKGQVDPSLFKKMYKP
HQEYQYLELIDEIIKNGHVKTDRTGTGTISQFGKLMRFDLSKSFPLLTTKNVFWRGVVEELIWFIKGSTNSKILSEKGVKIWDGNGSREFLDQLGFKNREEGDLGPVYGFQWRHFGAEYKDMHTNYKGKGVDQLQDLINTIKKNPDSRRMIMNAWNVKDLPLMALPPCHVMSQFYVNDNKLSCMMYQRSCDMGLGIPFNIASYALLTHMIAQVTNMQVGEFIHVLGDAHVYSNHVDQLKIQLERAPYPFPLLKINNNKQYNSIEDFTLEDFELIGYNYHPKIQMKMAV

>DHFR_tryCru Trypanosoma cruzi (trypanosome) AAX40325 Euglenozoa bifunctional DHFR-TS short RN linker
ETVAEGTRLALRAFSLVVAVDEHGGIGDGRSIPWNVPEDMKFFRDLTTKLRGKNVKPSPAKRNAVVMGRKTWDSIPPKFRPLPGRLNVVLSSTLTTQHLLDGLPDEEKRNLHADSIVAVNGGLEQALRLLASPNYTPSIETVYCIGGGSVYAEALRPPCVHLLQAIYRTTIRASESSCSVFFRVPESGTEAAAGIEWQRETISEERTSANGNETKYYFEKLIPRN
REEEQYLSLVDRIIREGNVKHDRTGVGTLSIFGAQMRFSLRNNRLPLLTTKRVFWRGVCEELLWFLRGETYAKKLSDKGVHIWDDNGSRAFLDSRGLTEYEEMDLGPVYGFQWRHFGAAYTHHDANYDGQGVDQIKAIVETLKTNPDDRRMLFTAWNPSALPRMALPPCHLLAQFYVSNGELSCMLYQRSCDMGLGVPFNIASYALLTILIAKATGLRPGELVHTLGDAHVYSNHVEPCNEQLKRVPRAFPYLVFRREREFLEDYEEG

>DHFR_leiTro Leishmania tropica (trypanosome) AAM88659 Euglenozoa bifunctional DHFR-TS short RN linker
MSRAAARFKILMPETKADFAFPSLRAFSIVVALDKQHGIGDGESIPWRVPEDMAFFKDQTTLLRNKKPPTEKKRNAVVMGRKTWESVPVKFRPLKGRLNIVLSSKATVEELLAPLPEEKRAAAEQDVVVVNGGLAEALRLLARPPYCSSIETAYCVGGAQVYADAMLSPCVEKLQEVYLTRIYTTAPACTRFFPFPPENAATAWDLASSQGRRKSEAEGLEFEICKYVPRN
HEERQYLELIDRIMKTGIVKEDRTGVGTISLFGAQMRFSLRGNRLPLLTTKRVFWRGVCEELLWFLRGETNAXLLADKDIHIWDGNGSREFLDSRGLTENKEMDLGPVYGFQWRHFGADYKGFEANYDGEGVDQIRFIVETIKANPNDRRLLVTAWNPCALQKMALPPCHLLAQFYVNTDTSELSCMLYQRSCDMGLGVPFNIASYALLTILIAKATGLRPGELVHTLGDAHVYRNHVDALKAQLERVPHAFPTLIFKAERQFLEDYELTDMEVIDYVPHPPIKMEMAL

>DHFR_criFas Crithidia fasciculata (trypanosomid parasite) AAA30318 Euglenozoa bifunctional DHFR-TS 1989
MSRAAAKFKIPMPVTKADFAFPSLRAFSIVVAADQQHGIGDGETIPWTVPETLAFFKDQTTLLRNKKPPTEKKRNAVVMGRKTWEVPLKFRPLKGRLNVVLSCRRPVDDLLAQLPEEKRAAAAADVVINGGLKEALHLLARPPYCSSIETAYCIGGARVYTEAMQSPCVEKLKEVYLTRVHTAPTCNRFYEFVRRRRARAAVDLESTSGVKVSDAAAHLSYEIMKYVPHN
AEERQYLELIDRIMKTGLVKEDRTGVGTISLFGAQMFSLRDNQLPLLTTKRVFWRGVCEELIWFLRGETNAHVLADKDIHIWDGNGSREFLDSRGLTENKEMDLGPVYGFQWRHFGADYKGFDANYDEGVDQIKTIVETLKTNDRRLLVTAWNPCALHKMAVRPCHLLGQFYVNTQTKELSCMLYQRCCDMGLGVPFNIASYALLTILIAKATGLRPGELVHTLGTAHVYSNHVEALKEQLQRVPVAFPVLVFKKEREFLEDYESTDMEVVDYVPYPPIKMEMAV

>DHFR_naeGru Naegleria gruberi (amoeboflagellate) XP_002668828 Heterolobosea; Schizopyrenida bifunctional DHFR-TS DIIKEl
MMQSTSNLPKASSIPPMEAVVAVLLNSNGIGLNGNLPWLQDGLTEDMKHFVSVTCSSEPLKFISPSMYKQSSSSSPITSDNTNDNTTTINDQLMNAVIMGRKTWESIPSKFKPLSKNRHAIILSSKVNDDDQLMMERTTFVKNVDELFELLQNQKFNRIFIAGGTSIYKLLLPYTAVIHYTNIIEQKGVNIQVDTRFPIRLLPSSDYRNVPINSNQFIAVDKSDKILPKNNERISHYEFITFERR
HEEFQYLDIIKELILSDINVREERTGVGTVSKFGYQMKYSLRNNVFPLLTTKRVFWKGVAKELFWFISGSTNAKILSEQDIHIWDGNGSREYLDSLGFENREEMDLGPVYGFQWRHWGSSYENMHTDYTNKGVDQLKKCIETLKNNPTNRRIILSAWNVSDLPNMALPPCHLMCQFWVDTDTNELSCQMYQRSCDMGLGVPFNIASYALLTRLMAQVCNLKPGDFIHTLGDTHVYKNHIEPLKKQIERDPKHFPLLLLDPSITNIDDFKFEHLTLQNYEPHATIKMEMAV

Bacterial reference sequences

Bacterial DHFR, especially that of E. coli, have been intensively studied for decades. A small sampler of the available sequences is provided below.

