>PAT1
MAFASSSMAALSRPLAAVSSGLGSALSRASQLLTSGSLSSSPTASHSSTRRFISDGTVGSKGRPDSLFLSDISMSGHRAPLLLG
WLNYLERHLPHVGFFEPIGGRALAGSELSVDRHVELMYRVFNMKGDATRMTGISDTEAAQLIASGKQSEVLDRIYAAYMA
YKAGGELDLCLVEGPGPLMGGTELDAQIAAALNAPVLMAMSGRPNATANDYYNKAMVKRQVFADHKVDVLGVVINGLP
REHHAILSSQLRDKLERAGLPFAGALPEDPVLSSVRLDEVRTALGATQLYGETWLGDVEFDEVVVGSQRLEELLETLVERPM
GRPLVVTSADRLDIVLGLLAAQLSVRGPSVAGVLLTQAGASRITRSYAKSAVDNIFAGLSNNTGASGGGPDGAAAANGSAQ
GSLYRGALLPVLSTDKHLAEALAVIGRMDASILPTSIRKVTQCKMLFDKYIDANAVVTGLQKSRPTRVTPKMFQHTMKAMC
RASPQHIVLPESVDKRVLAAAADVTARGLARVTLLGDPTTVQAEAKKLGLDLSGCHIHNPNSSDRFDKYVDMLVEARKKKG
MTREAAADTLHGDINFFGTMMVAAGDADGMVSGAIHTTASTIRPALQMLKNPASSLVSSIFFMCLPDRVLVYGDCAVNV
SPSAADLAAIATTSADTAAAFGIEPRVAMLSYSTLGSGAGPDVQKVTEAVALVKQQRQDIKVEGPIQYDAAIDPAVAAVKV
KGGSEVAGRATVFVFPDLNTGNNTYKAVQQSTGAIAMGPVMQGLLKPVNDLSRGCTVPDIVNTICVTSIQAMQFKQRTQ
AAVAAAAAPK
>PAT2
MSLNSSTMSRRQAVAGAPAVAPFRHAGLFPRVRLCANRRVARVAPKAAGNGNIAQGEQGFDTLFLSDISLVGQRTPLLLG
FFNYFERHLPHVGFFEPIAAEALASSELRIDRHVELVYKVFNLKGDVRAMTGVQDAEAARMIANGQHSELLDKIYSQYASYK
EGQDLVLVEGPGPLMGGTELDAQIAAALNAPVLMTMTGQPNATVADYYNRAMVKRQVFLDHHVEVLGLVMNGLPRQS
HAILSGQLRDKFAAAGLPFAGAIPTDIMLRNVRLDEVQTAMGAQRLYGDSLLTDVEFDDVVVASQRLEELLEILAERPMGR
PLVVTSADRLDIVLGLLAAQLSVSGPGVAGILLTQAGSARSGRNYARDTIDRIFAGLSSSGLYKGSLLPVLVTDMPLRDAIRKL
DNLDAAILPSSTRKISQCKRLFEQYVDANAVVARLQNMVRPNRMTPKMFMHTLKSMCNATPQHIVLPESEDKRVLAAAA
DVVQRGLAKITLLGDPTTILAEAAKLGLDLSGCNIHNPNTSDRFDKYVDMLVEARKKKGMTREVAADTLHGDVNFFATMM
IVAGDADGMVSGAVHTTASTVRPALQVLKSPDTPLVSSVFIMCLPDRVVVYGDCAVNVNPSAADLAQIAITSNDTAAAFGI
EPRVAMLSYSTLGSGSGPDVQKVSEAVAIVKQRRPDIKVEGPIQYDAAIDPKVAAVKVQGLSEVAGKATVFIFPDLNTGNNT
YKAVQQSTGAIAMGPVMQGLLRPVNDLSRGCTVPDIINTICVTSIQASRMSSAARAAAAKAAVAAV
PAT1 MAFASSSMAALSRPLAAVSSGLGSALSRASQLLTSGSLSSSPTASHSSTRRFISDGTVGS 60
PAT2 MSLNSSTMSRRQ----AVAGAPAVAPFRHAGLFPRVRLCANRRVARVAPKAAGNGNIAQG 56
*:: **:*: . **:.. . * * : *:. *.:. .:: :.: ... . .
