U.S. patent application number 15/194925 was filed with the patent office on 2016-10-20 for synthetic genes. The applicant listed for this patent is Dow AgroSciences LLC. Invention is credited to Ignacio M. Larrinua, Donald Merlo, Avutu Sambi Reddy, Arvind Kumar ThirumalaiswamySekhar, Aaron T. Woosley.
Application Number | 20160304900 15/194925 |
Document ID | / |
Family ID | 47007465 |
Filed Date | 2016-10-20 |
United States Patent Application | 20160304900 |
Kind Code | A1 |
Larrinua; Ignacio M. ; et al. | October 20, 2016 |
The invention provides synthetic nucleic acid sequences encoding proteins of interest that are particularly adapted to express well in plants. The claimed synthetic sequences utilize plant-optimized codons roughly in the same frequency at which they are utilized, on average, in genes naturally occurring in the plant species. The invention further includes synthetic DNA sequence for herbicide tolerance, water and/or heat stress tolerance, healthy oil modifications and for transformation marker genes and selectable marker genes are used. DNA construct and transgenic plants containing the synthetic sequences are taught as are methods and compositions for using the plants in agriculture.
Inventors: | Larrinua; Ignacio M.; (Indianapolis, IN) ; Merlo; Donald; (Carmel, IN) ; Reddy; Avutu Sambi; (Carmel, IN) ; ThirumalaiswamySekhar; Arvind Kumar; (Zionsville, IN) ; Woosley; Aaron T.; (Fishers, IN) | ||||||||||
Applicant: |
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Family ID: | 47007465 | ||||||||||
Appl. No.: | 15/194925 | ||||||||||
Filed: | June 28, 2016 |
Application Number | Filing Date | Patent Number | ||
---|---|---|---|---|
13447836 | Apr 16, 2012 | 9427003 | ||
15194925 | ||||
61475921 | Apr 15, 2011 | |||
Current U.S. Class: | 1/1 |
Current CPC Class: | C07K 14/195 20130101; A23L 7/198 20160801; C11B 1/00 20130101; A23L 7/10 20160801; A23L 19/10 20160801; C07K 14/38 20130101; C07K 14/43595 20130101; C07K 14/415 20130101; C07K 14/37 20130101; C07K 14/325 20130101; C12Y 111/01015 20130101; Y02A 40/162 20180101; A23L 11/05 20160801; C12N 9/0069 20130101; C12N 9/0083 20130101; C12N 9/0065 20130101; C12N 15/8286 20130101; A23L 11/00 20160801; A23D 9/00 20130101; C12Y 113/00 20130101; Y02A 40/146 20180101 |
International Class: | C12N 15/82 20060101 C12N015/82; C07K 14/325 20060101 C07K014/325; C07K 14/37 20060101 C07K014/37; C07K 14/38 20060101 C07K014/38; C07K 14/415 20060101 C07K014/415; C07K 14/435 20060101 C07K014/435; C12N 9/02 20060101 C12N009/02; C07K 14/195 20060101 C07K014/195 |
Sequence CWU 1
1
7211818DNABacillus thuringiensisCDS(1)..(1818)Native DNA sequence
encoding Bacillus thuringiensis Cry1Fa core toxin 1atg gag aat aat
att caa aat caa tgc gta cct tac aat tgt tta aat 48Met Glu Asn Asn
Ile Gln Asn Gln Cys Val Pro Tyr Asn Cys Leu Asn 1 5 10 15 aat cct
gaa gta gaa ata tta aat gaa gaa aga agt act ggc aga tta 96Asn Pro
Glu Val Glu Ile Leu Asn Glu Glu Arg Ser Thr Gly Arg Leu 20 25 30
ccg tta gat ata tcc tta tcg ctt aca cgt ttc ctt ttg agt gaa ttt
144Pro Leu Asp Ile Ser Leu Ser Leu Thr Arg Phe Leu Leu Ser Glu Phe
35 40 45 gtt cca ggt gtg gga gtt gcg ttt gga tta ttt gat tta ata
tgg ggt 192Val Pro Gly Val Gly Val Ala Phe Gly Leu Phe Asp Leu Ile
Trp Gly 50 55 60 ttt ata act cct tct gat tgg agc tta ttt ctt tta
cag att gaa caa 240Phe Ile Thr Pro Ser Asp Trp Ser Leu Phe Leu Leu
Gln Ile Glu Gln 65 70 75 80 ttg att gag caa aga ata gaa aca ttg gaa
agg aac cgg gca att act 288Leu Ile Glu Gln Arg Ile Glu Thr Leu Glu
Arg Asn Arg Ala Ile Thr 85 90 95 aca tta cga ggg tta gca gat agc
tat gaa att tat att gaa gca cta 336Thr Leu Arg Gly Leu Ala Asp Ser
Tyr Glu Ile Tyr Ile Glu Ala Leu 100 105 110 aga gag tgg gaa gca aat
cct aat aat gca caa tta agg gaa gat gtg 384Arg Glu Trp Glu Ala Asn
Pro Asn Asn Ala Gln Leu Arg Glu Asp Val 115 120 125 cgt att cga ttt
gct aat aca gac gac gct tta ata aca gca ata aat 432Arg Ile Arg Phe
Ala Asn Thr Asp Asp Ala Leu Ile Thr Ala Ile Asn 130 135 140 aat ttt
aca ctt aca agt ttt gaa atc cct ctt tta tcg gtc tat gtt 480Asn Phe
Thr Leu Thr Ser Phe Glu Ile Pro Leu Leu Ser Val Tyr Val 145 150 155
160 caa gcg gcg aat tta cat tta tca cta tta aga gac gct gta tcg ttt
528Gln Ala Ala Asn Leu His Leu Ser Leu Leu Arg Asp Ala Val Ser Phe
165 170 175 ggg cag ggt tgg gga ctg gat ata gct act gtt aat aat cat
tat aat 576Gly Gln Gly Trp Gly Leu Asp Ile Ala Thr Val Asn Asn His
Tyr Asn 180 185 190 aga tta ata aat ctt att cat aga tat acg aaa cat
tgt ttg gac aca 624Arg Leu Ile Asn Leu Ile His Arg Tyr Thr Lys His
Cys Leu Asp Thr 195 200 205 tac aat caa gga tta gaa aac tta aga ggt
act aat act cga caa tgg 672Tyr Asn Gln Gly Leu Glu Asn Leu Arg Gly
Thr Asn Thr Arg Gln Trp 210 215 220 gca aga ttc aat cag ttt agg aga
gat tta aca ctt act gta tta gat 720Ala Arg Phe Asn Gln Phe Arg Arg
Asp Leu Thr Leu Thr Val Leu Asp 225 230 235 240 atc gtt gct ctt ttt
ccg aac tac gat gtt aga aca tat cca att caa 768Ile Val Ala Leu Phe
Pro Asn Tyr Asp Val Arg Thr Tyr Pro Ile Gln 245 250 255 acg tca tcc
caa tta aca agg gaa att tat aca agt tca gta att gag 816Thr Ser Ser
Gln Leu Thr Arg Glu Ile Tyr Thr Ser Ser Val Ile Glu 260 265 270 gat
tct cca gtt tct gct aat ata cct aat ggt ttt aat agg gcg gaa 864Asp
Ser Pro Val Ser Ala Asn Ile Pro Asn Gly Phe Asn Arg Ala Glu 275 280
285 ttt gga gtt aga ccg ccc cat ctt atg gac ttt atg aat tct ttg ttt
912Phe Gly Val Arg Pro Pro His Leu Met Asp Phe Met Asn Ser Leu Phe
290 295 300 gta act gca gag act gtt aga agt caa act gtg tgg gga gga
cac tta 960Val Thr Ala Glu Thr Val Arg Ser Gln Thr Val Trp Gly Gly
His Leu 305 310 315 320 gtt agt tca cga aat acg gct ggt aac cgt ata
aat ttc cct agt tac 1008Val Ser Ser Arg Asn Thr Ala Gly Asn Arg Ile
Asn Phe Pro Ser Tyr 325 330 335 ggg gtc ttc aat cct ggt ggc gcc att
tgg att gca gat gag gat cca 1056Gly Val Phe Asn Pro Gly Gly Ala Ile
Trp Ile Ala Asp Glu Asp Pro 340 345 350 cgt cct ttt tat cgg aca tta
tca gat cct gtt ttt gtc cga gga gga 1104Arg Pro Phe Tyr Arg Thr Leu
Ser Asp Pro Val Phe Val Arg Gly Gly 355 360 365 ttt ggg aat cct cat
tat gta ctg ggg ctt agg gga gta gca ttt caa 1152Phe Gly Asn Pro His
Tyr Val Leu Gly Leu Arg Gly Val Ala Phe Gln 370 375 380 caa act ggt
acg aac cac acc cga aca ttt aga aat agt ggg acc ata 1200Gln Thr Gly
Thr Asn His Thr Arg Thr Phe Arg Asn Ser Gly Thr Ile 385 390 395 400
gat tct cta gat gaa atc cca cct cag gat aat agt ggg gca cct tgg
1248Asp Ser Leu Asp Glu Ile Pro Pro Gln Asp Asn Ser Gly Ala Pro Trp
405 410 415 aat gat tat agt cat gta tta aat cat gtt aca ttt gta cga
tgg cca 1296Asn Asp Tyr Ser His Val Leu Asn His Val Thr Phe Val Arg
Trp Pro 420 425 430 ggt gag att tca gga agt gat tca tgg aga gct cca
atg ttt tct tgg 1344Gly Glu Ile Ser Gly Ser Asp Ser Trp Arg Ala Pro
Met Phe Ser Trp 435 440 445 acg cac cgt agt gca acc cct aca aat aca
att gat ccg gag agg att 1392Thr His Arg Ser Ala Thr Pro Thr Asn Thr
Ile Asp Pro Glu Arg Ile 450 455 460 act caa ata cca ttg gta aaa gca
cat aca ctt cag tca ggt act act 1440Thr Gln Ile Pro Leu Val Lys Ala
His Thr Leu Gln Ser Gly Thr Thr 465 470 475 480 gtt gta aga ggg ccc
ggg ttt acg gga gga gat att ctt cga cga aca 1488Val Val Arg Gly Pro
Gly Phe Thr Gly Gly Asp Ile Leu Arg Arg Thr 485 490 495 agt gga gga
cca ttt gct tat act att gtt aat ata aat ggg caa tta 1536Ser Gly Gly
Pro Phe Ala Tyr Thr Ile Val Asn Ile Asn Gly Gln Leu 500 505 510 ccc
caa agg tat cgt gca aga ata cgc tat gcc tct act aca aat cta 1584Pro
Gln Arg Tyr Arg Ala Arg Ile Arg Tyr Ala Ser Thr Thr Asn Leu 515 520
525 aga att tac gta acg gtt gca ggt gaa cgg att ttt gct ggt caa ttt
1632Arg Ile Tyr Val Thr Val Ala Gly Glu Arg Ile Phe Ala Gly Gln Phe
530 535 540 aac aaa aca atg gat acc ggt gac cca tta aca ttc caa tct
ttt agt 1680Asn Lys Thr Met Asp Thr Gly Asp Pro Leu Thr Phe Gln Ser
Phe Ser 545 550 555 560 tac gca act att aat aca gct ttt aca ttc cca
atg agc cag agt agt 1728Tyr Ala Thr Ile Asn Thr Ala Phe Thr Phe Pro
Met Ser Gln Ser Ser 565 570 575 ttc aca gta ggt gct gat act ttt agt
tca ggg aat gaa gtt tat ata 1776Phe Thr Val Gly Ala Asp Thr Phe Ser
Ser Gly Asn Glu Val Tyr Ile 580 585 590 gac aga ttt gaa ttg att cca
gtt act gca aca ttt gaa tag 1818Asp Arg Phe Glu Leu Ile Pro Val Thr
Ala Thr Phe Glu 595 600 605 2605PRTBacillus thuringiensis 2Met Glu
Asn Asn Ile Gln Asn Gln Cys Val Pro Tyr Asn Cys Leu Asn 1 5 10 15
Asn Pro Glu Val Glu Ile Leu Asn Glu Glu Arg Ser Thr Gly Arg Leu 20
25 30 Pro Leu Asp Ile Ser Leu Ser Leu Thr Arg Phe Leu Leu Ser Glu
Phe 35 40 45 Val Pro Gly Val Gly Val Ala Phe Gly Leu Phe Asp Leu
Ile Trp Gly 50 55 60 Phe Ile Thr Pro Ser Asp Trp Ser Leu Phe Leu
Leu Gln Ile Glu Gln 65 70 75 80 Leu Ile Glu Gln Arg Ile Glu Thr Leu
Glu Arg Asn Arg Ala Ile Thr 85 90 95 Thr Leu Arg Gly Leu Ala Asp
Ser Tyr Glu Ile Tyr Ile Glu Ala Leu 100 105 110 Arg Glu Trp Glu Ala
Asn Pro Asn Asn Ala Gln Leu Arg Glu Asp Val 115 120 125 Arg Ile Arg
Phe Ala Asn Thr Asp Asp Ala Leu Ile Thr Ala Ile Asn 130 135 140 Asn
Phe Thr Leu Thr Ser Phe Glu Ile Pro Leu Leu Ser Val Tyr Val 145 150
155 160 Gln Ala Ala Asn Leu His Leu Ser Leu Leu Arg Asp Ala Val Ser
Phe 165 170 175 Gly Gln Gly Trp Gly Leu Asp Ile Ala Thr Val Asn Asn
His Tyr Asn 180 185 190 Arg Leu Ile Asn Leu Ile His Arg Tyr Thr Lys
His Cys Leu Asp Thr 195 200 205 Tyr Asn Gln Gly Leu Glu Asn Leu Arg
Gly Thr Asn Thr Arg Gln Trp 210 215 220 Ala Arg Phe Asn Gln Phe Arg
Arg Asp Leu Thr Leu Thr Val Leu Asp 225 230 235 240 Ile Val Ala Leu
Phe Pro Asn Tyr Asp Val Arg Thr Tyr Pro Ile Gln 245 250 255 Thr Ser
Ser Gln Leu Thr Arg Glu Ile Tyr Thr Ser Ser Val Ile Glu 260 265 270