>DHFR_escCol Escherichia coli (bacteria) M84522 gammaproteobacteria
MNSESVRIYLVAAMGANRVIGNGPNIPWKIPGEQKIFRRLTEGKVVVMGRKTFESIGKPLPNRHTLVISRQANYRATGCVVVSTLSHAIALASELGNELYVAGGAEIYTLALPHAHGVFLSEVHQTFEGDAFFPMLNETEFELVSTETIQAVIPYTHSVYARRNG

>DHFR_salEnt Salmonella enterica
MNPESVRIYLVAAMGANRVIGNGPDIPWKIPGEQKIFRRLTESKVVVMGRKTFESIGKPLPNRHTVVLSRQARYSAPGCAVVSTLSQAIAIAAEHGKELYVAGGAEVYALALPHANGVFLSEVHQTFEGDAFFPVLNAAEFEVVSSETIQGTITYTHSVYARRNG

>DHFR_klePne Klebsiella pneumoniae
MNPELVRIYLVAAMGANRVIGNGPDIPWKIPGEQKIFRRLTEGKVVVMGRKTFESIGKPLPNRRTVVLSRQASYSAAGCAVVSTLSQAIAIAAEHGKELYVAGGAEVYALALPRADGVFLSEVHQTFEGDAFFPVLDEAEFEVVSAETVQATITYTHSVYARRNG

>DHFR_halNea Halothiobacillus neapolitanus
MNQQAVRIYLVAAMGSNRVIGNGSNIPWRIPGEQKIFRTLTEGKVVVMGRKTFESIGKPLPNRHTVVISRQTHYLAPGCTVATTLAQAIAIAAEAGKDLYVAGGAEIYALAMPHAHGIFLSEVHQTFDGDAFFPKLDEKDFELVSSETIQAAIPYTYSVYERRNG

>DHFR_macCas Macrococcus caseolyticus
MIISLIAAISSNYVIGKDKDIPWKIPGEQVRFKDLTMGKSVIMGRKTFESIGQPLPNRKTIIISKSKDINYNNCLTVESLERAFNLLQQEDEIFIAGGGEIYKESLPFADRIYLTIIEKEYEGNIFFPMFNKDEFEITYQQKVEGFIPYTYFTYERKNKGEQIK

>DHFR_geoUra Geobacter uraniireducens
MVISLIAAMAENRVIGRNNAIPWDIPADRKRFRALTLGHPVIMGRKTFESLAGPLPGRKNIIITRQMDYRAEGCFIAHDLASALAMAGETDEIFICGGGEIYRQALPIADRIYLTIIHSRFDGDTRFPEIPERFVEVGRETVENAIPYSFVLYDKRPSSASPA

>DHFR_breLat Brevibacillus laterosporus
MLSSIFAMGQNRVIGRDNQLPWRLPEDLKYFRRITTGHAIIMGRKTYESIGKPLPNRRNIVLTTQENYKADGCEIAHSIEEVLSLIDHKEEAFIIGGAEIYKLLLPFTSKMYITKIGQEFEGDAFFPAFDESEWALVEVTPGVQNEENPYQYEFQIYVRSVTNCK

>DHFR_oxaFor Oxalobacter formigenes
MMTISHVVAMAENRVIGKDGQMPWHIPGEQKIFRELTVGKALILGRKTHESIGRVLPDRITIIVSRKKDYEVPGAHVVHDIDEAIRLAREFGKEDIVIGGGGELFSQTLPVTDKVYLSIVHADFDGETFYPELPKEEFVEKARKEIDASIPYAFVEFERVKGK

>DHFR_nocSpp Nocardioides spp
MTPGGKRVVLVAAVARNGVIGDGPDIPWQLPGEQRLFKGLTWGHILVMGRATYDSIGRPLPGRTTIVLTRSPDWSAEGVHVAHDLPAALALADTLEGEVMVAGGGQVYAAALPVADEQVLSEVDLEPQGDAFYPEFDRSEWRETAREQHDGYDRVVLVRA

>DHFR_cloCel Clostridium cellulovorans
MISLIVAVAKNNVIGNNGIIPWKIKGEQKRFKELTIGKTIIMGRKSFEEIGKPLPNRKTILISNTQCIESENCTTVNSLSEAFDLTKDEDEVFVAGGGQVYKESFTYADRIYITVIDKIIDGNVYFPEISSDDFIKSYEERIDAEIPYTYYTYERRK

>DHFR_marPos Marinomonas posidonica
MLSLIVAMSTNRTIGINNSLPWHLPNDLKYFKQATMGKPIVMGRKTFESIGKPLPGRRNIVITRDANYQADGIDVVSSLEQAISLGEDICLVDGQEEVMVIGGAQIYELALPKADRLYITHVDAEVNGDAFFPEVDWPSFTVMAEESFAAEGPNPYDYRFSVYQRTSAE

>DHFR_pseAla Pseudoramibacter alactolyticus
MIALIAACDRHRLIGDHGRIPWRIPGEQARFQKLTTGHVVIMGRQTYAEIGRPLPRRQTIVLSRNQTFRAPGCRIAASLAEALRLGAAFSDQLFIAGGAAVYSKALPLADVLYLTEIDAAYVGDTYFPAFDPALFTKTLDAHISGDPSYDYYTYLRKS

>DHFR_halOch Haliangium ochraceum (bacteria) CP001804 Deltaproteobacteria MSSPSAPPNDTSS- trimmed
ASTPADAEHVFDLVVVADLDGGIARKGEIPWHLSADLRHFQQLTRTTAEPAQQNAVIMGRKTWESLPAASRPLPSRRNIVLSRDAELTLPPGVVHAAELEAALAATDLATPPIDRRFIIGGKGVYAEAMQRRGCRHIFYTEIQDRFDCDLFFPAFKDRFKRTDLLAEGEEKGIGYRIEVWTRTGFPGPV

>DHFR_braInt Brachyspira intermedia (bacteria) CP002874 Spirochaetes
MIVSLIAAVDSKNGIGLNGVMPWGHIKEDMQFFRSTTTGYAVVMGRVTFESLGSKPLPNRKNIVISSSINNDLLEKYDNLFYESSFENTISKLLLEKNNQIFIIGGESIYKKALDYADIIYLTHIDKDYNCDRFFPQIDTKLFQSSKLKTFFHNDININIIKYTRTLL

Duplicated DHFRL1 in primates

This gene is a spin-off of the parent DHFR gene that occured in stem primates. As a retrogene, it has lost all introns. In some species including human, it has been reported to be functional though it is unclear how mitochondria accomplished this function prior to the duplication. In other species, this locus has recently pseudogenized leaving only debris; here some function must have persisted for millions of years in view of maintenance by selective pressure. DHFR, being ubiquitously expressed, has given rise to many other retroposed loci; some of these too may be functional.

>DHFRL1_homSap Homo sapiens (human) NM_001195643 synteny +ARL13B -DHFRL1 +NSUN3 chr3
MFLLLNCIVAVSQNMGIGKNGDLPRPPLRNEFRYFQRMTTTSSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINLVLSRELKEPPQGAHFLARSLDDALKLTERPELANKVDMIWIVGGSSVYKEAMNHLGHLKLFVTRIMQDFESDTFFSEIDLEKYKLLPEYPGVLSDVQEGKHIKYKFEVCEKDD

>DHFRL1_panTro Pan troglodytes (chimp) ti|1986883368 +ARL13B -DHFRL1 +NSUN3  short c-terminal
MFLLLNCIVAVSQNMGIGKNGDLPRPPLRNEFRYFQRMTTTSSVEGKQNLVIMGRKTWFSIPEKNRPLNDRINLVLSRELKEPPQGAHFFARSLDDALKLTERPELANKVDMIWIVGGSSVYKEAMNHLGHLKLFVTRIMQDFESDTFFSEIDLEKYKLLPEYPGVLSDVQEGKHIKYKFEVYEND