PAT1 KGRPDSLFLSDISMSGHRAPLLLGWLNYLERHLPHVGFFEPIGGRALAGSELSVDRHVEL 120
PAT2 EQGFDTLFLSDISLVGQRTPLLLGFFNYFERHLPHVGFFEPIAAEALASSELRIDRHVEL 116
: *:*******: *:*:*****::**:*************...***.*** :******
PAT1 MYRVFNMKGDATRMTGISDTEAAQLIASGKQSEVLDRIYAAYMAYKAGGELDLCLVEGPG 180
PAT2 VYKVFNLKGDVRAMTGVQDAEAARMIANGQHSELLDKIYSQYASYKEG--QDLVLVEGPG 174
:*:***:***. ***:.*:***::**.*::**:**:**: * :** * ** ******
PAT1 PLMGGTELDAQIAAALNAPVLMAMSGRPNATANDYYNKAMVKRQVFADHKVDVLGVVING 240
PAT2 PLMGGTELDAQIAAALNAPVLMTMTGQPNATVADYYNRAMVKRQVFLDHHVEVLGLVMNG 234
**********************:*:*:****. ****:******** **:*:***:*:**
PAT1 LPREHHAILSSQLRDKLERAGLPFAGALPEDPVLSSVRLDEVRTALGATQLYGETWLGDV 300
PAT2 LPRQSHAILSGQLRDKFAAAGLPFAGAIPTDIMLRNVRLDEVQTAMGAQRLYGDSLLTDV 294
***: *****.*****: ********:* * :* .******:**:** :***:: * **
PAT1 EFDEVVVGSQRLEELLETLVERPMGRPLVVTSADRLDIVLGLLAAQLSVRGPSVAGVLLT 360
PAT2 EFDDVVVASQRLEELLEILAERPMGRPLVVTSADRLDIVLGLLAAQLSVSGPGVAGILLT 354
***:***.********* *.***************************** **.***:***
PAT1 QAGASRITRSYAKSAVDNIFAGLSNNTGASGGGPDGAAAANGSAQGSLYRGALLPVLSTD 420
PAT2 QAGSARSGRNYARDTIDRIFAGLSSSG--------------------LYKGSLLPVLVTD 394
***::* *.**:.::*.******.. **:*:***** **
PAT1 KHLAEALAVIGRMDASILPTSIRKVTQCKMLFDKYIDANAVVTGLQ-KSRPTRVTPKMFQ 479
PAT2 MPLRDAIRKLDNLDAAILPSSTRKISQCKRLFEQYVDANAVVARLQNMVRPNRMTPKMFM 454
* :*: :..:**:***:* **::*** **::*:******: ** **.*:*****
PAT1 HTMKAMCRASPQHIVLPESVDKRVLAAAADVTARGLARVTLLGDPTTVQAEAKKLGLDLS 539
PAT2 HTLKSMCNATPQHIVLPESEDKRVLAAAADVVQRGLAKITLLGDPTTILAEAAKLGLDLS 514
**:*:**.*:********* ***********. ****::********: *** *******
PAT1 GCHIHNPNSSDRFDKYVDMLVEARKKKGMTREAAADTLHGDINFFGTMMVAAGDADGMVS 599
PAT2 GCNIHNPNTSDRFDKYVDMLVEARKKKGMTREVAADTLHGDVNFFATMMIVAGDADGMVS 574
**:*****:***********************.********:***.***:.*********
PAT1 GAIHTTASTIRPALQMLKNPASSLVSSIFFMCLPDRVLVYGDCAVNVSPSAADLAAIATT 659
PAT2 GAVHTTASTVRPALQVLKSPDTPLVSSVFIMCLPDRVVVYGDCAVNVNPSAADLAQIAIT 634
**:******:*****:**.* :.****:*:*******:*********.******* ** *
PAT1 SADTAAAFGIEPRVAMLSYSTLGSGAGPDVQKVTEAVALVKQQRQDIKVEGPIQYDAAID 719
PAT2 SNDTAAAFGIEPRVAMLSYSTLGSGSGPDVQKVSEAVAIVKQRRPDIKVEGPIQYDAAID 694
* ***********************:*******:****:***:* ***************
PAT1 PAVAAVKVKGGSEVAGRATVFVFPDLNTGNNTYKAVQQSTGAIAMGPVMQGLLKPVNDLS 779
PAT2 PKVAAVKVQGLSEVAGKATVFIFPDLNTGNNTYKAVQQSTGAIAMGPVMQGLLRPVNDLS 754
* ******:* *****:****:*******************************:******
PAT1 RGCTVPDIVNTICVTSIQAMQFKQRTQAAVAAAAAPK- 816
PAT2 RGCTVPDIINTICVTSIQASRMSSAARAAAAKAAVAAV 792
********:********** ::.. ::**.* **..
>ACK2
MKPRKWELKVPAQDHVGAMKGILDFIKENVSATFASEVVAVGHRIVHGLDLSQPVLLTPSSTATIKQAAVLAPLHNPPGLQ
GIEAATEVFSGVPQVAVFDTAFHQTMPPHAYMYGLPYELYEKHKIRRYGFHGTSHKYLVEQAAAMLGKPVSDTNVITCHL
GNGSSVAAVRGGQCVDTSMGLTPLEGLLMGTRCGDMDPAVVLHIQNQCGLNVKETDTLLNKKSGLLGLTGSNDLRAVIE