Asp Ser Pro Val Ser Ala Asn Ile Pro Asn Gly Phe Asn Arg Ala Glu 275
280 285 Phe Gly Val Arg Pro Pro His Leu Met Asp Phe Met Asn Ser Leu
Phe 290 295 300 Val Thr Ala Glu Thr Val Arg Ser Gln Thr Val Trp Gly
Gly His Leu 305 310 315 320 Val Ser Ser Arg Asn Thr Ala Gly Asn Arg
Ile Asn Phe Pro Ser Tyr 325 330 335 Gly Val Phe Asn Pro Gly Gly Ala
Ile Trp Ile Ala Asp Glu Asp Pro 340 345 350 Arg Pro Phe Tyr Arg Thr
Leu Ser Asp Pro Val Phe Val Arg Gly Gly 355 360 365 Phe Gly Asn Pro
His Tyr Val Leu Gly Leu Arg Gly Val Ala Phe Gln 370 375 380 Gln Thr
Gly Thr Asn His Thr Arg Thr Phe Arg Asn Ser Gly Thr Ile 385 390 395
400 Asp Ser Leu Asp Glu Ile Pro Pro Gln Asp Asn Ser Gly Ala Pro Trp
405 410 415 Asn Asp Tyr Ser His Val Leu Asn His Val Thr Phe Val Arg
Trp Pro 420 425 430 Gly Glu Ile Ser Gly Ser Asp Ser Trp Arg Ala Pro
Met Phe Ser Trp 435 440 445 Thr His Arg Ser Ala Thr Pro Thr Asn Thr
Ile Asp Pro Glu Arg Ile 450 455 460 Thr Gln Ile Pro Leu Val Lys Ala
His Thr Leu Gln Ser Gly Thr Thr 465 470 475 480 Val Val Arg Gly Pro
Gly Phe Thr Gly Gly Asp Ile Leu Arg Arg Thr 485 490 495 Ser Gly Gly
Pro Phe Ala Tyr Thr Ile Val Asn Ile Asn Gly Gln Leu 500 505 510 Pro
Gln Arg Tyr Arg Ala Arg Ile Arg Tyr Ala Ser Thr Thr Asn Leu 515 520
525 Arg Ile Tyr Val Thr Val Ala Gly Glu Arg Ile Phe Ala Gly Gln Phe
530 535 540 Asn Lys Thr Met Asp Thr Gly Asp Pro Leu Thr Phe Gln Ser
Phe Ser 545 550 555 560 Tyr Ala Thr Ile Asn Thr Ala Phe Thr Phe Pro
Met Ser Gln Ser Ser 565 570 575 Phe Thr Val Gly Ala Asp Thr Phe Ser
Ser Gly Asn Glu Val Tyr Ile 580 585 590 Asp Arg Phe Glu Leu Ile Pro
Val Thr Ala Thr Phe Glu 595 600 605 31818DNAArtificial
SequenceSynthetic DNA sequence encoding Bacillus thuringiensis
Cry1Fa core toxin using codons optimized for maize and Table 1
sequences are maintained 3atg gag aat aat atc cag aat caa tgc gtg
cct tac aat tgt tta aat 48Met Glu Asn Asn Ile Gln Asn Gln Cys Val
Pro Tyr Asn Cys Leu Asn 1 5 10 15 aat ccc gag gtg gag ata tta aac
gag gag aga tcc act ggc aga ctg 96Asn Pro Glu Val Glu Ile Leu Asn
Glu Glu Arg Ser Thr Gly Arg Leu 20 25 30 cca ctc gat ata tcc ttg
tcc ctt acc cgt ttc ctt ttg agc gaa ttt 144Pro Leu Asp Ile Ser Leu
Ser Leu Thr Arg Phe Leu Leu Ser Glu Phe 35 40 45 gtt cct ggt gtg
gga gtg gct ttc gga tta ttt gat ctg ata tgg ggt 192Val Pro Gly Val
Gly Val Ala Phe Gly Leu Phe Asp Leu Ile Trp Gly 50 55 60 ttt atc
act cct tct gat tgg agc tta ttt ctt ctc cag att gag caa 240Phe Ile
Thr Pro Ser Asp Trp Ser Leu Phe Leu Leu Gln Ile Glu Gln 65 70 75 80
ttg att gag cag aga ata gaa acc ttg gaa agg aac cgt gca atc acg
288Leu Ile Glu Gln Arg Ile Glu Thr Leu Glu Arg Asn Arg Ala Ile Thr
85 90 95 acc ttg cgc ggt ctc gcc gat agc tat gaa att tat att gaa
gca ctg 336Thr Leu Arg Gly Leu Ala Asp Ser Tyr Glu Ile Tyr Ile Glu
Ala Leu 100 105 110 agg gag tgg gag gcc aac cct aat aat gct caa tta
agg gaa gat gtg 384Arg Glu Trp Glu Ala Asn Pro Asn Asn Ala Gln Leu
Arg Glu Asp Val 115 120 125 cgt att cgt ttt gct aat aca gac gac gct
tta ata aca gca ata aat 432Arg Ile Arg Phe Ala Asn Thr Asp Asp Ala
Leu Ile Thr Ala Ile Asn 130 135 140 aat ttc aca ctt aca tcc ttt gaa
atc ccg ctt tta tca gtg tac gtt 480Asn Phe Thr Leu Thr Ser Phe Glu
Ile Pro Leu Leu Ser Val Tyr Val 145 150 155 160 caa gcc gcc aat ctc
cat tta tca ctt ctg agg gac gct gtc tcc ttt 528Gln Ala Ala Asn Leu
His Leu Ser Leu Leu Arg Asp Ala Val Ser Phe 165 170 175 ggg caa ggt
tgg gga ctg gat atc gct act gtt aat aat cac tac aat 576Gly Gln Gly
Trp Gly Leu Asp Ile Ala Thr Val Asn Asn His Tyr Asn 180 185 190 aga
tta ata aac ctg att cat aga tat acg aag cat tgt ttg gac aca 624Arg
Leu Ile Asn Leu Ile His Arg Tyr Thr Lys His Cys Leu Asp Thr 195 200
205 tac aat caa gga ctg gag aac ctt agg gga act aac act agg cag tgg
672Tyr Asn Gln Gly Leu Glu Asn Leu Arg Gly Thr Asn Thr Arg Gln Trp
210 215 220 gca agg ttc aac cag ttc aga cgt gat ctc aca ctt act gtg
ctg gat 720Ala Arg Phe Asn Gln Phe Arg Arg Asp Leu Thr Leu Thr Val
Leu Asp 225 230 235 240 atc gtt gct ctc ttt ccg aac tac gat gtt cgc
acc tac cca atc cag 768Ile Val Ala Leu Phe Pro Asn Tyr Asp Val Arg
Thr Tyr Pro Ile Gln 245 250 255 acg tca tcc caa tta aca agg gaa att
tat acc tcc tca gtg att gag 816Thr Ser Ser Gln Leu Thr Arg Glu Ile
Tyr Thr Ser Ser Val Ile Glu 260 265 270 gac tct ccc gtt tct gct aac
ata cct aac ggc ttc aac cgc gcc gag 864Asp Ser Pro Val Ser Ala Asn
Ile Pro Asn Gly Phe Asn Arg Ala Glu 275 280 285 ttc gga gtt aga ccg
ccc cac ctt atg gac ttt atg aat agc ttg ttt 912Phe Gly Val Arg Pro
Pro His Leu Met Asp Phe Met Asn Ser Leu Phe 290 295 300 gtg act gct
gag act gtt aga agc caa act gtg tgg ggc ggc cac ttg 960Val Thr Ala
Glu Thr Val Arg Ser Gln Thr Val Trp Gly Gly His Leu 305 310 315 320
gtc agc tca cgc aac acg gct ggc aac cgt atc aac ttc ccg tct tac
1008Val Ser Ser Arg Asn Thr Ala Gly Asn Arg Ile Asn Phe Pro Ser Tyr
325 330 335 ggg gtc ttt aac cct ggt ggc gcc att tgg att gca gac gag
gac cca 1056Gly Val Phe Asn Pro Gly Gly Ala Ile Trp Ile Ala Asp Glu
Asp Pro 340 345 350 cgt cct ttt tat cgc acc ctg tca gat cct gtt ttt
gtc aga ggc gga 1104Arg Pro Phe Tyr Arg Thr Leu Ser Asp Pro Val Phe
Val Arg Gly Gly
355 360 365 ttt ggg aat cct cat tat gtc ctg ggc ctt agg gga gtg gct
ttc caa 1152Phe Gly Asn Pro His Tyr Val Leu Gly Leu Arg Gly Val Ala
Phe Gln 370 375 380 cag act ggc acc aac cac acc cgt acg ttt cgc aat
agc ggg acc ata 1200Gln Thr Gly Thr Asn His Thr Arg Thr Phe Arg Asn
Ser Gly Thr Ile 385 390 395 400 gat tct ctt gat gaa atc cca cct caa
gat aac agc ggc gca cct tgg 1248Asp Ser Leu Asp Glu Ile Pro Pro Gln
Asp Asn Ser Gly Ala Pro Trp 405 410 415 aac gat tat tcc cac gta tta
aat cac gtt acg ttc gtc cgc tgg ccg 1296Asn Asp Tyr Ser His Val Leu
Asn His Val Thr Phe Val Arg Trp Pro 420 425 430 ggt gag atc agc ggc
agc gat tca tgg aga gca cca atg ttt tct tgg 1344Gly Glu Ile Ser Gly
Ser Asp Ser Trp Arg Ala Pro Met Phe Ser Trp 435 440 445 acg cac cgt
tca gcc acc cct aca aat aca att gac ccg gag agg att 1392Thr His Arg
Ser Ala Thr Pro Thr Asn Thr Ile Asp Pro Glu Arg Ile 450 455 460 act
caa atc cca ttg gtc aaa gca cat aca ctt cag tct ggg acc acc 1440Thr
Gln Ile Pro Leu Val Lys Ala His Thr Leu Gln Ser Gly Thr Thr 465 470
475 480 gtg gtc aga ggg cct ggg ttc acg gga gga gac att ctt agg cgc
aca 1488Val Val Arg Gly Pro Gly Phe Thr Gly Gly Asp Ile Leu Arg Arg
Thr 485 490 495 tcc gga gga ccc ttc gct tat act atc gtt aat ata aat
ggg cag ctc 1536Ser Gly Gly Pro Phe Ala Tyr Thr Ile Val Asn Ile Asn
Gly Gln Leu 500 505 510 ccc cag cgc tat cgt gcc aga atc cgt tac gcc
tct act aca aat ctc 1584Pro Gln Arg Tyr Arg Ala Arg Ile Arg Tyr Ala
Ser Thr Thr Asn Leu 515 520 525 aga atc tac gtg acg gtt gcc ggt gag
cgc att ttt gct ggt cag ttt 1632Arg Ile Tyr Val Thr Val Ala Gly Glu
Arg Ile Phe Ala Gly Gln Phe 530 535 540 aac aag acg atg gat act ggc
gac cca ctg aca ttc caa tct ttc tca 1680Asn Lys Thr Met Asp Thr Gly
Asp Pro Leu Thr Phe Gln Ser Phe Ser 545 550 555 560 tac gca act att
aat aca gct ttc aca ttc cca atg agc cag tca tct 1728Tyr Ala Thr Ile
Asn Thr Ala Phe Thr Phe Pro Met Ser Gln Ser Ser 565 570 575 ttc acc
gtc ggt gct gat acc ttc agc tct ggc aac gaa gtt tat ata 1776Phe Thr
Val Gly Ala Asp Thr Phe Ser Ser Gly Asn Glu Val Tyr Ile 580 585 590
gac aga ttt gag ttg att cca gtt act gca acg ttt gag tga 1818Asp Arg
Phe Glu Leu Ile Pro Val Thr Ala Thr Phe Glu 595 600 605
4605PRTArtificial SequenceSynthetic Construct 4Met Glu Asn Asn Ile
Gln Asn Gln Cys Val Pro Tyr Asn Cys Leu Asn 1 5 10 15 Asn Pro Glu
Val Glu Ile Leu Asn Glu Glu Arg Ser Thr Gly Arg Leu 20 25 30 Pro
Leu Asp Ile Ser Leu Ser Leu Thr Arg Phe Leu Leu Ser Glu Phe 35 40
45 Val Pro Gly Val Gly Val Ala Phe Gly Leu Phe Asp Leu Ile Trp Gly
50 55 60 Phe Ile Thr Pro Ser Asp Trp Ser Leu Phe Leu Leu Gln Ile
Glu Gln 65 70 75 80 Leu Ile Glu Gln Arg Ile Glu Thr Leu Glu Arg Asn
Arg Ala Ile Thr 85 90 95 Thr Leu Arg Gly Leu Ala Asp Ser Tyr Glu
Ile Tyr Ile Glu Ala Leu 100 105 110 Arg Glu Trp Glu Ala Asn Pro Asn
Asn Ala Gln Leu Arg Glu Asp Val 115 120 125 Arg Ile Arg Phe Ala Asn
Thr Asp Asp Ala Leu Ile Thr Ala Ile Asn 130 135 140 Asn Phe Thr Leu
Thr Ser Phe Glu Ile Pro Leu Leu Ser Val Tyr Val 145 150 155 160 Gln
Ala Ala Asn Leu His Leu Ser Leu Leu Arg Asp Ala Val Ser Phe 165 170
175 Gly Gln Gly Trp Gly Leu Asp Ile Ala Thr Val Asn Asn His Tyr Asn
180 185 190 Arg Leu Ile Asn Leu Ile His Arg Tyr Thr Lys His Cys Leu
Asp Thr 195 200 205 Tyr Asn Gln Gly Leu Glu Asn Leu Arg Gly Thr Asn
Thr Arg Gln Trp 210 215 220 Ala Arg Phe Asn Gln Phe Arg Arg Asp Leu
Thr Leu Thr Val Leu Asp 225 230 235 240 Ile Val Ala Leu Phe Pro Asn
Tyr Asp Val Arg Thr Tyr Pro Ile Gln 245 250 255 Thr Ser Ser Gln Leu
Thr Arg Glu Ile Tyr Thr Ser Ser Val Ile Glu 260 265 270 Asp Ser Pro
Val Ser Ala Asn Ile Pro Asn Gly Phe Asn Arg Ala Glu 275 280 285 Phe
Gly Val Arg Pro Pro His Leu Met Asp Phe Met Asn Ser Leu Phe 290 295
300 Val Thr Ala Glu Thr Val Arg Ser Gln Thr Val Trp Gly Gly His Leu
305 310 315 320 Val Ser Ser Arg Asn Thr Ala Gly Asn Arg Ile Asn Phe
Pro Ser Tyr 325 330 335 Gly Val Phe Asn Pro Gly Gly Ala Ile Trp Ile
Ala Asp Glu Asp Pro 340 345 350 Arg Pro Phe Tyr Arg Thr Leu Ser Asp
Pro Val Phe Val Arg Gly Gly 355 360 365 Phe Gly Asn Pro His Tyr Val
Leu Gly Leu Arg Gly Val Ala Phe Gln 370 375 380 Gln Thr Gly Thr Asn
His Thr Arg Thr Phe Arg Asn Ser Gly Thr Ile 385 390 395 400 Asp Ser
Leu Asp Glu Ile Pro Pro Gln Asp Asn Ser Gly Ala Pro Trp 405 410 415
Asn Asp Tyr Ser His Val Leu Asn His Val Thr Phe Val Arg Trp Pro 420
425 430 Gly Glu Ile Ser Gly Ser Asp Ser Trp Arg Ala Pro Met Phe Ser
Trp 435 440 445 Thr His Arg Ser Ala Thr Pro Thr Asn Thr Ile Asp Pro