>DHFRL1_gorGor Gorilla gorilla (gorilla) ti|2051780408/ti|2033585870 10 bp deletion so frameshift; synteny unknown but 97% identical homSap
MFLLLNCIVAVSQNMGIGKNGDLPRPPLRNEFRYFQRMTTTSSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINLVLSRELKEPPQGAHFLAISLDDALKLTERPKLANKVDMIWIVGGSSVYKEAMNh lKLFVTRIMQDFASDTFFSEIDLEKYKLLPEYPGVLSDVQEgKHIKYKFEAYEKND
 
>DHFRL1_ponAbe Pongo abelii (orangutan) genome/blat +ARL13B -DHFRL1 +NSUN3  
MVRLLNCIVAVSQNMGIGKNGDLPWPPLRNEFRYFQRMTTTSSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINLVLSRELKEPPQGAHFLARSLDDALKLTERPELANKVDMIWIVGGSSVYKEAMNHLGHLKLFVTRIMQDFESDTFFSEIDLEKYKLLPEYPGVLSDVQEEKGIKYKFEVYEKND

>DHFRL1_nomLeu Nomascus leucogenys (gibbon) ADFV01005477 +ARL13B -DHFRL1 +NSUN3  
MVRLLNCIVAVSQNMGIGKNGDLPWPPLRNEFRYFQRMTTTSSVEGKQNLVIMGRKTWFSIPEKNRPLEDRINLVLSRELKEPPQGAHFLARSLDDALKLTEQPELANKVDMIWIVGGSSVYKEAMNRSGHLKLFVTRIMQDFESDMFFSEIDLEKYKLLPEYPGVLSDVQEEKGIKYKFEVYEKND

>DHFRL1_macMul Macaca mulatta (rhesus) genome/blat +ARL13B -DHFRL1 +NSUN3 
MVRSLNCIVAVSQNMGIRKNGDLPWPPLRNEFRYFQRMTTTSSVEGKQNLVIMGKKTWFSIPEKNRPLKGRINLVLSRELKEPPQGAHFLARSLDDALKLTEQPELANKVDMIWIVGGSSVYKEAMNHPGHLKLFVTRIMQDFESDTFLPEIDLEKYKLLPEYPGVLSDVQEEKGIKYKFEVYEKND

>DHFRL1_papHam Papio hamadryas (baboon) ti|1986883368 synteny unknown 
MVHSLNCIVAVSQNMGIGKNGDLPWPPLRNEFGYFQRMTTTSSVEGKQNLVIMGKKTWFSIPEKNRPLKGRINLVLSRELKEPPQGAHFLARSLDDALKLTEQPELANKVDMIWIVGGSSVYKEAMNHPGHLKLFLTRIMQDFESDTFLPEIDLEKYKLLPEYPGVLSDVQEEKGIKYKFEVYEKN

>DHFRL1_calJac Callithrix jacchus (marmoset) genome/blat frameshift and stop codon; +ARL13B -DHFRL1 +NSUN3 
mvrllCIVAVSQNMGIGKKGDLPWPPLRNEFRYFQRMTTTSSIEGKQNLVIMGRKTWFSIPEKNRPLKDRINFVLSRELQEPPQGAHFLARSLDDALNLTEQPELANKVDMIWIVGGSSVYKEAMNRPGHLKLFVTRIMQDFESDTFFPEIDLEK*KLLPEYPGVLSDVQEEKGIKYKFEVYEKND

>DHFRL?_otogar Otolemur garnettii (bushbaby) AAQR03025778 synteny unknown
MVHRLNCIIAVSQNRGIGKNGDLPWPPLRNEFNYFQRMTTTSSVEGKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKESSQGAHFLAKSLDDALKHIEEPELANKVDMVWIVGGSSVYKEAMNQPGHVKLFVTRIMQDFECDTFFPEIDLKKYKLLSEYPGVLSDVQEEKGIKYRFEVYEKND

>DHFRL?_micMur Microcebus murinus (lemur) ABDC01288917 synteny unknown 
MVRTLNCIVAVSQNMGIGKNGDLPWPPLRNEFRYFQKMTTTSSVEGKQNLVIMGRKTWFSIPEKSRPLKDRINVVLSRELKEPPQGAHFVAKSLDDALKLIEQPELANKVDMVWIVGGSAVYKEAMNQPGHLKLFVTRIMQDFESDTFFPEIDLKKYKLLPEYPGVLSDVQEEKGIKYKFEVYEKND

>DHFRL??_micMur Microcebus murinus (lemur) ABDC01470170 synteny unknown two stop codons and a frameshift
MVHTLNCLVAVSQNMGIGKNGDLPWPPLRNEFKYFQKMTTTSSVEGKQNLVIMGKKTWFSIPEKN*PLKDRINVVLSREL*EPPQGAHFIAKSLDDALKLIEQPELANKVDMVWIVGGSAVYKEAKNQPGHLKLFVTRIMQDFESDTFFPEIDMKKYKLLSECPGVLSDVQEEKGMKYKFEVYERnd

>DHFRL?_tupBel Tupaia belangeri (treeshrew) AAPY01242529 several candidates but synteny unknown retrogene shown two frameshifts
MVRPLDCIVAVSQmGIGKNGDLPWPPLRNEFKYFQRMTTTSSVEGKQNTVIMGRKTWFSIPEKNRPLKDRINLVLSRELKEPPRGAHFLAKSLDDALKLIEQPELANQVDMVWIAGGGSVYKEAMNQPGHLKPFVTRIMQDFESDTFfPEIDLEKYKLLPGYPGVLSDDQEEKGIKYKFEVYEEN

Duplicated DHFRL2 in bony vertebrates

This locus is a small segmental duplication of the parent DHFR gene that occured in stem teleosts and persisted through lobe-finned fish and amphibian divergences, only to be lost in all amniotes including human. It has retained all introns and splice junction phases. Ray-finned fish supposedly had a stem whole genome duplication -- had both copies been retained, three diverged copies of DHFR in paralogous blocks would be present today. Coelocanth, a lobe-finned fish, and frog were not subject to this whole genome duplication.

Zebrafish and other ray-finned fish have a second functional copy of DHFR denoted here as DHFRL2. It did not arise from whole genome duplication, because the two loci are utterly different in syntenic context and since the duplicated locus also occurs in coelocanth and two amphibians. The gene tree for DHFRL2 forms a distinct, more diverged cluster that branches off from DHFR in early bony vertebrates.

This feature cannot be a pseudogene because of billions of years of observed branch length conservation. Pseudogenes persist for a few million years at most so quickly become undetectable. DHFRL2 could conceivably be a very recent pseudogene in some species but not all because that requires many separate events which is not parsimonious. DHFRL2 orthologs are currently available for 15 species.

However its function is obscure: it may function in mitochondria like primate DHFRL1, or be specialized to expression in a developmental stage or intra-cellular location, or have a slightly different substrate, or even have a non-catalytic regulatory role.

The main DHFR has syntenic context FAM151B ANKRD34B DHFR MSH3 RASGRF2 whereas the secondary DHFR2 sits in completely different surroundings RFX4 RIC8B SIRT3 C12orf23 CHST1 MTERFD3 DHFRL2 CRY1 BTBD11. Frog, like all rayfinned fish, unmistakably has a DHFR in the first spot and a DHFRL2 in the second. For a whole genome duplication, the two blocks need to be paralogous, here they are not. Note syntenic context for the primate retroposed gene DHFRL1 has, as expected, no relation to either site: ARL13B DHFRL1 NSUN3.)