GAGKGELRSQLGLDMFTYRVRKYIGAYTAALDGKVDAVVFSAGIGENSPFIRKLIAEPLHAMGMQLDHAANDTAVGRAGV
ISTPGSRVQLLVIPTDEELSIAEQTLQVAKGALGAAAA
>ACK1
MLAGKVPVGSAVPPRNTAPSGKGSKVLVVNAGSSSIKFKIFTAETLLPGAAGLFDRIGDPANCVLKASVPDDGGKPKKFEKKI
PAKDHLTAMQDIMSFIDENVSSTFASEVCAVGHRIVHGLDMSQPKLLTPDVIRCIQDASMFAPLHNPPHLQGIYASQKVFG
PDVPQVAVFDTAFHQTMPPESYVYGLPYDMYEKHKIRRYGFHGTSHKYLVDQAAAMLGKKVSETNVITCHLGNGSSVAA
VRGGQCVDTSMGMTPLEGLLMGTRCGDIDPAVVLHIQNQLGLSASETDTLLNKKSGLLGLTGSNDLRTVIEGAGKGEPRS
QLGLDMFVRRVRKYIGAYMAMLDGQVDAVVFSAGIGENSALVRGLICDRMQALGIAVDKDANAAAVGGKQADISGPGA
RIRALVIPTDEELSIAQQTLQVVRGAALAA
ACK2 ------------------------------------------------------------
ACK1 MLAGKVPVGSAVPPRNTAPSGKGSKVLVVNAGSSSIKFKIFTAETLLPGAAGLFDRIGDP 60
ACK2 -------------MKPRKWELKVPAQDHVGAMKGILDFIKENVSATFASEVVAVGHRIVH 47
ACK1 ANCVLKASVPDDGGKPKKFEKKIPAKDHLTAMQDIMSFIDENVSSTFASEVCAVGHRIVH 120
**:*:* *:**:**: **:.*:.**.****:****** ********
ACK2 GLDLSQPVLLTPSSTATIKQAAVLAPLHNPPGLQGIEAATEVFS-GVPQVAVFDTAFHQT 106
ACK1 GLDMSQPKLLTPDVIRCIQDASMFAPLHNPPHLQGIYASQKVFGPDVPQVAVFDTAFHQT 180
***:*** ****. *::*:::******* **** *: :**. .**************
ACK2 MPPHAYMYGLPYELYEKHKIRRYGFHGTSHKYLVEQAAAMLGKPVSDTNVITCHLGNGSS 166
ACK1 MPPESYVYGLPYDMYEKHKIRRYGFHGTSHKYLVDQAAAMLGKKVSETNVITCHLGNGSS 240
***.:*:*****::********************:******** **:*************
ACK2 VAAVRGGQCVDTSMGLTPLEGLLMGTRCGDMDPAVVLHIQNQCGLNVKETDTLLNKKSGL 226
ACK1 VAAVRGGQCVDTSMGMTPLEGLLMGTRCGDIDPAVVLHIQNQLGLSASETDTLLNKKSGL 300
***************:**************:*********** **...************
ACK2 LGLTGSNDLRAVIEGAGKGELRSQLGLDMFTYRVRKYIGAYTAALDGKVDAVVFSAGIGE 286
ACK1 LGLTGSNDLRTVIEGAGKGEPRSQLGLDMFVRRVRKYIGAYMAMLDGQVDAVVFSAGIGE 360
**********:********* *********. ********* * ***:************
ACK2 NSPFIRKLIAEPLHAMGMQLDHAANDTAVG-RAGVISTPGSRVQLLVIPTDEELSIAEQT 345
ACK1 NSALVRGLICDRMQALGIAVDKDANAAAVGGKQADISGPGARIRALVIPTDEELSIAQQT 420
**.::* **.: ::*:*: :*: ** :*** : . ** **:*:: ************:**
ACK2 LQVAKGALGAAAA 358
ACK1 LQVVRGAALAA-- 431
***.:** **
Supplemental Figure S2: Protein sequence alignment of Chlamydomonas reinhardtii PAT and ACK isoforms.
Sequences are from PAT1 (JGI #11226; Augustus #517066; Predicted MW 85.68 KDa), PAT2 (JGI #191051;
Augustus #526017; Predicted MW 84.63 KDa), ACK2 (JGI #128476; Augustus #517287; Predicted MW 37.94
KDa) and ACK1 (JGI #129982; Augustus #526019; Predicted MW 45.53 KDa). PAT2 and ACK1 are the
chloroplast-localized isoforms and are highlighted in green. ACK1 antibodies were raised against the full-
length recombinant protein, whereas PAT2 antibodies were generated against the synthesized peptide
RSGRNYARDTIDRIF (highlighted in yellow), conjugated to KLH (Keyhole Limpet Hemocyanin) carrier protein
via a cysteine that was added to the N-terminus of the peptide.