Glu Arg Ile 450 455 460 Thr Gln Ile Pro Leu Val Lys Ala His Thr Leu
Gln Ser Gly Thr Thr 465 470 475 480 Val Val Arg Gly Pro Gly Phe Thr
Gly Gly Asp Ile Leu Arg Arg Thr 485 490 495 Ser Gly Gly Pro Phe Ala
Tyr Thr Ile Val Asn Ile Asn Gly Gln Leu 500 505 510 Pro Gln Arg Tyr
Arg Ala Arg Ile Arg Tyr Ala Ser Thr Thr Asn Leu 515 520 525 Arg Ile
Tyr Val Thr Val Ala Gly Glu Arg Ile Phe Ala Gly Gln Phe 530 535 540
Asn Lys Thr Met Asp Thr Gly Asp Pro Leu Thr Phe Gln Ser Phe Ser 545
550 555 560 Tyr Ala Thr Ile Asn Thr Ala Phe Thr Phe Pro Met Ser Gln
Ser Ser 565 570 575 Phe Thr Val Gly Ala Asp Thr Phe Ser Ser Gly Asn
Glu Val Tyr Ile 580 585 590 Asp Arg Phe Glu Leu Ile Pro Val Thr Ala
Thr Phe Glu 595 600 605 51818DNAArtificial SequenceSynthetic DNA
sequence in accordance with the invention encoding Bacillus
thuringiensis Cry1Fa core toxin using codons optimized for maize
and with sequences identified in Table 2 removed and Table 1
sequences are maintained 5atg gag aat aat atc cag aat caa tgc gtg
cct tac aat tgt ctc aat 48Met Glu Asn Asn Ile Gln Asn Gln Cys Val
Pro Tyr Asn Cys Leu Asn 1 5 10 15 aat ccc gag gtg gag ata tta aac
gag gag aga tcc act ggc aga ctg 96Asn Pro Glu Val Glu Ile Leu Asn
Glu Glu Arg Ser Thr Gly Arg Leu 20 25 30 cca ctc gac ata tcc ttg
tcc ctt acc cgt ttc ctt ttg agc gaa ttt 144Pro Leu Asp Ile Ser Leu
Ser Leu Thr Arg Phe Leu Leu Ser Glu Phe 35 40 45 gtt cct ggt gtg
gga gtg gct ttc gga ctg ttc gat ctg ata tgg ggc 192Val Pro Gly Val
Gly Val Ala Phe Gly Leu Phe Asp Leu Ile Trp Gly 50 55 60 ttt atc
act cct tct gat tgg agc ctc ttc ctt ctc cag att gag caa 240Phe Ile
Thr Pro Ser Asp Trp Ser Leu Phe Leu Leu Gln Ile Glu Gln 65 70 75 80
ttg att gag cag aga ata gaa acc ttg gaa agg aac cgt gca atc acg
288Leu Ile Glu Gln Arg Ile Glu Thr Leu Glu Arg Asn Arg Ala Ile Thr
85 90 95 acc ttg cgc ggt ctc gcc gat agc tat gaa atc tac att gaa
gca ctg 336Thr Leu Arg Gly Leu Ala Asp Ser Tyr Glu Ile Tyr Ile Glu
Ala Leu 100 105 110 agg gag tgg gag gcc aac ccc aat aat gct caa tta
agg gaa gat gtg 384Arg Glu Trp Glu Ala Asn Pro Asn Asn Ala Gln Leu
Arg Glu Asp Val 115 120 125 cgt att cgt ttt gct aat aca gac gac gct
ctc atc aca gca atc aat 432Arg Ile Arg Phe Ala Asn Thr Asp Asp Ala
Leu Ile Thr Ala Ile Asn 130 135 140 aat ttc aca ctt aca tcc ttt gaa
atc ccg ctt ttg agc gtg tac gtt 480Asn Phe Thr Leu Thr Ser Phe Glu
Ile Pro Leu Leu Ser Val Tyr Val 145 150 155 160 caa gcc gcc aat ctc
cac ctc tca ctt ctg agg gac gct gtc tcc ttt 528Gln Ala Ala Asn Leu
His Leu Ser Leu Leu Arg Asp Ala Val Ser Phe 165 170 175 ggg caa ggt
tgg gga ctg gat atc gct act gtg aat aat cac tac aat 576Gly Gln Gly
Trp Gly Leu Asp Ile Ala Thr Val Asn Asn His Tyr Asn 180 185 190 aga
tta atc aac ctg att cat aga tat acg aag cac tgc ttg gac aca 624Arg
Leu Ile Asn Leu Ile His Arg Tyr Thr Lys His Cys Leu Asp Thr 195 200
205 tac aat caa gga ctg gag aac ctt agg gga act aac act agg cag tgg
672Tyr Asn Gln Gly Leu Glu Asn Leu Arg Gly Thr Asn Thr Arg Gln Trp
210 215 220 gca agg ttc aac cag ttc aga cgt gat ctc aca ctt act gtg
ctg gat 720Ala Arg Phe Asn Gln Phe Arg Arg Asp Leu Thr Leu Thr Val
Leu Asp 225 230 235 240 atc gtt gct ctc ttt ccg aac tac gat gtt cgc
acc tac cca atc cag 768Ile Val Ala Leu Phe Pro Asn Tyr Asp Val Arg
Thr Tyr Pro Ile Gln 245 250 255 acg tca tcc caa tta aca agg gaa atc
tac acc tcc tca gtg att gag 816Thr Ser Ser Gln Leu Thr Arg Glu Ile
Tyr Thr Ser Ser Val Ile Glu 260 265 270 gac tct ccc gtt tct gct aac
ata cct aac ggc ttc aac cgc gcc gag 864Asp Ser Pro Val Ser Ala Asn
Ile Pro Asn Gly Phe Asn Arg Ala Glu 275 280 285 ttc gga gtt aga ccg
ccc cac ctt atg gac ttt atg aat agc ttg ttc 912Phe Gly Val Arg Pro
Pro His Leu Met Asp Phe Met Asn Ser Leu Phe 290 295 300 gtg act gct
gag act gtt aga agc caa act gtg tgg ggc ggc cac ttg 960Val Thr Ala
Glu Thr Val Arg Ser Gln Thr Val Trp Gly Gly His Leu 305 310 315 320
gtc agc tca cgc aac acg gct ggc aac cgt atc aac ttc ccg tct tac
1008Val Ser Ser Arg Asn Thr Ala Gly Asn Arg Ile Asn Phe Pro Ser Tyr
325 330 335 ggg gtc ttt aac cct ggt ggc gcc att tgg att gca gac gag
gac cca 1056Gly Val Phe Asn Pro Gly Gly Ala Ile Trp Ile Ala Asp Glu
Asp Pro 340 345 350 cgt cct ttt tac cgc acc ctg tca gat ccg gtt ttc
gtc aga ggc gga 1104Arg Pro Phe Tyr Arg Thr Leu Ser Asp Pro Val Phe
Val Arg Gly Gly 355 360 365 ttt ggg aat cct cat tat gtc ctg ggc ctt
agg gga gtg gct ttc caa 1152Phe Gly Asn Pro His Tyr Val Leu Gly Leu
Arg Gly Val Ala Phe Gln 370 375 380 cag act ggc acc aac cac acc cgt
acg ttt cgc aat agc ggg acc ata 1200Gln Thr Gly Thr Asn His Thr Arg
Thr Phe Arg Asn Ser Gly Thr Ile 385 390 395 400 gat tct ctt gat gaa
atc cca cct caa gat aac agc ggc gca cct tgg 1248Asp Ser Leu Asp Glu
Ile Pro Pro Gln Asp Asn Ser Gly Ala Pro Trp 405 410 415 aac gat tat
tcc cac gta tta aat cac gtt acg ttc gtc cgc tgg ccg 1296Asn Asp Tyr
Ser His Val Leu Asn His Val Thr Phe Val Arg Trp Pro 420 425 430 ggt
gag atc agc ggc agc gat tca tgg aga gca cca atg ttc tct tgg 1344Gly
Glu Ile Ser Gly Ser Asp Ser Trp Arg Ala Pro Met Phe Ser Trp 435 440
445 acg cac cgt tca gcc acc cct aca aat aca att gac ccg gag agg att
1392Thr His Arg Ser Ala Thr Pro Thr Asn Thr Ile Asp Pro Glu Arg Ile
450 455 460 act caa atc cca ttg gtc aaa gca cat aca ctt cag tct ggg
acc acc 1440Thr Gln Ile Pro Leu Val Lys Ala His Thr Leu Gln Ser Gly
Thr Thr 465 470 475 480 gtg gtc aga ggg cct ggg ttc acg gga gga gac
att ctt agg cgc aca 1488Val Val Arg Gly Pro Gly Phe Thr Gly Gly Asp
Ile Leu Arg Arg Thr 485 490 495 tcc gga gga ccc ttc gct tat act atc
gtt aat ata aat ggg cag ctc 1536Ser Gly Gly Pro Phe Ala Tyr Thr Ile
Val Asn Ile Asn Gly Gln Leu 500 505 510 ccc cag cgc tat cgt gcc aga
atc cgt tac gcc tct act aca aat ctc 1584Pro Gln Arg Tyr Arg Ala Arg
Ile Arg Tyr Ala Ser Thr Thr Asn Leu 515 520 525 aga atc tac gtg acg
gtt gcc ggt gag cgc atc ttt gct ggt cag ttt 1632Arg Ile Tyr Val Thr
Val Ala Gly Glu Arg Ile Phe Ala Gly Gln Phe 530 535 540 aac aag acg
atg gat act ggc gac cca ctg aca ttc caa tct ttc tca 1680Asn Lys Thr
Met Asp Thr Gly Asp Pro Leu Thr Phe Gln Ser Phe Ser 545 550 555 560
tac gca act att aat aca gct ttc aca ttc cca atg agc cag tca tct
1728Tyr Ala Thr Ile Asn Thr Ala Phe Thr Phe Pro Met Ser Gln Ser Ser
565 570 575 ttc acc gtc ggt gct gat acc ttc agc tct ggc aac gaa gtc
tat atc 1776Phe Thr Val Gly Ala Asp Thr Phe Ser Ser Gly Asn Glu Val
Tyr Ile 580 585 590 gac aga ttt gag ttg att cca gtt act gca acg ttt
gag tga 1818Asp Arg Phe Glu Leu Ile Pro Val Thr Ala Thr Phe Glu 595
600 605 6605PRTArtificial SequenceSynthetic Construct 6Met Glu Asn
Asn Ile Gln Asn Gln Cys Val Pro Tyr Asn Cys Leu Asn 1 5 10 15 Asn
Pro Glu Val Glu Ile Leu Asn Glu Glu Arg Ser Thr Gly Arg Leu 20 25
30 Pro Leu Asp Ile Ser Leu Ser Leu Thr Arg Phe Leu Leu Ser Glu Phe
35 40 45 Val Pro Gly Val Gly Val Ala Phe Gly Leu Phe Asp Leu Ile
Trp Gly 50 55 60 Phe Ile Thr Pro Ser Asp Trp Ser Leu Phe Leu Leu
Gln Ile Glu Gln 65 70 75 80 Leu Ile Glu Gln Arg Ile Glu Thr Leu Glu
Arg Asn Arg Ala Ile Thr 85 90 95 Thr Leu Arg Gly Leu Ala Asp Ser
Tyr Glu Ile Tyr Ile Glu Ala Leu 100 105 110 Arg Glu Trp Glu Ala Asn
Pro Asn Asn Ala Gln Leu Arg Glu Asp Val 115 120 125 Arg Ile Arg Phe
Ala Asn Thr Asp Asp Ala Leu Ile Thr Ala Ile Asn 130 135 140 Asn Phe
Thr Leu Thr Ser Phe Glu Ile Pro Leu Leu Ser Val Tyr Val 145 150 155
160 Gln Ala Ala Asn Leu His Leu Ser Leu Leu Arg Asp Ala Val Ser Phe
165 170 175 Gly Gln Gly Trp Gly Leu Asp Ile Ala Thr Val Asn Asn His
Tyr Asn 180
185 190 Arg Leu Ile Asn Leu Ile His Arg Tyr Thr Lys His Cys Leu Asp
Thr 195 200 205 Tyr Asn Gln Gly Leu Glu Asn Leu Arg Gly Thr Asn Thr
Arg Gln Trp 210 215 220 Ala Arg Phe Asn Gln Phe Arg Arg Asp Leu Thr
Leu Thr Val Leu Asp 225 230 235 240 Ile Val Ala Leu Phe Pro Asn Tyr
Asp Val Arg Thr Tyr Pro Ile Gln 245 250 255 Thr Ser Ser Gln Leu Thr
Arg Glu Ile Tyr Thr Ser Ser Val Ile Glu 260 265 270 Asp Ser Pro Val
Ser Ala Asn Ile Pro Asn Gly Phe Asn Arg Ala Glu 275 280 285 Phe Gly
Val Arg Pro Pro His Leu Met Asp Phe Met Asn Ser Leu Phe 290 295 300
Val Thr Ala Glu Thr Val Arg Ser Gln Thr Val Trp Gly Gly His Leu 305
310 315 320 Val Ser Ser Arg Asn Thr Ala Gly Asn Arg Ile Asn Phe Pro
Ser Tyr 325 330 335 Gly Val Phe Asn Pro Gly Gly Ala Ile Trp Ile Ala
Asp Glu Asp Pro 340 345 350 Arg Pro Phe Tyr Arg Thr Leu Ser Asp Pro
Val Phe Val Arg Gly Gly 355 360 365 Phe Gly Asn Pro His Tyr Val Leu
Gly Leu Arg Gly Val Ala Phe Gln 370 375 380 Gln Thr Gly Thr Asn His
Thr Arg Thr Phe Arg Asn Ser Gly Thr Ile 385 390 395 400 Asp Ser Leu
Asp Glu Ile Pro Pro Gln Asp Asn Ser Gly Ala Pro Trp 405 410 415 Asn
Asp Tyr Ser His Val Leu Asn His Val Thr Phe Val Arg Trp Pro 420 425
430 Gly Glu Ile Ser Gly Ser Asp Ser Trp Arg Ala Pro Met Phe Ser Trp
435 440 445 Thr His Arg Ser Ala Thr Pro Thr Asn Thr Ile Asp Pro Glu
Arg Ile 450 455 460 Thr Gln Ile Pro Leu Val Lys Ala His Thr Leu Gln
Ser Gly Thr Thr 465 470 475 480 Val Val Arg Gly Pro Gly Phe Thr Gly
Gly Asp Ile Leu Arg Arg Thr 485 490 495 Ser Gly Gly Pro Phe Ala Tyr
Thr Ile Val Asn Ile Asn Gly Gln Leu 500 505 510 Pro Gln Arg Tyr Arg
Ala Arg Ile Arg Tyr Ala Ser Thr Thr Asn Leu 515 520 525 Arg Ile Tyr
Val Thr Val Ala Gly Glu Arg Ile Phe Ala Gly Gln Phe 530 535 540 Asn
Lys Thr Met Asp Thr Gly Asp Pro Leu Thr Phe Gln Ser Phe Ser 545 550
555 560 Tyr Ala Thr Ile Asn Thr Ala Phe Thr Phe Pro Met Ser Gln Ser
Ser 565 570 575 Phe Thr Val Gly Ala Asp Thr Phe Ser Ser Gly Asn Glu
Val Tyr Ile 580 585 590 Asp Arg Phe Glu Leu Ile Pro Val Thr Ala Thr
Phe Glu 595 600 605 7372DNABacillus
thuringiensisCDS(1)..(372)Native DNA sequence encoding Bacillus
thuringiensis Cry34Ab1 toxin 7atg tca gca cgt gaa gta cac att gat