Thus the duplication event occurred in the last common ancestor of amphibians and ray-finned fish -- the stem bony vertebrate thus had both DHFR and DHFRL2 at these two sites in the ancestral chromosomes. The two genes have persisted some 450 myr to contemporary amphibians, coelocanth and ray-finned fish but the DHFRL2 gene was lost in the amniote stem with unknown consequences.

The gene duplication may be slightly older but assemblies are inadequate in chondrichthyes, lamprey and cephalochordate. The tunicate Ciona has a close anti-parallel tandem duplication of DHFR in an altogether novel syntenic location (tunicate gene order is typically scrambled).

A paralog of DHFR in coelocanth cannot be confirmed as DHFRL2 by synteny because the contigs are too small. However it clusters as expected in the combined gene tree. As expected for a 'living fossil', both DHFR and DHFRL2 have evolved slower than other species. Thus on Blastp query against human, both proteins 'jump the queue' (score higher than later diverging species). However this could be due to more rapid evolution in the other species.

The alignment below compares separately determined conservation in species-matched sets of 15 DHFRL2 and DHFR sequences. While DHFR is significantly more conserved, DHFRL2 also has very significant conservation (shown below at the 90% level). Although certain residues are conserved between both, each of the proteins has a considerable number of conserved residues not shared.

DHFRL2_xenTro KPVKLIAAACNSMGIGLNGYLPWN---LPNEFKYLLDTITRVTQPGKKNLLLWGRQSFETFDESLLPLPNCIIALLSRKLSTVPPYVSYLCKNEEEILKFASTPPLNDEIETIWVLGGVESYKNLMQHPWCNHIYFTKIMADFECDTFFPEFDKNIFKLKENFPGVPSGIQEENGVKYVFQVYQR
DHFRL2_ambTig KPIKIIVAAGRNRGIGIKGNLPWN---LPNEFKYFLDKITSVSNPGKKNFIMVGRTSFEGFPDCVLPLPNTIVAILSKTLRSTPKHAAYVCQDVQEVLDLASSPPLSKDIETIWALGGVQSYMEVMHHPWCNQIFFTNVMADFECDTFFPAIDTGVFNLVEQFPGVPLGLQEENGIKYEFQVYER
DHFRL2_latCha KPICLIVAACKHMGIGKGGDLPWH---LPNEFKHFLKMTTTVSVPGKKNLLVSGRKSYEPFPKSCMPLPNCYIAVLSRTLKSVPKHAHYLCRDLPSAIKLASTPPLSKEIETIWIIGGVETYIEA-KHPWCELLYLTEIMAEFDCDTFFPEFDRKVYKLVDEFPGVPTGIQEEEGIKYKFHVFKR
DHFRL2_danRer KPIRLIAAACRDMGIGKDGQIPWC---LPKEFQFLLDTITAVSAPGKKNLIVWGRICWFSCPETVFPLANCINLVLSRKMISVPPHAHYLCKDFDSIIRLVSEPPLCHTVEVIWILGGTEVYKESLEHPWCDLIYLTDIMANFECDVFFPEFDPNIFRKQKSFPGVPDEIIEENGIKFQFQVFK-
DHFRL2_pimPro KPIRLIAAACRDMGIGKNGQLPWS---LPKEFQFFLDTITAVSTPGKKNLVVWGRVSWFSCPETVFPLANCFNLVLSRKLNANPPRAHYLCKDFESVIRLVSEPPLCHIVETIWVLGGPEVYKESLEHPWCDRIYLTDIMADFDCDVFFPKFDCNIFRKQKGFPGVPDEIHEENGIKFQFQVFKR
DHFRL2_ictPun KPIRLVAAACRNMGIGKDGRLPWN---LPTEFQFFLNTITAVCKSGKKNLIIWGKYSWFSCPESVFPLANSLHVVLSKKLMSVPKRAHYVCEDFVSAVKLASHPPLNDLIETIWILGGTQVYKEALEHPWCDLIYLTDIMADFDCNVFFPSFDQKFYRKQNKFPGVPNEIQEENGIKFRFQVFMK
DHFRL2_tetNig KPVRLIAAKCNGGGIGKDNRMPWS---LPSEFRFFLSTITRVSRPGKMNMIIWGKQCWISHPESTFPLPNVLHTVLSRTLFAVPDHAHFVCKTLDAAVRLASQPPLADLIEVVWIVGGVQVYKEAMEHPWCDLIYLTDIMAEFECDVFFPEFDRQLFQVQDGFPDVPDGIQEENGIKYKCQVYKR
DHFRL2_oncMyk KPVRLIAAACNNMGIGKDGHLPWD---LPTEFKFFLETTTNVSLPGKFNMMVWGRGCWFSNPDSFFSLPNVLHVVLSTTLSTLPEHAHFLCQDFDSMISLALLQPLCDLVETIWVVGGPRVYQEALMHPWCELVFLTDIMADFDCDVFFPQFDRSVFKKQQRFPGVPNEMQEENGVTFKYEVFKR
DHFRL2_takRub KPVRLIAAKCNGGGIGKDNRMPWS---LPAEFQYFLNTVTRVSRPGNMNMMVWGKQCWVSHPDSTFPLPNILHAVLSKTLLTVPDHAHFLCESLDAAVRLASEPPLADLIEIIWIVGGVQVYKEAMEHPRCDLIYLTDIMAEFECDVFFPEFDKKLFEVQDRFPDVPNGIQEDNGIKYKCQVYKR
DHFRL2_oreNil KPVRLIAAACNDMGIGKDGKLPWD---LPTEFQYFVNTVKTVSRPGRMNFMVWGRLCWDSHPESLFPLPNILHVVLSKTLDTVPDHAHFLCRDFDSAIRLAAEPPLADLIETIWIVGGVQVYKDALNHPWCDLLYLTDVMADFDCDVFFPEFDKNLFQLQEGFPGVPSEIQEENGIKFKYQVFKR
DHFRL2_hipHip KPVRLIAAACNDMGIGKDGKLPWK---LPSEFQYFLNTVTRVSRPGKMNMMVWGKLCWFSHPETMFPLANTLHVVLSTKLNKVPDHAHFLSEDFESAVRMSAEPPLADLIENIWVVGGMQVYKDALSHPWCDLVYLTNVMADFECDVFFPEFDRGVFKTQKDFLEVPSGIQEEDGIKYKFQVFKK
DHFRL2_poeRet KPVRLIAANCNNMGMGKDGTMLWS---LPSEFQYFMNTITRVSRPGNMNLLIWGRLCWNSHPENIFPLANTLHAVLSKTSSSVPDHAHFLCQDFESAVRLAAQPPLSGIIETVWILGGTQVYEDALKHPWCDLLYLTDVMADFDCDVFFPDFDRELFKLQEAFPDVPSEVQEENGIRFKCQVFKK
DHFRL2_gasAcu KPVRLIAAACSGGGIGKDGQMPWD---LPSEFRYFRNHVQGVSRPGKMNMMVWGKLCWYSVPKPDFPLPNVLHVVLSKTLESVPDHAHFLCEDLDAAARLAVQPPLADLIETIWVVGGTQVYEVALKHPWCDLVYLTDVMADFDCNVFFPEFDRGLFKVQEGFPGVPSEIQEENGIKYKFQVFKK
DHFRL2_dicLab KPIRLIAVVCNGNGIAKDGKMPWN---LPSEFQFFLKTVTTVSRPGKMNFLVWGKRTWQSHPETTLPLANTLHVVLSTELKTVPDHAHFLSQDFESAVRLASQPPLADLIETIWIIGGTQVYEDALKHPWCDLVYLTDVMADFECNVFFPEFDRGLFKLLENFPGVPSGIQEENGTKYKCQVFKR
DHFRL2_gadMor KPVRLIAAACNDLGIGKNGKLPWS---LPNEFRYYLNNLSRVSRPGMFNLVVWGRLNWFSQEE--FLKENVILVVLSKTLGVVPDNAQYLREDLEGAIRLAASPPLSDIIETIWVVGGTGVYKESLMHPWCDLIYLTDVMANIDCDVFFPNFDRQIFKKQDRFPGVPSEIQEENGVKYKYEVYKK

DHFR_homSap   GSLNCIVAVSQNMGIGKNGDLPWPPLR--NEFRYFQRMTTTSSVEGKQNLVIMGKKTWFSIPEKNRPLKGRINLVLSRELKEPPQGAHFLSRSLDDALKLTEQPELANKVDMVWIVGGSSVYKEAMNHPGHLKLFVTRIMQDFESDTFFPEIDLEKYKLLPEYPGVLSDVQEEKGIKYKFEVYEK
DHFR_xenTro   PFLHAVVAVCPNQGIGKEGSLPWPLLR--NEFKHFQRLTMTPTVEDKKNVVIMGRKTWFSIPEKNRPLKERINIVLSKELKEPPTGAHYLSKSLAEAIDLLEDPELKDKVDLVWVIGGSSLYQELMEKPVNQRWFVTRILQEFECDTYLPEIDLNSFRLLPEYPGISPELQEENGVQYKFEVYEK
DHFR_ambMex   RILNAIVAVCPSMGIGKDGNLPWPILR--NEFKYFQRMTMTATQEGKQNVVIMGRKTWLSIPEKNRPLKGRINIVLSRELTAPPPGAHYLANSLDGALELLETRELAGKVDLLWIIGGSSLYRELIGRPHHQRLFVTRILKEFECDTFLPEFDLEKYRLLPEYPGVSLGIQEENGIQYKFEVYEK
DHFR_latCha   RLLNSIVAVCPNLGIGKDGNLPWHPKRLSNEFRYFQKMTTTPTVEGKQNVVIMGRKTWFSIPEKNRPLKGRVNVVLSRELQEAPEGAHYLARDLDSALKLLESPELADRVDLVWIIGGSALYKEALEHPSQHRLFVTRVLQQFECDTYLPEINFNKFTLLPEFPGVSSAIQEESGIQYKFEVYEK
DHFR_danRer   RILNCIVAVCPDMGIGKNGNLPWHPIRLSNELKHFQKMTMTPSDEGKKNVVIMGRKTWFSIPAAHRPLKNRINIVLSRELKTAPEGAHYLASDFSSALHLLDSGELEKLVDQVWIIGGSSLYKEVMERSGHRRLFVTRILKQFDCDTFIPNFDMDKYKLLPEFPGVPVGLQEDNGVQYLFEVYES
DHFR_cypCar   RILNSIVAVCPDMGIGKNGNLPWHPIRLSNEFKHFQKMTMTPLVEGKKNVVLMGRKTWFSIPAANRPLKNRINIVLSRELKTAPEGAHYLASDFSSALHLLDSSELEKQVDQVWIIGGSALYKEVMESSGPRRLFVTRVLKQFDCDTFIPDINMDKYKLLPEFPGVPTGLQEENGVQYVFEVYES
DHFR_ictPun   RVLNCIVAVCPDMGIGRNGNLPWHPIRLSKEFKHFQKMTMTPTVEGKKNVVIMGRKTWFSIPAQNRPLKNRINVVLSRELKTAPEGAHYLAADFSSALRILENAELEAQADQVWIIGGSSLYKEVMESSAHQRLFVTRILKQFDCDTFIPNIDLDKFRLLPEFPGVPAGLQEENGLQYVFEVYES
DHFR_oncMyk   RVLNCIVAVCPDMGIGNNGNLPWHPKRLNNEFKYFQKMTMTSSVEGKQNAVIMGRKTWFSIPERNRPLKNRINIVLSRELKEPPAGAHQLASDFRSALRLLDTAEMAERVDQVWVIGGSSLYEEMMESQGTRRLFVTRILQQFKSDTFLPEINQDKYRLLPEFPGVPQDLQEEAGLQYRFEVYES
DHFR_esoLuc   RVLNCIVAVCPNMGIGNKGNLPWHPKRLNNEFKYFQKMTMTPFVEGKQNVDIMGRKTWFSIPERNRPLKNRINIVLSRELKEPPPGAHHLASDFSSALRLLDTAEMAERVDQVWVIGGSSLYKEMMESEGIRRLFVTRILQQFESDTFLPEINQDKYRLLPEFPGVPQDVQEESGIQYRFEVYES
DHFR_gadMor   RVLNCIVAVCPDAGIGYKGDLPWHPTRLNNEFKHFRRLTVTPGAEDKQNVVIMGRKTWNSIPEKNRPLNDRINIVLSRQLEAPPSGAHHLARDLPSALRLLDV-PEGLLADQVWLIGGSSMYSEVMETPCPRRLFVTRILKQFPSDTFFPDISPAQFRLLPEYPGVSSELQEENGIQYRFQVYQS
DHFR_tetNig   RVLNAIVAVCPDLGIGRNGDLPWHPIRLDNEFKHFRKMTSTPSVNGKQNVVIMGRKTWFSIPEKHRPLANRINIVLSRRSREPPAGAHHLAHDLSSALQLVET-QLAEQADQVWVIGGSSLYQEMMERPGTSRLFVTHVLKQFDCDTFLPEIRPQRYRLLPQFPGVPQELQEEKGIQYRYQVYES
DHFR_oreNil   RVLNAIVAVCPDRGIGNKGNLPWHPIRLSKEFAHFRKMTATPSVKGKQNVVIMGKKTWYSIPEKNRPLSNRINIVLSRECKVPPAGAHYLASDFSSALRLIDT-ELADQADQVWVIGGSSLYKEMMGSTGMRRLFVTQILKQFECDTFLPEISLDKYRLLPEFPDVPQELQEENGIQYRFKVYES
DHFR_hipHip   RILNGIVAVCPDLGIGNRGNLPWHPVRLSNEFKHFRSMTATPSEKGKQNVVIMGRKTWFSIPEKNRPLNNRINIVLSRQCKAPPAGAHHLSADFSSALRLVDT-ELADQADQVWVIGGSSLYKEMMESPGTRRLFVTRILKQFDCDTFLPEISPDKYRQLPEFPGVPQELREENGIQYRFEVYES
DHFR_gasAcu   RVLNGIVAVCPDLGIGCHGNLPWHPLRLSNEFKHFRRMTATASVKDKQNVVIMGRKTWFSIPEKNRPLNNRINIVLSRELKAPPAGAHHLAPDFSSALRLVDT-ELAEQADQVWVIGGSSLYKELLESPGTRRLFVTRILKQFECDTFLPEICPDRYRLLPEFPGVPEELQEENGVQYRFEVYES
DHFR_dicLab   RILNGIVAVCPDLGIGMNGNLPWHPVRLGNEFKHFRTMTATPSVKGKQNVVIMGRKTWFSIPEKNRPLNNRINIVLSKECKVPPAGAHHLAADFNSALRLVET-ELADQADQVWVIGGSSLYKELMESPGTRRLFVTRILKQFECDTFLPEICPDKYQLLPEFPGVSQELQEENGIQYRFEVYES
DHFR_spaAur   RIVNGIVAVCPDLGIGNNGNLPWHPVRLNREFKHFRTMTATPSVKGKQNVVIMGRKTWFSIPDKFRPLNNRINIVLSRECKVPPAGAHHLAADFSSALSLVDT-ELADQADQVWVIGGSSLYKELMESTRTSRLFVTRILKQFECDTFLPEISPDKYSLLPEFPGVPQGLQEENGIQYKFEVYER