gta aat aat aag aca ggt cat 48Met Ser Ala Arg Glu Val His Ile Asp
Val Asn Asn Lys Thr Gly His 1 5 10 15 aca tta caa tta gaa gat aaa
aca aaa ctt gat ggt ggt aga tgg cga 96Thr Leu Gln Leu Glu Asp Lys
Thr Lys Leu Asp Gly Gly Arg Trp Arg 20 25 30 aca tca cct aca aat
gtt gct aat gat caa att aaa aca ttt gta gca 144Thr Ser Pro Thr Asn
Val Ala Asn Asp Gln Ile Lys Thr Phe Val Ala 35 40 45 gaa tca aat
ggt ttt atg aca ggt aca gaa ggt act ata tat tat agt 192Glu Ser Asn
Gly Phe Met Thr Gly Thr Glu Gly Thr Ile Tyr Tyr Ser 50 55 60 ata
aat gga gaa gca gaa att agt tta tat ttt gac aat cct ttt gca 240Ile
Asn Gly Glu Ala Glu Ile Ser Leu Tyr Phe Asp Asn Pro Phe Ala 65 70
75 80 ggt tct aat aaa tat gat gga cat tcc aat aaa tct caa tat gaa
att 288Gly Ser Asn Lys Tyr Asp Gly His Ser Asn Lys Ser Gln Tyr Glu
Ile 85 90 95 att acc caa gga gga tca gga aat caa tct cat gtt acg
tat act att 336Ile Thr Gln Gly Gly Ser Gly Asn Gln Ser His Val Thr
Tyr Thr Ile 100 105 110 caa acc aca tcc tca cga tat ggg cat aaa tca
taa 372Gln Thr Thr Ser Ser Arg Tyr Gly His Lys Ser 115 120
8123PRTBacillus thuringiensis 8Met Ser Ala Arg Glu Val His Ile Asp
Val Asn Asn Lys Thr Gly His 1 5 10 15 Thr Leu Gln Leu Glu Asp Lys
Thr Lys Leu Asp Gly Gly Arg Trp Arg 20 25 30 Thr Ser Pro Thr Asn
Val Ala Asn Asp Gln Ile Lys Thr Phe Val Ala 35 40 45 Glu Ser Asn
Gly Phe Met Thr Gly Thr Glu Gly Thr Ile Tyr Tyr Ser 50 55 60 Ile
Asn Gly Glu Ala Glu Ile Ser Leu Tyr Phe Asp Asn Pro Phe Ala 65 70
75 80 Gly Ser Asn Lys Tyr Asp Gly His Ser Asn Lys Ser Gln Tyr Glu
Ile 85 90 95 Ile Thr Gln Gly Gly Ser Gly Asn Gln Ser His Val Thr
Tyr Thr Ile 100 105 110 Gln Thr Thr Ser Ser Arg Tyr Gly His Lys Ser
115 120 9372DNAArtificial SequenceSynthetic DNA sequence encoding
Bacillus thuringiensis Cry34Ab1 toxin using codons optimized for
maize and Table 1 sequences are maintained 9atg tca gca cgg gag gtc
cac atc gat gta aat aat aag acg ggt cat 48Met Ser Ala Arg Glu Val
His Ile Asp Val Asn Asn Lys Thr Gly His 1 5 10 15 aca tta cag ttg
gag gat aaa aca aag cta gac ggt ggc aga tgg aga 96Thr Leu Gln Leu
Glu Asp Lys Thr Lys Leu Asp Gly Gly Arg Trp Arg 20 25 30 acc agt
ccg acc aac gtt gct aac gat caa att aaa aca ttt gta gcc 144Thr Ser
Pro Thr Asn Val Ala Asn Asp Gln Ile Lys Thr Phe Val Ala 35 40 45
gaa tca aac ggt ttt atg act ggc acg gag ggg act ata tat tat tcc
192Glu Ser Asn Gly Phe Met Thr Gly Thr Glu Gly Thr Ile Tyr Tyr Ser
50 55 60 atc aac gga gaa gcc gag att tcg tta tat ttt gac aat cca
ttc gcg 240Ile Asn Gly Glu Ala Glu Ile Ser Leu Tyr Phe Asp Asn Pro
Phe Ala 65 70 75 80 ggg tct aat aaa tac gac gga cac tcc aat aaa tct
caa tat gaa atc 288Gly Ser Asn Lys Tyr Asp Gly His Ser Asn Lys Ser
Gln Tyr Glu Ile 85 90 95 att aca caa ggc ggc agc gga aat caa agc
cac gtc acg tat act atc 336Ile Thr Gln Gly Gly Ser Gly Asn Gln Ser
His Val Thr Tyr Thr Ile 100 105 110 cag acc act tca tcg cgc tac ggg
cat aaa tca tag 372Gln Thr Thr Ser Ser Arg Tyr Gly His Lys Ser 115
120 10123PRTArtificial SequenceSynthetic Construct 10Met Ser Ala
Arg Glu Val His Ile Asp Val Asn Asn Lys Thr Gly His 1 5 10 15 Thr
Leu Gln Leu Glu Asp Lys Thr Lys Leu Asp Gly Gly Arg Trp Arg 20 25
30 Thr Ser Pro Thr Asn Val Ala Asn Asp Gln Ile Lys Thr Phe Val Ala
35 40 45 Glu Ser Asn Gly Phe Met Thr Gly Thr Glu Gly Thr Ile Tyr
Tyr Ser 50 55 60 Ile Asn Gly Glu Ala Glu Ile Ser Leu Tyr Phe Asp
Asn Pro Phe Ala 65 70 75 80 Gly Ser Asn Lys Tyr Asp Gly His Ser Asn
Lys Ser Gln Tyr Glu Ile 85 90 95 Ile Thr Gln Gly Gly Ser Gly Asn
Gln Ser His Val Thr Tyr Thr Ile 100 105 110 Gln Thr Thr Ser Ser Arg
Tyr Gly His Lys Ser 115 120 11372DNAArtificial SequenceSynthetic
DNA sequence in accordance with the invention encoding Bacillus
thuringiensis Cry34Ab1 toxin using codons optimized for maize and
with sequences identified in Table 2 removed and Table 1 sequences
are maintained 11atg tca gca cgg gag gtc cac atc gat gta aat aat
aag acg ggt cat 48Met Ser Ala Arg Glu Val His Ile Asp Val Asn Asn
Lys Thr Gly His 1 5 10 15 aca tta cag ttg gag gat aaa aca aag cta
gac ggt ggc aga tgg aga 96Thr Leu Gln Leu Glu Asp Lys Thr Lys Leu
Asp Gly Gly Arg Trp Arg 20 25 30 acc agt ccg acc aac gtt gct aac
gat caa att aaa aca ttt gta gcc 144Thr Ser Pro Thr Asn Val Ala Asn
Asp Gln Ile Lys Thr Phe Val Ala 35 40 45 gaa tca aac ggt ttc atg
act ggc acg gag ggg act atc tac tac tcc 192Glu Ser Asn Gly Phe Met
Thr Gly Thr Glu Gly Thr Ile Tyr Tyr Ser 50 55 60 atc aac gga gaa
gcc gag att tcg ctg tac ttc gac aat cca ttc gcg 240Ile Asn Gly Glu
Ala Glu Ile Ser Leu Tyr Phe Asp Asn Pro Phe Ala 65 70 75 80 ggg tct
aat aaa tac gac gga cac tcc aat aaa tct caa tat gaa atc 288Gly Ser
Asn Lys Tyr Asp Gly His Ser Asn Lys Ser Gln Tyr Glu Ile 85 90 95
att aca caa ggc ggc agc gga aat caa agc cac gtc acg tat act atc
336Ile Thr Gln Gly Gly Ser Gly Asn Gln Ser His Val Thr Tyr Thr Ile
100 105 110 cag acc act tca tcg cgc tac ggg cat aaa tca tag 372Gln
Thr Thr Ser Ser Arg Tyr Gly His Lys Ser 115 120 12123PRTArtificial
SequenceSynthetic Construct 12Met Ser Ala Arg Glu Val His Ile Asp
Val Asn Asn Lys Thr Gly His 1 5 10 15 Thr Leu Gln Leu Glu Asp Lys
Thr Lys Leu Asp Gly Gly Arg Trp Arg 20 25 30 Thr Ser Pro Thr Asn
Val Ala Asn Asp Gln Ile Lys Thr Phe Val Ala 35 40 45 Glu Ser Asn
Gly Phe Met Thr Gly Thr Glu Gly Thr Ile Tyr Tyr Ser 50 55 60 Ile
Asn Gly Glu Ala Glu Ile Ser Leu Tyr Phe Asp Asn Pro Phe Ala 65 70
75 80 Gly Ser Asn Lys Tyr Asp Gly His Ser Asn Lys Ser Gln Tyr Glu
Ile 85 90 95 Ile Thr Gln Gly Gly Ser Gly Asn Gln Ser His Val Thr
Tyr Thr Ile 100 105 110 Gln Thr Thr Ser Ser Arg Tyr Gly His Lys Ser
115 120 131152DNABacillus thuringiensisCDS(1)..(1152)Native DNA
sequence encoding Bacillus thuringiensis Cry35Ab1 toxin 13atg tta
gat act aat aaa gtt tat gaa ata agc aat cat gct aat gga 48Met Leu
Asp Thr Asn Lys Val Tyr Glu Ile Ser Asn His Ala Asn Gly 1 5 10 15
cta tat gca gca act tat tta agt tta gat gat tca ggt gtt agt tta
96Leu Tyr Ala Ala Thr Tyr Leu Ser Leu Asp Asp Ser Gly Val Ser Leu
20 25 30 atg aat aaa aat gat gat gat att gat gat tat aac tta aaa
tgg ttt 144Met Asn Lys Asn Asp Asp Asp Ile Asp Asp Tyr Asn Leu Lys
Trp Phe 35 40 45 tta ttt cct att gat gat gat caa tat att att aca
agc tat gca gca 192Leu Phe Pro Ile Asp Asp Asp Gln Tyr Ile Ile Thr
Ser Tyr Ala Ala 50 55 60 aat aat tgt aaa gtt tgg aat gtt aat aat
gat aaa ata aat gtt tcg 240Asn Asn Cys Lys Val Trp Asn Val Asn Asn
Asp Lys Ile Asn Val Ser 65 70 75 80 act tat tct tca aca aat tca ata
caa aaa tgg caa ata aaa gct aat 288Thr Tyr Ser Ser Thr Asn Ser Ile
Gln Lys Trp Gln Ile Lys Ala Asn 85 90 95 ggt tct tca tat gta ata
caa agt gat aat gga aaa gtc tta aca gca 336Gly Ser Ser Tyr Val Ile
Gln Ser Asp Asn Gly Lys Val Leu Thr Ala 100 105 110 gga acc ggt caa
gct ctt gga ttg ata cgt tta act gat gaa tcc tca 384Gly Thr Gly Gln
Ala Leu Gly Leu Ile Arg Leu Thr Asp Glu Ser Ser 115 120 125 aat aat
ccc aat caa caa tgg aat tta act tct gta caa aca att caa 432Asn Asn
Pro Asn Gln Gln Trp Asn Leu Thr Ser Val Gln Thr Ile Gln 130 135 140
ctt cca caa aaa cct ata ata gat aca aaa tta aaa gat tat ccc aaa
480Leu Pro Gln Lys Pro Ile Ile Asp Thr Lys Leu Lys Asp Tyr Pro Lys
145 150 155 160 tat tca cca act gga aat ata gat aat gga aca tct cct
caa tta atg 528Tyr Ser Pro Thr Gly Asn Ile Asp Asn Gly Thr Ser Pro
Gln Leu Met 165 170 175 gga tgg aca tta gta cct tgt att atg gta aat
gat cca aat ata gat 576Gly Trp Thr Leu Val Pro Cys Ile Met Val Asn
Asp Pro Asn Ile Asp 180 185 190 aaa aat act caa att aaa act act cca
tat tat att tta aaa aaa tat 624Lys Asn Thr Gln Ile Lys Thr Thr Pro
Tyr Tyr Ile Leu Lys Lys Tyr 195 200 205 caa tat tgg caa cga gca gta
gga agt aat gta gct tta cgt cca cat 672Gln Tyr Trp Gln Arg Ala Val
Gly Ser Asn Val Ala Leu Arg Pro His 210 215 220 gaa aaa aaa tca tat
act tat gaa tgg ggc aca gaa ata gat caa aaa 720Glu Lys Lys Ser Tyr
Thr Tyr Glu Trp Gly Thr Glu Ile Asp Gln Lys 225 230 235 240 aca aca
att ata aat aca tta gga ttt caa atc aat ata gat tca gga 768Thr Thr
Ile Ile Asn Thr Leu Gly Phe Gln Ile Asn Ile Asp Ser Gly 245 250 255
atg aaa ttt gat ata cca gaa gta ggt gga ggt aca gat gaa ata aaa
816Met Lys Phe Asp Ile Pro Glu Val Gly Gly Gly Thr Asp Glu Ile Lys
260 265 270 aca caa cta aat gaa gaa tta aaa ata gaa tat agt cat gaa
act aaa 864Thr Gln Leu Asn Glu Glu Leu Lys Ile Glu Tyr Ser His Glu
Thr Lys 275 280 285 ata atg gaa aaa tat caa gaa caa tct gaa ata gat
aat cca act gat 912Ile Met Glu Lys Tyr Gln Glu Gln Ser Glu Ile Asp
Asn Pro Thr Asp 290 295 300 caa tca atg aat tct ata gga ttt ctt act
att act tcc tta gaa tta 960Gln Ser Met Asn Ser Ile Gly Phe Leu Thr
Ile Thr Ser Leu Glu Leu 305 310 315 320 tat aga tat aat ggc tca gaa
att cgt ata atg caa att caa acc tca 1008Tyr Arg Tyr Asn Gly Ser Glu
Ile Arg Ile Met Gln Ile Gln Thr Ser 325 330 335 gat aat gat act tat
aat gtt act tct tat cca aat cat caa caa gct 1056Asp Asn Asp Thr Tyr
Asn Val Thr Ser Tyr Pro Asn His Gln Gln Ala 340 345 350 tta tta ctt
ctt aca aat cat tca tat gaa gaa gta gaa gaa ata aca 1104Leu Leu Leu
Leu Thr Asn His Ser Tyr Glu Glu Val Glu Glu Ile Thr 355 360 365 aat
att cct aaa agt aca cta aaa aaa tta aaa aaa tat tat ttt taa 1152Asn
Ile Pro Lys Ser Thr Leu Lys Lys Leu Lys Lys Tyr Tyr Phe 370 375 380
14383PRTBacillus thuringiensis 14Met Leu Asp Thr Asn Lys Val Tyr
Glu Ile Ser Asn His Ala Asn Gly 1 5 10 15 Leu Tyr Ala Ala Thr Tyr
Leu Ser Leu Asp Asp Ser Gly Val Ser Leu 20 25 30 Met Asn Lys Asn
Asp Asp Asp Ile Asp Asp Tyr Asn Leu Lys Trp Phe 35 40 45 Leu Phe
Pro Ile Asp Asp Asp Gln Tyr Ile Ile Thr Ser Tyr Ala Ala 50 55 60
Asn Asn Cys Lys Val Trp Asn Val Asn Asn Asp Lys Ile Asn Val Ser 65
70 75 80 Thr Tyr Ser Ser Thr Asn Ser Ile Gln Lys Trp Gln Ile Lys
Ala Asn 85 90 95 Gly Ser
Ser Tyr Val Ile Gln Ser Asp Asn Gly Lys Val Leu Thr Ala 100 105 110
Gly Thr Gly Gln Ala Leu Gly Leu Ile Arg Leu Thr Asp Glu Ser Ser 115
120 125 Asn Asn Pro Asn Gln Gln Trp Asn Leu Thr Ser Val Gln Thr Ile
Gln 130 135 140 Leu Pro Gln Lys Pro Ile Ile Asp Thr Lys Leu Lys Asp
Tyr Pro Lys 145 150 155 160 Tyr Ser Pro Thr Gly Asn Ile Asp Asn Gly
Thr Ser Pro Gln Leu Met 165 170 175 Gly Trp Thr Leu Val Pro Cys Ile
Met Val Asn Asp Pro Asn Ile Asp 180 185 190 Lys Asn Thr Gln Ile Lys
Thr Thr Pro Tyr Tyr Ile Leu Lys Lys Tyr 195 200 205 Gln Tyr Trp Gln
Arg Ala Val Gly Ser Asn Val Ala Leu Arg Pro His 210 215 220 Glu Lys
Lys Ser Tyr Thr Tyr Glu Trp Gly Thr Glu Ile Asp Gln Lys 225 230 235
240 Thr Thr Ile Ile Asn Thr Leu Gly Phe Gln Ile Asn Ile Asp Ser Gly
245 250 255 Met Lys Phe Asp Ile Pro Glu Val Gly Gly Gly Thr Asp Glu
Ile Lys 260 265 270 Thr Gln Leu Asn Glu Glu Leu Lys Ile Glu Tyr Ser
His Glu Thr Lys 275 280 285 Ile Met Glu Lys Tyr Gln Glu Gln Ser Glu
Ile Asp Asn Pro Thr Asp 290 295 300 Gln Ser Met Asn Ser Ile Gly Phe
Leu Thr Ile Thr Ser Leu Glu Leu 305 310 315 320 Tyr Arg Tyr Asn Gly
Ser Glu Ile Arg Ile Met Gln Ile Gln Thr Ser 325 330 335 Asp Asn Asp
Thr Tyr Asn Val Thr Ser Tyr Pro Asn His Gln Gln Ala 340 345 350 Leu
Leu Leu Leu Thr Asn His Ser Tyr Glu Glu Val Glu Glu Ile Thr 355 360
365 Asn Ile Pro Lys Ser Thr Leu Lys Lys Leu Lys Lys Tyr Tyr Phe 370
375 380 151152DNAArtificial SequenceSynthetic DNA sequence encoding
Bacillus thuringiensis Cry35Ab1 toxin using codons optimized for
maize and Table 1 sequences are maintained 15atg ctc gat act aat
aaa gtg tat gaa ata agc aac cat gcc aac ggg 48Met Leu Asp Thr Asn
Lys Val Tyr Glu Ile Ser Asn His Ala Asn Gly 1 5 10 15 cta tat gcc
gca act tat ttg agt ctg gac gac agc ggt gtg agc tta 96Leu Tyr Ala
Ala Thr Tyr Leu Ser Leu Asp Asp Ser Gly Val Ser Leu 20 25 30 atg
aat aaa aac gac gac gac att gac gac tac aac ctc aag tgg ttt 144Met
Asn Lys Asn Asp Asp Asp Ile Asp Asp Tyr Asn Leu Lys Trp Phe 35 40
45 tta ttt cct att gac gac gat caa tat att att aca agc tac gca gca
192Leu Phe Pro Ile Asp Asp Asp Gln Tyr Ile Ile Thr Ser Tyr Ala Ala
50 55 60 aat aat tgc aaa gtc tgg aac gtt aat aat gat aaa ata aat
gtt tcg 240Asn Asn Cys Lys Val Trp Asn Val Asn Asn Asp Lys Ile Asn
Val Ser 65 70 75 80 acc tac agc tcc acc aac tca ata caa aag tgg caa
ata aaa gct aat 288Thr Tyr Ser Ser Thr Asn Ser Ile Gln Lys Trp Gln
Ile Lys Ala Asn 85 90 95 gga tcg tcg tat gta ata cag agt gac aat
ggg aag gtc ttg aca gcg 336Gly Ser Ser Tyr Val Ile Gln Ser Asp Asn
Gly Lys Val Leu Thr Ala 100 105 110 ggc act ggt caa gct ctt gga ctc
ata agg ctc act gac gag tcc tca 384Gly Thr Gly Gln Ala Leu Gly Leu
Ile Arg Leu Thr Asp Glu Ser Ser 115 120 125 aat aat ccc aat caa cag
tgg aac ttg act tcc gtg cag acg atc caa 432Asn Asn Pro Asn Gln Gln
Trp Asn Leu Thr Ser Val Gln Thr Ile Gln 130 135 140 ctt cca cag aaa
cct atc atc gat aca aaa tta aaa gat tac ccc aag 480Leu Pro Gln Lys
Pro Ile Ile Asp Thr Lys Leu Lys Asp Tyr Pro Lys 145 150 155 160 tac
tcg cca acc ggc aac atc gat aat gga acg tct cct caa tta atg 528Tyr
Ser Pro Thr Gly Asn Ile Asp Asn Gly Thr Ser Pro Gln Leu Met 165 170
175 ggc tgg acc ctc gta ccc tgt att atg gtg aac gac ccg aat atc gat
576Gly Trp Thr Leu Val Pro Cys Ile Met Val Asn Asp Pro Asn Ile Asp
180 185 190 aaa aat act caa att aaa acc acg ccg tat tat ata ttg aaa
aaa tac 624Lys Asn Thr Gln Ile Lys Thr Thr Pro Tyr Tyr Ile Leu Lys
Lys Tyr 195 200 205 caa tac tgg cag cgc gcg gtt ggc tca aac gtc gct
ctg cgg cca cat 672Gln Tyr Trp Gln Arg Ala Val Gly Ser Asn Val Ala
Leu Arg Pro His 210 215 220 gaa aag aag tcc tac act tac gaa tgg ggc
aca gag atc gat cag aaa 720Glu Lys Lys Ser Tyr Thr Tyr Glu Trp Gly
Thr Glu Ile Asp Gln Lys 225 230 235 240 acg acc att ata aat aca tta
gga ttc caa atc aat atc gac agc gga 768Thr Thr Ile Ile Asn Thr Leu
Gly Phe Gln Ile Asn Ile Asp Ser Gly 245 250 255 atg aaa ttt gac atc
ccg gaa gtg ggg ggc ggg acc gat gaa ata aaa 816Met Lys Phe Asp Ile
Pro Glu Val Gly Gly Gly Thr Asp Glu Ile Lys 260 265 270 acg cag ctc
aac gaa gaa tta aaa ata gag tac agt cat gaa act aaa 864Thr Gln Leu
Asn Glu Glu Leu Lys Ile Glu Tyr Ser His Glu Thr Lys 275 280 285 ata
atg gaa aaa tat caa gag caa tct gaa atc gat aac ccg acc gac 912Ile
Met Glu Lys Tyr Gln Glu Gln Ser Glu Ile Asp Asn Pro Thr Asp 290 295
300 caa tca atg aac tct atc ggt ttc ctt act att acc tcc ctg gag tta
960Gln Ser Met Asn Ser Ile Gly Phe Leu Thr Ile Thr Ser Leu Glu Leu
305 310 315 320 tat aga tat aac ggc tct gag atc cgt ata atg cag att
caa acc tca 1008Tyr Arg Tyr Asn Gly Ser Glu Ile Arg Ile Met Gln Ile
Gln Thr Ser 325 330 335 gac aat gac act tat aac gtc acc tct tac ccg
aat cat caa caa gct 1056Asp Asn Asp Thr Tyr Asn Val Thr Ser Tyr Pro
Asn His Gln Gln Ala 340 345 350 tta ttg ctt ctt aca aac cac agt tat
gaa gag gtg gaa gaa ata acg 1104Leu Leu Leu Leu Thr Asn His Ser Tyr
Glu Glu Val Glu Glu Ile Thr 355 360 365 aac att cct aaa tcc aca cta
aag aaa tta aaa aaa tat tat ttt tga 1152Asn Ile Pro Lys Ser Thr Leu
Lys Lys Leu Lys Lys Tyr Tyr Phe 370 375 380 16383PRTArtificial
SequenceSynthetic Construct 16Met Leu Asp Thr Asn Lys Val Tyr Glu
Ile Ser Asn His Ala Asn Gly 1 5 10 15 Leu Tyr Ala Ala Thr Tyr Leu
Ser Leu Asp Asp Ser Gly Val Ser Leu 20 25 30 Met Asn Lys Asn Asp
Asp Asp Ile Asp Asp Tyr Asn Leu Lys Trp Phe 35 40 45 Leu Phe Pro
Ile Asp Asp Asp Gln Tyr Ile Ile Thr Ser Tyr Ala Ala 50 55 60 Asn
Asn Cys Lys Val Trp Asn Val Asn Asn Asp Lys Ile Asn Val Ser 65 70
75 80 Thr Tyr Ser Ser Thr Asn Ser Ile Gln Lys Trp Gln Ile Lys Ala
Asn 85 90 95 Gly Ser Ser Tyr Val Ile Gln Ser Asp Asn Gly Lys Val
Leu Thr Ala 100 105 110 Gly Thr Gly Gln Ala Leu Gly Leu Ile Arg Leu
Thr Asp Glu Ser Ser 115 120 125 Asn Asn Pro Asn Gln Gln Trp Asn Leu
Thr Ser Val Gln Thr Ile Gln 130 135 140 Leu Pro Gln Lys Pro Ile Ile
Asp Thr Lys Leu Lys Asp Tyr Pro Lys 145 150 155 160 Tyr Ser Pro Thr
Gly Asn Ile Asp Asn Gly Thr Ser Pro Gln Leu Met 165 170 175 Gly Trp
Thr Leu Val Pro Cys Ile Met Val Asn Asp Pro Asn Ile Asp 180 185 190
Lys Asn Thr Gln Ile Lys Thr Thr Pro Tyr Tyr Ile Leu Lys Lys Tyr 195
200 205 Gln Tyr Trp Gln Arg Ala Val Gly Ser Asn Val Ala Leu Arg Pro
His 210 215 220 Glu Lys Lys Ser Tyr Thr Tyr Glu Trp Gly Thr Glu Ile
Asp Gln Lys 225 230 235 240 Thr Thr Ile Ile Asn Thr Leu Gly Phe Gln
Ile Asn Ile Asp Ser Gly 245 250 255 Met Lys Phe Asp Ile Pro Glu Val
Gly Gly Gly Thr Asp Glu Ile Lys 260 265 270 Thr Gln Leu Asn Glu Glu
Leu Lys Ile Glu Tyr Ser His Glu Thr Lys 275 280 285 Ile Met Glu Lys
Tyr Gln Glu Gln Ser Glu Ile Asp Asn Pro Thr Asp 290 295 300 Gln Ser
Met Asn Ser Ile Gly Phe Leu Thr Ile Thr Ser Leu Glu Leu 305 310 315
320 Tyr Arg Tyr Asn Gly Ser Glu Ile Arg Ile Met Gln Ile Gln Thr Ser
325 330 335 Asp Asn Asp Thr Tyr Asn Val Thr Ser Tyr Pro Asn His Gln
Gln Ala 340 345 350 Leu Leu Leu Leu Thr Asn His Ser Tyr Glu Glu Val
Glu Glu Ile Thr 355 360 365 Asn Ile Pro Lys Ser Thr Leu Lys Lys Leu
Lys Lys Tyr Tyr Phe 370 375 380 171152DNAArtificial
SequenceSynthetic DNA sequence in accordance with the invention
encoding Bacillus thuringiensis Cry35Ab1 toxin using codons
optimized for maize and with sequences identified in Table 2
removed and Table 1 sequences are maintained 17atg ctc gat act aat
aaa gtg tat gaa ata tcg aac cat gcc aac ggg 48Met Leu Asp Thr Asn
Lys Val Tyr Glu Ile Ser Asn His Ala Asn Gly 1 5 10 15 cta tat gcc
gca act tac ctg agt ctg gac gat agt ggt gtg agc tta 96Leu Tyr Ala
Ala Thr Tyr Leu Ser Leu Asp Asp Ser Gly Val Ser Leu 20 25 30 atg
aat aaa aac gac gac gac att gac gac tac aac ctc aag tgg ttc 144Met
Asn Lys Asn Asp Asp Asp Ile Asp Asp Tyr Asn Leu Lys Trp Phe 35 40
45 ctg ttt cct att gac gac gat cag tat atc att aca agc tac gca gcg
192Leu Phe Pro Ile Asp Asp Asp Gln Tyr Ile Ile Thr Ser Tyr Ala Ala
50 55 60 aat aat tgc aaa gtc tgg aac gtc aat aat gat aaa atc aat
gtt tcg 240Asn Asn Cys Lys Val Trp Asn Val Asn Asn Asp Lys Ile Asn
Val Ser 65 70 75 80 acc tac agc tcc acc aac tca ata caa aag tgg caa
atc aaa gct aat 288Thr Tyr Ser Ser Thr Asn Ser Ile Gln Lys Trp Gln
Ile Lys Ala Asn 85 90 95 ggc agc tcg tac gta ata cag agt gac aat
ggg aag gtc ttg aca gcg 336Gly Ser Ser Tyr Val Ile Gln Ser Asp Asn
Gly Lys Val Leu Thr Ala 100 105 110 ggc act ggt caa gct ctt gga ctc
ata agg ctc act gac gag tcc tcg 384Gly Thr Gly Gln Ala Leu Gly Leu
Ile Arg Leu Thr Asp Glu Ser Ser 115 120 125 aat aat ccc aat caa cag
tgg aac ttg act tcc gtg cag acg atc caa 432Asn Asn Pro Asn Gln Gln
Trp Asn Leu Thr Ser Val Gln Thr Ile Gln 130 135 140 ctt cca cag aaa
cct atc atc gat aca aaa tta aaa gat tac ccc aag 480Leu Pro Gln Lys
Pro Ile Ile Asp Thr Lys Leu Lys Asp Tyr Pro Lys 145 150 155 160 tac
tcg cca acc ggc aac atc gat aat gga acg tct cct caa tta atg 528Tyr
Ser Pro Thr Gly Asn Ile Asp Asn Gly Thr Ser Pro Gln Leu Met 165 170
175 ggc tgg acc ctc gta ccc tgt ata atg gtg aac gac ccg aat atc gat
576Gly Trp Thr Leu Val Pro Cys Ile Met Val Asn Asp Pro Asn Ile Asp
180 185 190 aaa aat act caa att aaa acc acg ccg tac tac ata ctc aaa
aaa tac 624Lys Asn Thr Gln Ile Lys Thr Thr Pro Tyr Tyr Ile Leu Lys
Lys Tyr 195 200 205 caa tac tgg cag cgc gcg gtt ggc tca aac gtc gct
ctg cgg cca cat 672Gln Tyr Trp Gln Arg Ala Val Gly Ser Asn Val Ala
Leu Arg Pro His 210 215 220 gaa aag aag tcc tac act tac gaa tgg ggc
aca gag atc gat cag aaa 720Glu Lys Lys Ser Tyr Thr Tyr Glu Trp Gly
Thr Glu Ile Asp Gln Lys 225 230 235 240 acg acc att ata aat aca tta
gga ttc caa atc aat atc gac agc gga 768Thr Thr Ile Ile Asn Thr Leu
Gly Phe Gln Ile Asn Ile Asp Ser Gly 245 250 255 atg aaa ttt gac atc