DHFRL2_latCha KPICLIVAACKHMGIGKGGDLPWH---LPNEFKHFLKMTTTVSVPGKKNLLVSGRKSYEPFPKSCMPLPNCYIAVLSRTLKSVPKHAHYLCRDLPSAIKLASTPPLSKEIETIWIIGGVETYIEA-KHPWCELLYLTEIMAEFDCDTFFPEFDRKVYKLVDEFPGVPTGIQEEEGIKYKFHVFKR
90% conserv   KP..LI.A.C...GIG.....PW.---LP.EF.........V..PG..N....G.............L.N.....LS......P..A................PPL....E..W..GG...Y.....HPWC.....T..MA.F.C..FFP.FD.........FP.VP....EE.G......V...
unique cons   KP..L...................---LP............V..PG.......................N.................................PpL....e................HPWC........mA...C..FF..FD.........F..V...................
shared cons   .....I.A.C...GIG.....PW.---...EF................N....G.............L.......LS......P..A.................2.....2..W..GG...Y.................2.........P.............P.V.....EE.G......V...
unique cons   ..LN..V.V.P.......G.L.....R.......F...T.T.....K...VIM..KTW.SIP...RP...RIN.V..........G.H.L......AL.L....e.....d.V..I..S....E........RLFV...L.....DT...........LLPE..G.....Q.....QY.F..YE.
90% conserv   ..LN.IVAVCP..GIG..G.LPW...R...EF..F...T.T.....K.N.VIMG.KTW.SIP...RPL..RIN.VLS......P.GAH.L......AL.L....E.....D.VW.IGGS..Y.E........RLFVT..L..F..DT..P........LLPE.PGV....QEE.G.QY.F.VYE.
DHFR_homSap   GSLNCIVAVSQNMGIGKNGDLPWPPLR--NEFRYFQRMTTTSSVEGKQNLVIMGKKTWFSIPEKNRPLKGRINLVLSRELKEPPQGAHFLSRSLDDALKLTEQPELANKVDMVWIVGGSSVYKEAMNHPGHLKLFVTRIMQDFESDTFFPEIDLEKYKLLPEYPGVLSDVQEEKGIKYKFEVYEK

>DHFRL2_xenTro Xenopus tropicalis (frog)  BTBD11 RASGRF2 MSH3 DHFRL2 CRY1 MTERFD3 SIRT2 RIC8A RFX4 on truncated contig chr GL173571, dozens of transcripts
MREAPNGTKPVKLIAAACNSMGIGLNGYLPWNLPNEFKYLLDTITRVTQPGKKNLLLWGRQSFETFDESLLPLPNCIIALLSRKLSTVPPYVSYLCKNEEEILKFASTPPLNDEIETIWVLGGVESYKNLMQHPWCNHIYFTKIMADFECDTFFPEFDKNIFKLKENFPGVPSGIQEENGVKYVFQVYQRDLLVDLL*

>DHFRL2_ambTig Ambystoma tigrinum (salamander) CN067598 no synteny available
MSEARERNGPKPIKIIVAAGRNRGIGIKGNLPWNLPNEFKYFLDKITSVSNPGKKNFIMVGRTSFEGFPDCVLPLPNTIVAILSKTLRSTPKHAAYVCQDVQEVLDLASSPPLSKDIETIWALGGVQSYMEVMHHPWCNQIFFTNVMADFECDTFFPAIDTGVFNLVEQFPGVPLGLQEENGIKYEFQVYERKGSISM*

>DHFRL2_latCha Latimeria chalumnae (coelocanth) AFYH01018047 AFYH01018044 AFYH01018043 AFYH01018041 second exon missing, filled by consensus
MGIGKKPICLIVAACKHMGIGKGGDLPWHLPnefkhflkmtttvsvpGKKNLLVSGRKSYEPFPKSCMPLPNCYIAVLSRTLKSVPKHAHYLCRDLPSAIKLASTPPLSKEIETIWIIGGVETYIEAKHPWCELLYLTEIMAEFDCDTFFPEFDRKVYKLVDEFPGVPTGIQEEEGIKYKFHVFKRD

>DHFRL2_danRer Danio rerio BC165261  BTBD11 MSH3 CRY1 DHFRL2 MTERFD3 SIRT2 RIC8A RFX4
MNIEGEEMRKPIRLIAAACRDMGIGKDGQIPWCLPKEFQFLLDTITAVSAPGKKNLIVWGRICWFSCPETVFPLANCINLVLSRKMISVPPHAHYLCKDFDSIIRLVSEPPLCHTVEVIWILGGTEVYKESLEHPWCDLIYLTDIMANFECDVFFPEFDPNIFRKQKSFPGVPDEIIEENGIKFQFQVFKKIKN*

>DHFRL2_pimPro Pimephales promelas (fathead minnow) DT166997
MNIEGEEMRKPIRLIAAACRDMGIGKNGQLPWSLPKEFQFFLDTITAVSTPGKKNLVVWGRVSWFSCPETVFPLANCFNLVLSRKLNANPPRAHYLCKDFESVIRLVSEPPLCHIVETIWVLGGPEVYKESLEHPWCDRIYLTDIMADFDCDVFFPKFDCNIFRKQKGFPGVPDEIHEENGIKFQFQVFKrA*

>DHFRL2_ictPun Ictalurus punctatus (catfish) FD330522 
MKPRNEVQKPIRLVAAACRNMGIGKDGRLPWNLPTEFQFFLNTITAVCKSGKKNLIIWGKYSWFSCPESVFPLANSLHVVLSKKLMSVPKRAHYVCEDFVSAVKLASHPPLNDLIETIWILGGTQVYKEALEHPWCDLIYLTDIMADFDCNVFFPSFDQKFYRKQNKFPGVPNEIQEENGIKFRFQVFMKES

>DHFRL2_tetNig Tetraodon nigroviridis (pufferfish) CR727807 cDNA
MEPAKVIKKPVRLIAAKCNGGGIGKDNRMPWSLPSEFRFFLSTITRVSRPGKMNMIIWGKQCWISHPESTFPLPNVLHTVLSRTLFAVPDHAHFVCKTLDAAVRLASQPPLADLIEVVWIVGGVQVYKEAMEHPWCDLIYLTDIMAEFECDVFFPEFDRQLFQVQDGFPDVPDGIQEENGIKYKCQVYKRERAAEDQQ*

>DHFRL2_oncMyk Oncorhynchus mykiss (rainbow trout) CX259371 fragment
qkkpvrlIAAACNNMGIGKDGHLPWDLPTEFKFFLETTTNVSLPGKFNMMVWGRGCWFSNPDSFFSmLPNVLHVVLSTTLSTLPEHAHFLCQDFDSMISLALLQPLCDLVETIWVVGGPRVYQEALMHPWCELVFLTDIMADFDCDVFFPQFDRSVFKKQQRFPGVPNEMQEENGVTFKYEVFKREICSAEVI*

>DHFRL2_takRub Takifugu rubripes (fugu) CAAB02000170
MEPAKVIKKPVRLIAAKCNGGGIGKDNRMPWSLPAEFQYFLNTVTRVSRPGNMNMMVWGKQCWVSHPDSTFPLPNILHAVLSKTLLTVPDHAHFLCESLDAAVRLASEPPLADLIEIIWIVGGVQVYKEAMEHPRCDLIYLTDIMAEFECDVFFPEFDKKLFEVQDRFPDVPNGIQEDNGIKYKCQVYKRKTAGKDQQ*

>DHFRL2_oreNil Oreochromis niloticus (tilapia) XM_003447073 dihydrofolate reductase-like
METSWEKLQKKPVRLIAAACNDMGIGKDGKLPWDLPTEFQYFVNTVKTVSRPGRMNFMVWGRLCWDSHPESLFPLPNILHVVLSKTLDTVPDHAHFLCRDFDSAIRLAAEPPLADLIETIWIVGGVQVYKDALNHPWCDLLYLTDVMADFDCDVFFPEFDKNLFQLQEGFPGVPSEIQEENGIKFKYQVFKRDTADTV*

>DHFRL2_hipHip Hippoglossus hippoglossus (Atlantic halibut) EB034565 frameshift fixed
MEKSREKRQKKPVRLIAAACNDMGIGKDGKLPWKLPSEFQYFLNTVTRVSRPGKMNMMVWGKLCWFSHPETMFPLANTLHVVLSTKLNKVPDHAHFLSEDFESAVRMSAEPPLADLIENIWVVGGMQVYKDALSHPWCDLVYLTNVMADFECDVFFPEFDRGVFKTQKDFLEVPSGIQEEDGIKYKFQVFKKSICDAI*

>DHFRL2_poeRet Poecilia reticulata (guppy) ES386580
MEKDLEKVQKKPVRLIAANCNNMGMGKDGTMLWSLPSEFQYFMNTITRVSRPGNMNLLIWGRLCWNSHPENIFPLANTLHAVLSKTSSSVPDHAHFLCQDFESAVRLAAQPPLSGIIETVWILGGTQVYEDALKHPWCDLLYLTDVMADFDCDVFFPDFDRELFKLQEAFPDVPSEVQEENGIRFKCQVFKKKTDDTM

>DHFRL2_gasAcu Gasterosteus aculeatus (stickleback) ENSGACT00000017858 
MSGDNVQKKPVRLIAAACSGGGIGKDGQMPWDLPSEFRYFRNHVQGVSRPGKMNMMVWGKLCWYSVPKPDFPLPNVLHVVLSKTLESVPDHAHFLCEDLDAAARLAVQPPLADLIETIWVVGGTQVYEVALKHPWCDLVYLTDVMADFDCNVFFPEFDRGLFKVQEGFPGVPSEIQEENGIKYKFQVFKKETGDAV

>DHFRL2_dicLab Dicentrarchus labrax (seabass) FM022412
METVQRKPIRLIAVVCNGNGIAKDGKMPWNLPSEFQFFLKTVTTVSRPGKMNFLVWGKRTWQSHPETTLPLANTLHVVLSTELKTVPDHAHFLSQDFESAVRLASQPPLADLIETIWIIGGTQVYEDALKHPWCDLVYLTDVMADFECNVFFPEFDRGLFKLLENFPGVPSGIQEENGTKYKCQVFKRETGDTV

>DHFRL2_gadMor Gadus morhua (Atlantic cod) ES776457
MSLQKKPVRLIAAACNDLGIGKNGKLPWSLPNEFRYYLNNLSRVSRPGMFNLVVWGRLNWFSQEEFL  KENVILVVLSKTLGVVPDNAQYLREDLEGAIRLAASPPLSDIIETIWVVGGTGVYKESLMHPWCDLIYLTDVMANIDCDVFFPNFDRQIFKKQDRFPGVPSEIQEENGVKYKYEVYKKDIHHQDKLM*

Blastp scores relative to human DHFR
DHFR_latCha   Latimeria chalumnae (coelocanth) AFYH011508...   709  1.9e-74
DHFR_xenTro   Xenopus tropicalis (frog) CT025223  RASGFRA...   653  1.5e-68     
DHFR_ictPun   Ictalurus punctatus (catfish) FD330522           613  2.6e-64     
DHFR_danRer   Danio rerio (zebrafish) NM_131775 CELA2A BC...   607  1.1e-63     
DHFR_gasAcu   Gasterosteus aculeatus (stickleback) ENSGAC...   600  6.3e-63     
DHFR_tetNig   Tetraodon nigroviridis (pufferfish) CR710339     596  1.7e-62     
DHFR_oreNil   Oreochromis niloticus (tilapia) GR620119         583  4.0e-61  
   
DHFRL2_latCha Latimeria chalumnae (coelocanth) AFYH010180...   453  1.4e-47
DHFRL2_oreNil Oreochromis niloticus XM_003447073 dihydrof...   418  7.1e-44
DHFRL2_danRer Danio rerio BC165261  BTBD11 MSH3 CRY1 DHFR...   403  4.7e-42 Identity: 75/181 (41%) Similar: 115/181 (63%)   
DHFRL2_gasAcu Gasterosteus aculeatus (stickleback) ENSGAC...   414  3.2e-43     
DHFRL2_ictPun Ictalurus punctatus (catfish) FD330522           406  2.3e-42     
DHFRL2_tetNig Tetraodon nigroviridis (pufferfish) CR72780...   404  3.7e-42 
DHFRL2_xenTro Xenopus tropicalis (frog)  BTBD11 RASGRF2 M...   366  3.9e-38 Identity: 72/179 (40%) Similar: 108/179 (60%) 

Supporting syntenic data:
Brown: definite DHFRL2; Green: no data but DHFRL2 expected; Red: DHFRL2 lost

Gnathostomata (jawed vertebrates)
    Chondrichthyes (cartilaginous fish)
    Teleostomi (bony vertebrates)
        Actinopterygii (ray-finned fishes)
            Actinopteri
                Chondrostei (sturgeons)
                Neopterygii
            Polypteriformes
                Polypteridae (bichirs)
        Sarcopterygii
            Coelacanthiformes (lobe-finned fishes)
            Dipnoi (lungfishes)
            Tetrapoda (tetrapods)
                Amniota (amniotes)
                Amphibia (amphibians)