ccg gaa gtg ggg ggc ggg acc gat gaa ata aaa 816Met Lys Phe Asp Ile
Pro Glu Val Gly Gly Gly Thr Asp Glu Ile Lys 260 265 270 acg cag ctc
aac gaa gaa tta aaa ata gag tac agt cat gaa act aaa 864Thr Gln Leu
Asn Glu Glu Leu Lys Ile Glu Tyr Ser His Glu Thr Lys 275 280 285 ata
atg gaa aaa tat caa gag caa tct gaa atc gat aac ccg acc gac 912Ile
Met Glu Lys Tyr Gln Glu Gln Ser Glu Ile Asp Asn Pro Thr Asp 290 295
300 caa tca atg aac tct atc ggt ttc ctt act att acc tcc ctg gag ttg
960Gln Ser Met Asn Ser Ile Gly Phe Leu Thr Ile Thr Ser Leu Glu Leu
305 310 315 320 tac aga tat aac ggc tct gag atc cgt ata atg cag att
caa acc tca 1008Tyr Arg Tyr Asn Gly Ser Glu Ile Arg Ile Met Gln Ile
Gln Thr Ser 325 330 335 gac aat gac act tat aac gtc acc tct tac ccg
aat cat cag caa gcc 1056Asp Asn Asp Thr Tyr Asn Val Thr Ser Tyr Pro
Asn His Gln Gln Ala 340 345 350 ctg ctg ctt ctt aca aac cac agt tat
gaa gag gtg gaa gag ata acg 1104Leu Leu Leu Leu Thr Asn His Ser Tyr
Glu Glu Val Glu Glu Ile Thr 355 360 365 aac att cct aaa tcc aca cta
aag aaa tta aaa aaa tat tat ttc tga 1152Asn Ile Pro Lys Ser Thr Leu
Lys Lys Leu Lys Lys Tyr Tyr Phe 370 375 380 18383PRTArtificial
SequenceSynthetic Construct 18Met Leu Asp Thr Asn Lys Val Tyr Glu
Ile Ser Asn His Ala Asn Gly 1 5 10 15 Leu Tyr Ala Ala Thr Tyr Leu
Ser Leu Asp Asp Ser Gly Val Ser Leu 20 25 30 Met Asn Lys Asn Asp
Asp Asp Ile Asp Asp Tyr Asn Leu Lys Trp Phe 35 40 45 Leu Phe Pro
Ile Asp Asp Asp Gln Tyr Ile Ile Thr Ser Tyr Ala Ala 50 55 60 Asn
Asn Cys Lys Val Trp Asn Val Asn Asn Asp Lys Ile Asn Val Ser 65 70
75 80 Thr Tyr Ser Ser Thr Asn Ser Ile Gln Lys Trp Gln Ile Lys Ala
Asn 85 90 95 Gly Ser Ser Tyr Val Ile Gln Ser Asp Asn Gly Lys Val
Leu Thr Ala 100 105 110 Gly Thr Gly Gln Ala Leu Gly Leu Ile Arg Leu
Thr Asp Glu Ser Ser 115 120 125 Asn Asn Pro Asn Gln Gln Trp Asn Leu
Thr Ser Val Gln Thr Ile Gln 130 135 140 Leu Pro Gln Lys Pro Ile Ile
Asp Thr Lys Leu Lys Asp Tyr Pro Lys 145 150 155 160 Tyr Ser Pro Thr
Gly Asn Ile Asp Asn Gly Thr Ser Pro Gln Leu Met 165 170 175 Gly Trp
Thr Leu Val Pro Cys Ile Met Val Asn Asp Pro Asn Ile Asp 180 185 190
Lys Asn Thr Gln Ile Lys Thr Thr Pro Tyr Tyr Ile Leu Lys Lys Tyr 195
200 205 Gln Tyr Trp Gln Arg Ala Val Gly Ser Asn Val Ala Leu Arg Pro
His 210 215 220 Glu Lys Lys Ser Tyr Thr Tyr Glu Trp Gly Thr Glu Ile
Asp Gln Lys 225 230 235 240 Thr Thr Ile Ile Asn Thr Leu Gly Phe Gln
Ile Asn Ile Asp Ser Gly 245
250 255 Met Lys Phe Asp Ile Pro Glu Val Gly Gly Gly Thr Asp Glu Ile
Lys 260 265 270 Thr Gln Leu Asn Glu Glu Leu Lys Ile Glu Tyr Ser His
Glu Thr Lys 275 280 285 Ile Met Glu Lys Tyr Gln Glu Gln Ser Glu Ile
Asp Asn Pro Thr Asp 290 295 300 Gln Ser Met Asn Ser Ile Gly Phe Leu
Thr Ile Thr Ser Leu Glu Leu 305 310 315 320 Tyr Arg Tyr Asn Gly Ser
Glu Ile Arg Ile Met Gln Ile Gln Thr Ser 325 330 335 Asp Asn Asp Thr
Tyr Asn Val Thr Ser Tyr Pro Asn His Gln Gln Ala 340 345 350 Leu Leu
Leu Leu Thr Asn His Ser Tyr Glu Glu Val Glu Glu Ile Thr 355 360 365
Asn Ile Pro Lys Ser Thr Leu Lys Lys Leu Lys Lys Tyr Tyr Phe 370 375
380 191830DNABacillus thuringiensisCDS(1)..(1830)Native DNA
sequence encoding Bacillus thuringiensis Cry1Ab1 core toxin 19atg
gat aac aat ccg aac atc aat gaa tgc att cct tat aat tgt tta 48Met
Asp Asn Asn Pro Asn Ile Asn Glu Cys Ile Pro Tyr Asn Cys Leu 1 5 10
15 agt aac cct gaa gta gaa gta tta ggt gga gaa aga ata gaa act ggt
96Ser Asn Pro Glu Val Glu Val Leu Gly Gly Glu Arg Ile Glu Thr Gly
20 25 30 tac acc cca atc gat att tcc ttg tcg cta acg caa ttt ctt
ttg agt 144Tyr Thr Pro Ile Asp Ile Ser Leu Ser Leu Thr Gln Phe Leu
Leu Ser 35 40 45 gaa ttt gtt ccc ggt gct gga ttt gtg tta gga cta
gtt gat ata ata 192Glu Phe Val Pro Gly Ala Gly Phe Val Leu Gly Leu
Val Asp Ile Ile 50 55 60 tgg gga att ttt ggt ccc tct caa tgg gac
gca ttt ctt gta caa att 240Trp Gly Ile Phe Gly Pro Ser Gln Trp Asp
Ala Phe Leu Val Gln Ile 65 70 75 80 gaa cag tta att aac caa aga ata
gaa gaa ttc gct agg aac caa gcc 288Glu Gln Leu Ile Asn Gln Arg Ile
Glu Glu Phe Ala Arg Asn Gln Ala 85 90 95 att tct aga tta gaa gga
cta agc aat ctt tat caa att tac gca gaa 336Ile Ser Arg Leu Glu Gly
Leu Ser Asn Leu Tyr Gln Ile Tyr Ala Glu 100 105 110 tct ttt aga gag
tgg gaa gca gat cct act aat cca gca tta aga gaa 384Ser Phe Arg Glu
Trp Glu Ala Asp Pro Thr Asn Pro Ala Leu Arg Glu 115 120 125 gag atg
cgt att caa ttc aat gac atg aac agt gcc ctt aca acc gct 432Glu Met
Arg Ile Gln Phe Asn Asp Met Asn Ser Ala Leu Thr Thr Ala 130 135 140
att cct ctt ttt gca gtt caa aat tat caa gtt cct ctt tta tca gta
480Ile Pro Leu Phe Ala Val Gln Asn Tyr Gln Val Pro Leu Leu Ser Val
145 150 155 160 tat gtt caa gct gca aat tta cat tta tca gtt ttg aga
gat gtt tca 528Tyr Val Gln Ala Ala Asn Leu His Leu Ser Val Leu Arg
Asp Val Ser 165 170 175 gtg ttt gga caa agg tgg gga ttt gat gcc gcg
act atc aat agt cgt 576Val Phe Gly Gln Arg Trp Gly Phe Asp Ala Ala
Thr Ile Asn Ser Arg 180 185 190 tat aat gat tta act agg ctt att ggc
aac tat aca gat tat gct gta 624Tyr Asn Asp Leu Thr Arg Leu Ile Gly
Asn Tyr Thr Asp Tyr Ala Val 195 200 205 cgc tgg tac aat acg gga tta
gaa cgt gta tgg gga ccg gat tct aga 672Arg Trp Tyr Asn Thr Gly Leu
Glu Arg Val Trp Gly Pro Asp Ser Arg 210 215 220 gat tgg gta agg tat
aat caa ttt aga aga gaa tta aca cta act gta 720Asp Trp Val Arg Tyr
Asn Gln Phe Arg Arg Glu Leu Thr Leu Thr Val 225 230 235 240 tta gat
atc gtt gct ctg ttc ccg aat tat gat agt aga aga tat cca 768Leu Asp
Ile Val Ala Leu Phe Pro Asn Tyr Asp Ser Arg Arg Tyr Pro 245 250 255
att cga aca gtt tcc caa tta aca aga gaa att tat aca aac cca gta
816Ile Arg Thr Val Ser Gln Leu Thr Arg Glu Ile Tyr Thr Asn Pro Val
260 265 270 tta gaa aat ttt gat ggt agt ttt cga ggc tcg gct cag ggc
ata gaa 864Leu Glu Asn Phe Asp Gly Ser Phe Arg Gly Ser Ala Gln Gly
Ile Glu 275 280 285 aga agt att agg agt cca cat ttg atg gat ata ctt
aac agt ata acc 912Arg Ser Ile Arg Ser Pro His Leu Met Asp Ile Leu
Asn Ser Ile Thr 290 295 300 atc tat acg gat gct cat agg ggt tat tat
tat tgg tca ggg cat caa 960Ile Tyr Thr Asp Ala His Arg Gly Tyr Tyr
Tyr Trp Ser Gly His Gln 305 310 315 320 ata atg gct tct cct gta ggg
ttt tcg ggg cca gaa ttc act ttt ccg 1008Ile Met Ala Ser Pro Val Gly
Phe Ser Gly Pro Glu Phe Thr Phe Pro 325 330 335 cta tat gga act atg
gga aat gca gct cca caa caa cgt att gtt gct 1056Leu Tyr Gly Thr Met
Gly Asn Ala Ala Pro Gln Gln Arg Ile Val Ala 340 345 350 caa cta ggt
cag ggc gtg tat aga aca tta tcg tcc act tta tat aga 1104Gln Leu Gly
Gln Gly Val Tyr Arg Thr Leu Ser Ser Thr Leu Tyr Arg 355 360 365 aga
cct ttt aat ata ggg ata aat aat caa caa cta tct gtt ctt gac 1152Arg
Pro Phe Asn Ile Gly Ile Asn Asn Gln Gln Leu Ser Val Leu Asp 370 375
380 ggg aca gaa ttt gct tat gga acc tcc tca aat ttg cca tcc gct gta
1200Gly Thr Glu Phe Ala Tyr Gly Thr Ser Ser Asn Leu Pro Ser Ala Val
385 390 395 400 tac aga aaa agc gga acg gta gat tcg ctg gat gaa ata
ccg cca cag 1248Tyr Arg Lys Ser Gly Thr Val Asp Ser Leu Asp Glu Ile
Pro Pro Gln 405 410 415 aat aac aac gtg cca cct agg caa gga ttt agt
cat cga tta agc cat 1296Asn Asn Asn Val Pro Pro Arg Gln Gly Phe Ser
His Arg Leu Ser His 420 425 430 gtt tca atg ttt cgt tca ggc ttt agt
aat agt agt gta agt ata ata 1344Val Ser Met Phe Arg Ser Gly Phe Ser
Asn Ser Ser Val Ser Ile Ile 435 440 445 aga gct cct atg ttc tct tgg
ata cat cgt agt gct gaa ttt aat aat 1392Arg Ala Pro Met Phe Ser Trp
Ile His Arg Ser Ala Glu Phe Asn Asn 450 455 460 ata att cct tca tca
caa att aca caa ata cct tta aca aaa tct act 1440Ile Ile Pro Ser Ser
Gln Ile Thr Gln Ile Pro Leu Thr Lys Ser Thr 465 470 475 480 aat ctt
ggc tct gga act tct gtc gtt aaa gga cca gga ttt aca gga 1488Asn Leu
Gly Ser Gly Thr Ser Val Val Lys Gly Pro Gly Phe Thr Gly 485 490 495
gga gat att ctt cga aga act tca cct ggc cag att tca acc tta aga
1536Gly Asp Ile Leu Arg Arg Thr Ser Pro Gly Gln Ile Ser Thr Leu Arg
500 505 510 gta aat att act gca cca tta tca caa aga tat cgg gta aga
att cgc 1584Val Asn Ile Thr Ala Pro Leu Ser Gln Arg Tyr Arg Val Arg
Ile Arg 515 520 525 tac gct tct acc aca aat tta caa ttc cat aca tca
att gac gga aga 1632Tyr Ala Ser Thr Thr Asn Leu Gln Phe His Thr Ser
Ile Asp Gly Arg 530 535 540 cct att aat cag ggg aat ttt tca gca act
atg agt agt ggg agt aat 1680Pro Ile Asn Gln Gly Asn Phe Ser Ala Thr
Met Ser Ser Gly Ser Asn 545 550 555 560 tta cag tcc gga agc ttt agg
act gta ggt ttt act act ccg ttt aac 1728Leu Gln Ser Gly Ser Phe Arg
Thr Val Gly Phe Thr Thr Pro Phe Asn 565 570 575 ttt tca aat gga tca
agt gta ttt acg tta agt gct cat gtc ttc aat 1776Phe Ser Asn Gly Ser
Ser Val Phe Thr Leu Ser Ala His Val Phe Asn 580 585 590 tca ggc aat
gaa gtt tat ata gat cga att gaa ttt gtt ccg gca gaa 1824Ser Gly Asn
Glu Val Tyr Ile Asp Arg Ile Glu Phe Val Pro Ala Glu 595 600 605 gta
acc 1830Val Thr 610 20610PRTBacillus thuringiensis 20Met Asp Asn
Asn Pro Asn Ile Asn Glu Cys Ile Pro Tyr Asn Cys Leu 1 5 10 15 Ser
Asn Pro Glu Val Glu Val Leu Gly Gly Glu Arg Ile Glu Thr Gly 20 25
30 Tyr Thr Pro Ile Asp Ile Ser Leu Ser Leu Thr Gln Phe Leu Leu Ser
35 40 45 Glu Phe Val Pro Gly Ala Gly Phe Val Leu Gly Leu Val Asp
Ile Ile 50 55 60 Trp Gly Ile Phe Gly Pro Ser Gln Trp Asp Ala Phe
Leu Val Gln Ile 65 70 75 80 Glu Gln Leu Ile Asn Gln Arg Ile Glu Glu
Phe Ala Arg Asn Gln Ala 85 90 95 Ile Ser Arg Leu Glu Gly Leu Ser
Asn Leu Tyr Gln Ile Tyr Ala Glu 100 105 110 Ser Phe Arg Glu Trp Glu
Ala Asp Pro Thr Asn Pro Ala Leu Arg Glu 115 120 125 Glu Met Arg Ile
Gln Phe Asn Asp Met Asn Ser Ala Leu Thr Thr Ala 130 135 140 Ile Pro
Leu Phe Ala Val Gln Asn Tyr Gln Val Pro Leu Leu Ser Val 145 150 155
160 Tyr Val Gln Ala Ala Asn Leu His Leu Ser Val Leu Arg Asp Val Ser
165 170 175 Val Phe Gly Gln Arg Trp Gly Phe Asp Ala Ala Thr Ile Asn