Gene order in Homo sapiens (human):
  FAM151B          800     9   276   276 100.0%     5  ++   79797610  79837648  40039
  ANKRD34B        1542     1   514   514 100.0%     5  +-   79854297  79855838   1542
  DHFR             556     1   187   187 100.0%     5  +-   79924909  79950308  25400
  MSH3            3388     1  1137  1137 100.0%     5  ++   79950547  80171678 221132
  RASGRF2         3675     1  1237  1237  99.9%     5  ++   80256558  80521586 265029

  RFX4            2203     1   744   744  99.9%    12  ++  106995055 107155244 160190
  RIC8B           1552     1   520   520 100.0%    12  ++  107168524 107279790 111267
  [SIRT3]
  C12orf23         347     1   116   116 100.0%    12  ++  107360895 107365166   4272
  [CHST1]
  MTERFD3         1155     1   385   385 100.0%    12  +-  107371338 107372492   1155
  [DHFRL2]
  CRY1            1747     1   586   586 100.0%    12  +-  107386568 107486739 100172
  BTBD11          3296     1  1104  1104 100.0%    12  ++  107712718 108051492 338775   

Gene order in Taeniopygia guttata (finch): 
  RASGRF2         1679     1  1237  1237  95.2%     Z  +-   72041141  72156983 115843
  MSH3            1131   200  1101  1137  82.0%     Z  +-   72194865  72285577  90713
  DHFR_            319     1   187   190  90.5%     Z  ++   72298623  72310674  12052
  ANKRD34B         258    10   183   514  74.8%     Z  ++   72330255  72330776    522
  RASGRF2          739   541   896  1237  85.7%     Z  ++   72356949  72372598  15650

  BTBD11_xenTro   1894   108   997   997  90.1%    1A  +-   53269530  53431966 162437
  CRY1_xenTro     1260    46   574   577  95.3%    1A  +-   53490624  53504662  14039
  MTERFD3_xenTro   146   132   368   369  63.1%    1A  ++   53539437  53540147    711
  C12orf23_homSa   225    20   116   116  88.7%    1A  +-   53545152  53545442    291
  RIC8B_xenTro     693     1   404   414  83.1%    1A  +-   53588230  53608076  19847
  RFX4_xenTro     1422    29   691   691  91.2%    1A  +-   53628722  53681344  52623

  BTBD11_xenTro    164   445   840   997  74.9%     5  +-    6143735   6154710  10976
  SIRT3_xenTro     198   131   311   401  73.9%     5  ++    7212746   7213932   1187
  RIC8B_xenTro     207    15   380   414  72.0%     5  +-    7219279   7223031   3753

  CRY1_xenTro      793    53   488   577  88.9%     5  +-   22665497  22682488  16992
  CHST2_xenTro     700     1   406   407  80.0%     5  ++   22765821  22767074   1254
  C12orf23_homSa   138    25   116   116  75.0%     5  +-   22774918  22775193    276

Gene order in Anolis carolinensis (lizard):
  FAM151B          154    72   171   276  81.4%     2  ++   17132292  17134230   1939
  DHFR_galGal      218     1   153   190  87.9%     2  +-   17184692  17203894  19203
  MSH3            1054   202  1094  1137  82.3%     2  ++   17214220  17270142  55923
  RASGRF2         2613    98  1237  1237  93.1%     2  ++   17343088  17434824  91737

  RFX4_xenTro     1453     1   691   691  91.0%     5  ++   19285834  19332634  46801
  RIC8B_xenTro     610     2   380   414  80.2%     5  ++   19352614  19369230  16617
  C12orf23_homSa   210    25   116   116  88.1%     5  ++   19412228  19412503    276
  MTERFD3_xenTro    77   133   320   369  57.6%     5  +-   19415576  19416139    564
  CRY1_xenTro     1409     1   575   577  96.3%     5  ++   19436254  19471304  35051
  BTBD11_xenTro   1677     1   997   997  89.7%     5  ++   19529954  19797513 267560 

Gene order in Xenopus tropicalis (frog): 
  FAM151B_xenTro   729     1   257   260  99.4%  GL172749  +-      48220     68819  20600
  ANKRD34B_xenTro  1457    1   501   501  99.8%  GL172749  ++      37857     39410   1554

  DHFR_xenTro      562     1   189   189 100.0%  GL173611  +-      10223     18119   7897
  MSH3_xenTro     2477     1   847   847  99.7%  GL173611  ++      25550     81243  55694
  RASGRF2_xenTro  3396     1  1195  1195  99.8%  GL173611  ++     107705    231228 123524

  RFX4_xenTro     1766    70   691   691  99.7%  GL174569  +-       5720     25164  19445

  RIC8B_xenTro    1175     1   401   414 100.0%  GL173571  +-     232468    255175  22708
  SIRT3_xenTro    1167    10   401   401 100.0%  GL173571  ++     223930    227352   3423
  C12orf23_xenTro   66     1    22   114 100.0%  GL173571  +-     191964    192029     66
  CHST1_xenTro    1171     1   405   405  99.7%  GL173571  ++     175980    184029   8050
  MTERFD3_xenTro  1055    12   369   369  99.2%  GL173571  +-      69636     70712   1077
  DHFRL2_xenTro    254    35   197   384  99.3%  GL173571  ++       7955     10720   2766

  CRY1_xenTro     1627     1   576   577 100.0%  GL173033  +-       9078     44385  35308
  BTBD11_xenTro   2800     1   997   997  99.8%  GL173033  ++     181463    298215 116753

Gene order in Danio rerio (zebrafish): 
  ANKRD34B_danRer  960     1   453   453  98.7%    10  +-    3058846   3060435   1590

  FAM151B_danRer   504     1   169   169 100.0%     5  +-   27809804  27813417   3614
  ANKRD34B_danRer  284     1   162   453  81.6%     5  ++   27800655  27803336   2682
  DHFR_danRer      553     1   190   190  99.0%     5  ++   27782945  27789956   7012

  MSH3_danRer     1381     1   472   472 100.0%     5  ++   52945585  53286850 341266
  RASGRF2_danRer  3257     1  1191  1191  99.3%     5  ++   53315572  53434601 119030
  
  RFX4_danRer     2036     1   715   715  99.7%    18  +-   14920893  14956527  35635
  RIC8B_danRer     947     1   363   363  99.1%    18  +-   14895538  14907434  11897
  SIRT3_xenTro     272   128   356   401  74.6%    18  ++   14871213  14875920   4708
  C12orf23_danRer  328     1   112   116  99.2%    18  +-   14849452  14851702   2251
  CHST1_xenTro     259    57   330   405  67.8%    18  ++   14773066  14773905    840
  MTERFD3_danRer   375    28   360   363  70.3%    18  +-   14735350  14736351   1002
  DHFRL2_danRer    546     1   187   187  99.3%    18  ++   14718538  14725012   6475
  CRY1_danRer     1192     1   498   520  90.2%    18  ++   14692957  14710707  17751
  BTBD11_danRer   2081     7  1008  1008  92.2%    18  +-   14542238  14687242 145005