Ser Arg 180 185 190 Tyr Asn Asp Leu Thr Arg Leu Ile Gly Asn Tyr Thr
Asp Tyr Ala Val 195 200 205 Arg Trp Tyr Asn Thr Gly Leu Glu Arg Val
Trp Gly Pro Asp Ser Arg 210 215 220 Asp Trp Val Arg Tyr Asn Gln Phe
Arg Arg Glu Leu Thr Leu Thr Val 225 230 235 240 Leu Asp Ile Val Ala
Leu Phe Pro Asn Tyr Asp Ser Arg Arg Tyr Pro 245 250 255 Ile Arg Thr
Val Ser Gln Leu Thr Arg Glu Ile Tyr Thr Asn Pro Val 260 265 270 Leu
Glu Asn Phe Asp Gly Ser Phe Arg Gly Ser Ala Gln Gly Ile Glu 275 280
285 Arg Ser Ile Arg Ser Pro His Leu Met Asp Ile Leu Asn Ser Ile Thr
290 295 300 Ile Tyr Thr Asp Ala His Arg Gly Tyr Tyr Tyr Trp Ser Gly
His Gln 305 310 315 320 Ile Met Ala Ser Pro Val Gly Phe Ser Gly Pro
Glu Phe Thr Phe Pro 325 330 335 Leu Tyr Gly Thr Met Gly Asn Ala Ala
Pro Gln Gln Arg Ile Val Ala 340 345 350 Gln Leu Gly Gln Gly Val Tyr
Arg Thr Leu Ser Ser Thr Leu Tyr Arg 355 360 365 Arg Pro Phe Asn Ile
Gly Ile Asn Asn Gln Gln Leu Ser Val Leu Asp 370 375 380 Gly Thr Glu
Phe Ala Tyr Gly Thr Ser Ser Asn Leu Pro Ser Ala Val 385 390 395 400
Tyr Arg Lys Ser Gly Thr Val Asp Ser Leu Asp Glu Ile Pro Pro Gln 405
410 415 Asn Asn Asn Val Pro Pro Arg Gln Gly Phe Ser His Arg Leu Ser
His 420 425 430 Val Ser Met Phe Arg Ser Gly Phe Ser Asn Ser Ser Val
Ser Ile Ile 435 440 445 Arg Ala Pro Met Phe Ser Trp Ile His Arg Ser
Ala Glu Phe Asn Asn 450 455 460 Ile Ile Pro Ser Ser Gln Ile Thr Gln
Ile Pro Leu Thr Lys Ser Thr 465 470 475 480 Asn Leu Gly Ser Gly Thr
Ser Val Val Lys Gly Pro Gly Phe Thr Gly 485 490 495 Gly Asp Ile Leu
Arg Arg Thr Ser Pro Gly Gln Ile Ser Thr Leu Arg 500 505 510 Val Asn
Ile Thr Ala Pro Leu Ser Gln Arg Tyr Arg Val Arg Ile Arg 515 520 525
Tyr Ala Ser Thr Thr Asn Leu Gln Phe His Thr Ser Ile Asp Gly Arg 530
535 540 Pro Ile Asn Gln Gly Asn Phe Ser Ala Thr Met Ser Ser Gly Ser
Asn 545 550 555 560 Leu Gln Ser Gly Ser Phe Arg Thr Val Gly Phe Thr
Thr Pro Phe Asn 565 570 575 Phe Ser Asn Gly Ser Ser Val Phe Thr Leu
Ser Ala His Val Phe Asn 580 585 590 Ser Gly Asn Glu Val Tyr Ile Asp
Arg Ile Glu Phe Val Pro Ala Glu 595 600 605 Val Thr 610
211830DNAArtificial SequenceSynthetic DNA sequence encoding
Bacillus thuringiensis Cry1Ab1 core toxin using codons optimized
for maize and Table 1 sequences are maintained 21atg gat aac aac
ccg aac atc aat gag tgc atc ccg tat aac tgt ctc 48Met Asp Asn Asn
Pro Asn Ile Asn Glu Cys Ile Pro Tyr Asn Cys Leu 1 5 10 15 agt aac
cct gaa gtg gag gtc tta ggt ggc gaa cgc atc gaa act ggt 96Ser Asn
Pro Glu Val Glu Val Leu Gly Gly Glu Arg Ile Glu Thr Gly 20 25 30
tac acc cca atc gac att agc ttg tcg ttg acg cag ttc ctt ttg tcc
144Tyr Thr Pro Ile Asp Ile Ser Leu Ser Leu Thr Gln Phe Leu Leu Ser
35 40 45 gag ttc gtg ccc ggt gcg ggt ttc gtg ctg ggg cta gtt gat
ata atc 192Glu Phe Val Pro Gly Ala Gly Phe Val Leu Gly Leu Val Asp
Ile Ile 50 55 60 tgg gga atc ttt ggt ccc tct cag tgg gac gcc ttt
ctt gtg caa att 240Trp Gly Ile Phe Gly Pro Ser Gln Trp Asp Ala Phe
Leu Val Gln Ile 65 70 75 80 gag cag cta att aac caa aga ata gaa gag
ttc gcg agg aac caa gcc 288Glu Gln Leu Ile Asn Gln Arg Ile Glu Glu
Phe Ala Arg Asn Gln Ala 85 90 95 att tcc aga ctg gag gga cta agc
aac ctt tat caa atc tac gcg gag 336Ile Ser Arg Leu Glu Gly Leu Ser
Asn Leu Tyr Gln Ile Tyr Ala Glu 100 105 110 tct ttt agg gag tgg gag
gca gat cct acg aac ccg gca ctg cgc gaa 384Ser Phe Arg Glu Trp Glu
Ala Asp Pro Thr Asn Pro Ala Leu Arg Glu 115 120 125 gag atg cgt att
cag ttc aac gac atg aac agt gcc ctt aca acc gct 432Glu Met Arg Ile
Gln Phe Asn Asp Met Asn Ser Ala Leu Thr Thr Ala 130 135 140 att ccc
ctt ttc gca gtt caa aat tac caa gtt ccc ctt ctc tca gtg 480Ile Pro
Leu Phe Ala Val Gln Asn Tyr Gln Val Pro Leu Leu Ser Val 145 150 155
160 tac gtt caa gcc gca aat tta cac cta agc gtt ctc cgc gat gtg tca
528Tyr Val Gln Ala Ala Asn Leu His Leu Ser Val Leu Arg Asp Val Ser
165 170 175 gtg ttt ggc cag agg tgg gga ttt gat gcc gcc act atc aat
agt cgt 576Val Phe Gly Gln Arg Trp Gly Phe Asp Ala Ala Thr Ile Asn
Ser Arg 180 185 190 tat aat gat ctg acg agg ctt atc ggc aac tat acc
gac tat gct gtc 624Tyr Asn Asp Leu Thr Arg Leu Ile Gly Asn Tyr Thr
Asp Tyr Ala Val 195 200 205 cgc tgg tac aat acg gga tta gag cgg gtc
tgg ggt ccg gat tcc cga 672Arg Trp Tyr Asn Thr Gly Leu Glu Arg Val
Trp Gly Pro Asp Ser Arg 210 215 220 gac tgg gtg cgc tac aat caa ttc
cgc cgc gaa tta acc ctc act gtc 720Asp Trp Val Arg Tyr Asn Gln Phe
Arg Arg Glu Leu Thr Leu Thr Val 225 230 235 240 ctc gac atc gtg gcg
ctg ttc ccg aac tac gac agt agg aga tac cca 768Leu Asp Ile Val Ala
Leu Phe Pro Asn Tyr Asp Ser Arg Arg Tyr Pro 245 250 255 atc cgc aca
gtt tcc caa tta acg cgg gaa att tac acc aac cca gtc
816Ile Arg Thr Val Ser Gln Leu Thr Arg Glu Ile Tyr Thr Asn Pro Val
260 265 270 ctg gag aat ttt gac ggg agc ttc cga ggc tcg gct caa ggc
ata gaa 864Leu Glu Asn Phe Asp Gly Ser Phe Arg Gly Ser Ala Gln Gly
Ile Glu 275 280 285 cgc agc att agg tcg cca cac ttg atg gat atc ctt
aac agc atc acc 912Arg Ser Ile Arg Ser Pro His Leu Met Asp Ile Leu
Asn Ser Ile Thr 290 295 300 atc tac acg gat gcc cat agg ggt tac tac
tac tgg tcg ggg cat caa 960Ile Tyr Thr Asp Ala His Arg Gly Tyr Tyr
Tyr Trp Ser Gly His Gln 305 310 315 320 ata atg gct tct cct gtc ggg
ttt tcg ggg cca gag ttc acc ttc ccg 1008Ile Met Ala Ser Pro Val Gly
Phe Ser Gly Pro Glu Phe Thr Phe Pro 325 330 335 ctc tac ggc act atg
gga aat gcc gcg cca caa caa cgt atc gtc gct 1056Leu Tyr Gly Thr Met
Gly Asn Ala Ala Pro Gln Gln Arg Ile Val Ala 340 345 350 caa cta ggt
caa ggc gtg tac cgg aca ctg tcg tcc act ctc tat cgg 1104Gln Leu Gly
Gln Gly Val Tyr Arg Thr Leu Ser Ser Thr Leu Tyr Arg 355 360 365 cgg
cct ttc aat ata ggg ata aat aat caa cag ttg tct gtg ctg gac 1152Arg
Pro Phe Asn Ile Gly Ile Asn Asn Gln Gln Leu Ser Val Leu Asp 370 375
380 ggg aca gag ttt gct tac gga acc tca agc aac ttg cca tcc gct gta
1200Gly Thr Glu Phe Ala Tyr Gly Thr Ser Ser Asn Leu Pro Ser Ala Val
385 390 395 400 tac aga aaa agc ggc acg gtg gac tcg ctg gat gaa atc
ccg ccc cag 1248Tyr Arg Lys Ser Gly Thr Val Asp Ser Leu Asp Glu Ile
Pro Pro Gln 405 410 415 aat aac aac gtg ccc cct cgg caa ggc ttc agt
cat cga ctg agc cac 1296Asn Asn Asn Val Pro Pro Arg Gln Gly Phe Ser
His Arg Leu Ser His 420 425 430 gtt agc atg ttc cgt tcg ggc ttc agc
aac tcc tcc gta agt ata ata 1344Val Ser Met Phe Arg Ser Gly Phe Ser
Asn Ser Ser Val Ser Ile Ile 435 440 445 aga gca cct atg ttc agc tgg
ata cat cgt tcc gcc gag ttt aat aat 1392Arg Ala Pro Met Phe Ser Trp
Ile His Arg Ser Ala Glu Phe Asn Asn 450 455 460 ata att ccc tcc tct
caa atc aca cag atc cct ctg aca aag tct act 1440Ile Ile Pro Ser Ser
Gln Ile Thr Gln Ile Pro Leu Thr Lys Ser Thr 465 470 475 480 aat ctt
ggc tct ggg act tct gtc gtt aag ggg cct ggc ttt acg ggc 1488Asn Leu
Gly Ser Gly Thr Ser Val Val Lys Gly Pro Gly Phe Thr Gly 485 490 495
ggc gat att ctg cgg aga act tca cct ggc cag att tcc acc ctg cgc
1536Gly Asp Ile Leu Arg Arg Thr Ser Pro Gly Gln Ile Ser Thr Leu Arg
500 505 510 gtg aat atc acc gcg cca ttg tca caa cgt tac cgc gtg cgg
att cgc 1584Val Asn Ile Thr Ala Pro Leu Ser Gln Arg Tyr Arg Val Arg
Ile Arg 515 520 525 tac gct tct acc aca aac ctc cag ttc cat aca tct
att gac ggc aga 1632Tyr Ala Ser Thr Thr Asn Leu Gln Phe His Thr Ser
Ile Asp Gly Arg 530 535 540 ccc att aat caa ggg aat ttc tcc gcc acg
atg tcg tcc ggc tcc aat 1680Pro Ile Asn Gln Gly Asn Phe Ser Ala Thr
Met Ser Ser Gly Ser Asn 545 550 555 560 ctc cag tcc gga agt ttc cgc
acc gta ggt ttt act acc ccg ttc aac 1728Leu Gln Ser Gly Ser Phe Arg
Thr Val Gly Phe Thr Thr Pro Phe Asn 565 570 575 ttt tca aac ggc tca
agt gtg ttt acg ctg tcc gct cat gtg ttc aac 1776Phe Ser Asn Gly Ser
Ser Val Phe Thr Leu Ser Ala His Val Phe Asn 580 585 590 tct ggc aat
gag gtt tat atc gac cgg att gag ttc gtc ccg gca gaa 1824Ser Gly Asn
Glu Val Tyr Ile Asp Arg Ile Glu Phe Val Pro Ala Glu 595 600 605 gtc
acc 1830Val Thr 610 22610PRTArtificial SequenceSynthetic Construct
22Met Asp Asn Asn Pro Asn Ile Asn Glu Cys Ile Pro Tyr Asn Cys Leu 1
5 10 15 Ser Asn Pro Glu Val Glu Val Leu Gly Gly Glu Arg Ile Glu Thr
Gly 20 25 30 Tyr Thr Pro Ile Asp Ile Ser Leu Ser Leu Thr Gln Phe
Leu Leu Ser 35 40 45 Glu Phe Val Pro Gly Ala Gly Phe Val Leu Gly
Leu Val Asp Ile Ile 50 55 60 Trp Gly Ile Phe Gly Pro Ser Gln Trp
Asp Ala Phe Leu Val Gln Ile 65 70 75 80 Glu Gln Leu Ile Asn Gln Arg
Ile Glu Glu Phe Ala Arg Asn Gln Ala 85 90 95 Ile Ser Arg Leu Glu
Gly Leu Ser Asn Leu Tyr Gln Ile Tyr Ala Glu 100 105 110 Ser Phe Arg
Glu Trp Glu Ala Asp Pro Thr Asn Pro Ala Leu Arg Glu 115 120 125 Glu
Met Arg Ile Gln Phe Asn Asp Met Asn Ser Ala Leu Thr Thr Ala 130 135
140 Ile Pro Leu Phe Ala Val Gln Asn Tyr Gln Val Pro Leu Leu Ser Val
145 150 155 160 Tyr Val Gln Ala Ala Asn Leu His Leu Ser Val Leu Arg
Asp Val Ser 165 170 175 Val Phe Gly Gln Arg Trp Gly Phe Asp Ala Ala
Thr Ile Asn Ser Arg 180 185 190 Tyr Asn Asp Leu Thr Arg Leu Ile Gly
Asn Tyr Thr Asp Tyr Ala Val 195 200 205 Arg Trp Tyr Asn Thr Gly Leu
Glu Arg Val Trp Gly Pro Asp Ser Arg 210 215 220 Asp Trp Val Arg Tyr
Asn Gln Phe Arg Arg Glu Leu Thr Leu Thr Val 225 230 235 240 Leu Asp
Ile Val Ala Leu Phe Pro Asn Tyr Asp Ser Arg Arg Tyr Pro 245 250 255
Ile Arg Thr Val Ser Gln Leu Thr Arg Glu Ile Tyr Thr Asn Pro Val 260
265 270 Leu Glu Asn Phe Asp Gly Ser Phe Arg Gly Ser Ala Gln Gly Ile
Glu 275 280 285 Arg Ser Ile Arg Ser Pro His Leu Met Asp Ile Leu Asn
Ser Ile Thr 290 295 300 Ile Tyr Thr Asp Ala His Arg Gly Tyr Tyr Tyr
Trp Ser Gly His Gln 305 310 315 320 Ile Met Ala Ser Pro Val Gly Phe
Ser Gly Pro Glu Phe Thr Phe Pro 325 330 335 Leu Tyr Gly Thr Met Gly
Asn Ala Ala Pro Gln Gln Arg Ile Val Ala 340 345 350 Gln Leu Gly Gln
Gly Val Tyr Arg Thr Leu Ser Ser Thr Leu Tyr Arg 355 360 365 Arg Pro
Phe Asn Ile Gly Ile Asn Asn Gln Gln Leu Ser Val Leu Asp 370 375 380
Gly Thr Glu Phe Ala Tyr Gly Thr Ser Ser Asn Leu Pro Ser Ala Val 385
390 395 400 Tyr Arg Lys Ser Gly Thr Val Asp Ser Leu Asp Glu Ile Pro
Pro Gln 405 410 415 Asn Asn Asn Val Pro Pro Arg Gln Gly Phe Ser His
Arg Leu Ser His 420 425 430 Val Ser Met Phe Arg Ser Gly Phe Ser Asn
Ser Ser Val Ser Ile Ile 435 440 445 Arg Ala Pro Met Phe Ser Trp Ile
His Arg Ser Ala Glu Phe Asn Asn 450 455 460 Ile Ile Pro Ser Ser Gln
Ile Thr Gln Ile Pro Leu Thr Lys Ser Thr 465 470 475 480 Asn Leu Gly
Ser Gly Thr Ser Val Val Lys Gly Pro Gly Phe Thr Gly 485 490 495 Gly
Asp Ile Leu Arg Arg Thr Ser Pro Gly Gln Ile Ser Thr Leu Arg 500 505
510 Val Asn Ile Thr Ala Pro Leu Ser Gln Arg Tyr Arg Val Arg Ile Arg
515 520 525 Tyr Ala Ser Thr Thr Asn Leu Gln Phe His Thr Ser Ile Asp
Gly Arg 530 535 540 Pro Ile Asn Gln Gly Asn Phe Ser Ala Thr Met Ser
Ser Gly Ser Asn 545 550 555 560 Leu Gln Ser Gly Ser Phe Arg Thr Val
Gly Phe Thr Thr Pro Phe Asn 565 570 575 Phe Ser Asn Gly Ser Ser Val
Phe Thr Leu Ser Ala His Val Phe Asn 580 585 590 Ser Gly Asn Glu Val
Tyr Ile Asp Arg Ile Glu Phe Val Pro Ala Glu 595 600 605 Val Thr 610
231830DNAArtificial SequenceSynthetic DNA sequence in accordance
with the invention encoding Bacillus thuringiensis Cry1Ab1 core
toxin using codons optimized for maize and with sequences
identified in Table 2 removed and Table 1 sequences are maintained
23atg gat aac aac ccg aac atc aat gag tgc atc ccg tat aac tgt ctc
48Met Asp Asn Asn Pro Asn Ile Asn Glu Cys Ile Pro Tyr Asn Cys Leu 1
5 10 15 agt aac cct gaa gtg gag gtc tta ggt ggc gaa cgc atc gaa act
ggt 96Ser Asn Pro Glu Val Glu Val Leu Gly Gly Glu Arg Ile Glu Thr
Gly 20 25 30 tac acc cca atc gac att agc ttg tcg ttg acg cag ttc
ctc ttg tcc 144Tyr Thr Pro Ile Asp Ile Ser Leu Ser Leu Thr Gln Phe
Leu Leu Ser 35 40 45 gag ttc gtg ccc ggt gcg ggt ttc gtg ctg ggg
cta gtt gat ata atc 192Glu Phe Val Pro Gly Ala Gly Phe Val Leu Gly
Leu Val Asp Ile Ile 50 55 60 tgg gga atc ttt ggt ccc tct cag tgg
gac gcc ttt ctt gtg caa att 240Trp Gly Ile Phe Gly Pro Ser Gln Trp
Asp Ala Phe Leu Val Gln Ile 65 70 75 80 gag cag cta att aac caa aga
ata gaa gag ttc gcg agg aac caa gcc 288Glu Gln Leu Ile Asn Gln Arg
Ile Glu Glu Phe Ala Arg Asn Gln Ala 85 90 95 att tcc aga ctg gag
gga cta agc aac ctt tat caa atc tac gcg gag 336Ile Ser Arg Leu Glu
Gly Leu Ser Asn Leu Tyr Gln Ile Tyr Ala Glu 100 105 110 tct ttt agg
gag tgg gag gca gat cct acg aac ccg gca ctg cgc gaa 384Ser Phe Arg
Glu Trp Glu Ala Asp Pro Thr Asn Pro Ala Leu Arg Glu 115 120 125 gag
atg cgt att cag ttc aac gac atg aac agt gcc ctt aca acc gct 432Glu
Met Arg Ile Gln Phe Asn Asp Met Asn Ser Ala Leu Thr Thr Ala 130 135
140 att ccc ctt ttc gca gtt caa aat tac caa gtt ccc ctt ctc tca gtg
480Ile Pro Leu Phe Ala Val Gln Asn Tyr Gln Val Pro Leu Leu Ser Val
145 150 155 160 tac gtt caa gcc gca aat tta cac cta agc gtt ctc cgc
gat gtg tca 528Tyr Val Gln Ala Ala Asn Leu His Leu Ser Val Leu Arg
Asp Val Ser 165 170 175 gtg ttc ggc cag agg tgg gga ttt gat gcc gcc
act atc aat agt cgt 576Val Phe Gly Gln Arg Trp Gly Phe Asp Ala Ala
Thr Ile Asn Ser Arg 180 185 190 tat aat gat ctg acg agg ctt atc ggc
aac tat acc gac tat gct gtc 624Tyr Asn Asp Leu Thr Arg Leu Ile Gly
Asn Tyr Thr Asp Tyr Ala Val 195 200 205 cgc tgg tac aat acg gga tta
gag cgg gtc tgg ggt ccg gat tcc cga 672Arg Trp Tyr Asn Thr Gly Leu
Glu Arg Val Trp Gly Pro Asp Ser Arg 210 215 220 gac tgg gtg cgc tac
aat caa ttc cgc cgc gaa tta acc ctc act gtc 720Asp Trp Val Arg Tyr
Asn Gln Phe Arg Arg Glu Leu Thr Leu Thr Val 225 230 235 240 ctc gac
atc gtg gcg ctg ttc ccg aac tac gac agt agg aga tac cca 768Leu Asp
Ile Val Ala Leu Phe Pro Asn Tyr Asp Ser Arg Arg Tyr Pro 245 250 255
atc cgc aca gtt tcc caa tta acg cgg gaa att tac acc aac cca gtc
816Ile Arg Thr Val Ser Gln Leu Thr Arg Glu Ile Tyr Thr Asn Pro Val
260 265 270 ctg gag aat ttt gac ggg agc ttc cga ggc tcg gct caa ggc
ata gaa 864Leu Glu Asn Phe Asp Gly Ser Phe Arg Gly Ser Ala Gln Gly
Ile Glu 275 280 285 cgc agc att agg tcg cca cac ttg atg gat atc ctt
aac agc atc acc 912Arg Ser Ile Arg Ser Pro His Leu Met Asp Ile Leu
Asn Ser Ile Thr 290 295 300 atc tac acg gat gcc cat agg ggt tac tac
tac tgg tcg ggg cat caa 960Ile Tyr Thr Asp Ala His Arg Gly Tyr Tyr
Tyr Trp Ser Gly His Gln 305 310 315 320 ata atg gct tct cct gtc ggg
ttt tcg ggg cca gag ttc acc ttc ccg 1008Ile Met Ala Ser Pro Val Gly
Phe Ser Gly Pro Glu Phe Thr Phe Pro 325 330 335 ctc tac ggc act atg
gga aat gcc gcg cca caa caa cgt atc gtc gct 1056Leu Tyr Gly Thr Met
Gly Asn Ala Ala Pro Gln Gln Arg Ile Val Ala 340 345 350 caa cta ggt
caa ggc gtg tac cgg aca ctg tcg tcc act ctc tat cgg 1104Gln Leu Gly
Gln Gly Val Tyr Arg Thr Leu Ser Ser Thr Leu Tyr Arg 355 360 365 cgg
cct ttc aat ata ggg atc aat aat caa cag ttg tct gtg ctg gac 1152Arg
Pro Phe Asn Ile Gly Ile Asn Asn Gln Gln Leu Ser Val Leu Asp 370 375
380 ggg aca gag ttt gct tac gga acc tca agc aac ttg cca tcc gct gta
1200Gly Thr Glu Phe Ala Tyr Gly Thr Ser Ser Asn Leu Pro Ser Ala Val
385 390 395 400 tac aga aaa agc ggc acg gtg gac tcg ctg gat gaa atc
ccg ccc cag 1248Tyr Arg Lys Ser Gly Thr Val Asp Ser Leu Asp Glu Ile
Pro Pro Gln 405 410 415 aat aac aac gtg ccc cct cgg caa ggc ttc agt
cat cga ctg agc cac 1296Asn Asn Asn Val Pro Pro Arg Gln Gly Phe Ser
His Arg Leu Ser His 420 425 430 gtt agc atg ttc cgt tcg ggc ttc agc
aac tcc tcc gta agt atc ata 1344Val Ser Met Phe Arg Ser Gly Phe Ser
Asn Ser Ser Val Ser Ile Ile 435 440 445 aga gca cct atg ttc agc tgg
ata cat cgt tcc gcc gag ttc aat aat 1392Arg Ala Pro Met Phe Ser Trp
Ile His Arg Ser Ala Glu Phe Asn Asn 450 455 460 ata att ccc tcc tct
caa atc aca cag atc cct ctg aca aag tct act 1440Ile Ile Pro Ser Ser
Gln Ile Thr Gln Ile Pro Leu Thr Lys Ser Thr 465 470 475 480 aat ctt
ggc tct ggg act tct gtc gtt aag ggg cct ggc ttt acg ggc 1488Asn Leu
Gly Ser Gly Thr Ser Val Val Lys Gly Pro Gly Phe Thr Gly 485 490 495
ggc gat att ctg cgg aga act tca cct ggc cag att tcc acc ctg cgc
1536Gly Asp Ile Leu Arg Arg Thr Ser Pro Gly Gln Ile Ser Thr Leu Arg
500 505 510 gtg aat atc acc gcg cca ttg tca caa cgt tac cgc gtg cgg
att cgc 1584Val Asn Ile Thr Ala Pro Leu Ser Gln Arg Tyr Arg Val Arg
Ile Arg 515 520 525 tac gct tct acc aca aac ctc cag ttc cat aca tct
att gac ggc aga 1632Tyr Ala Ser Thr Thr Asn Leu Gln Phe His Thr Ser
Ile Asp Gly Arg 530 535 540 ccc att aat caa ggg aat ttc tcc gcc acg
atg tcg tcc ggc tcc aat 1680Pro Ile Asn Gln Gly Asn Phe Ser Ala Thr
Met Ser Ser Gly Ser Asn 545 550 555 560 ctc cag tcc gga agt ttc cgc
acc gta ggt ttt act acc ccg ttc aac 1728Leu Gln Ser Gly Ser Phe Arg
Thr Val Gly Phe Thr Thr Pro Phe Asn 565 570 575 ttt tca aac ggc tca
agt gtg ttt acg ctg tcc gct cat gtg ttc aac 1776Phe Ser Asn Gly Ser
Ser Val Phe Thr Leu Ser Ala His Val Phe Asn 580 585 590 tct ggc aat
gag gtt tac atc gac cgg att gag ttc gtc ccg gca gaa 1824Ser Gly Asn
Glu Val Tyr Ile Asp Arg Ile Glu Phe Val Pro Ala Glu
595 600 605 gtc acc 1830Val Thr 610 24610PRTArtificial
SequenceSynthetic Construct 24Met Asp Asn Asn Pro Asn Ile Asn Glu
Cys Ile Pro Tyr Asn Cys Leu 1 5 10 15 Ser Asn Pro Glu Val Glu Val
Leu Gly Gly Glu Arg Ile Glu Thr Gly 20 25 30 Tyr Thr Pro Ile Asp
Ile Ser Leu Ser Leu Thr Gln Phe Leu Leu Ser 35 40 45 Glu Phe Val
Pro Gly Ala Gly Phe Val Leu Gly Leu Val Asp Ile Ile 50 55 60 Trp
Gly Ile Phe Gly Pro Ser Gln Trp Asp Ala Phe Leu Val Gln Ile 65 70
75 80 Glu Gln Leu Ile Asn Gln Arg Ile Glu Glu Phe Ala Arg Asn Gln
Ala 85 90 95 Ile Ser Arg Leu Glu Gly Leu Ser Asn Leu Tyr Gln Ile
Tyr Ala Glu 100 105 110 Ser Phe Arg Glu Trp Glu Ala Asp Pro Thr Asn
Pro Ala Leu Arg Glu 115 120 125 Glu Met Arg Ile Gln Phe Asn Asp Met
Asn Ser Ala Leu Thr Thr Ala 130 135 140 Ile Pro Leu Phe Ala Val Gln
Asn Tyr Gln Val Pro Leu Leu Ser Val 145 150 155 160 Tyr Val Gln Ala
Ala Asn Leu His Leu Ser Val Leu Arg Asp Val Ser 165 170 175 Val Phe
Gly Gln Arg Trp Gly Phe Asp Ala Ala Thr Ile Asn Ser Arg 180 185 190
Tyr Asn Asp Leu Thr Arg Leu Ile Gly Asn Tyr Thr Asp Tyr Ala Val 195
200 205 Arg Trp Tyr Asn Thr Gly Leu Glu Arg Val Trp Gly Pro Asp Ser
Arg 210 215 220 Asp Trp Val Arg Tyr Asn Gln Phe Arg Arg Glu Leu Thr
Leu Thr Val 225 230 235 240 Leu Asp Ile Val Ala Leu Phe Pro Asn Tyr
Asp Ser Arg Arg Tyr Pro 245 250 255 Ile Arg Thr Val Ser Gln Leu Thr
Arg Glu Ile Tyr Thr Asn Pro Val 260 265 270 Leu Glu Asn Phe Asp Gly
Ser Phe Arg Gly Ser Ala Gln Gly Ile Glu 275 280 285 Arg Ser Ile Arg
Ser Pro His Leu Met Asp Ile Leu Asn Ser Ile Thr 290 295 300 Ile Tyr
Thr Asp Ala His Arg Gly Tyr Tyr Tyr Trp Ser Gly His Gln 305 310 315
320 Ile Met Ala Ser Pro Val Gly Phe Ser Gly Pro Glu Phe Thr Phe Pro
325 330 335 Leu Tyr Gly Thr Met Gly Asn Ala Ala Pro Gln Gln Arg Ile
Val Ala 340 345 350 Gln Leu Gly Gln Gly Val Tyr Arg Thr Leu Ser Ser
Thr Leu Tyr Arg 355 360 365 Arg Pro Phe Asn Ile Gly Ile Asn Asn Gln
Gln Leu Ser Val Leu Asp 370 375 380 Gly Thr Glu Phe Ala Tyr Gly Thr
Ser Ser Asn Leu Pro Ser Ala Val 385 390 395 400 Tyr Arg Lys Ser Gly
Thr Val Asp Ser Leu Asp Glu Ile Pro Pro Gln 405 410 415 Asn Asn Asn
Val Pro Pro Arg Gln Gly Phe Ser His Arg Leu Ser His 420 425 430 Val
Ser Met Phe Arg Ser Gly Phe Ser Asn Ser Ser Val Ser Ile Ile 435 440
445 Arg Ala Pro Met Phe Ser Trp Ile His Arg Ser Ala Glu Phe Asn Asn
450 455 460 Ile Ile Pro Ser Ser Gln Ile Thr Gln Ile Pro Leu Thr Lys
Ser Thr 465 470 475 480 Asn Leu Gly Ser Gly Thr Ser Val Val Lys Gly
Pro Gly Phe Thr Gly 485 490 495 Gly Asp Ile Leu Arg Arg Thr Ser Pro
Gly Gln Ile Ser Thr Leu Arg 500 505 510 Val Asn Ile Thr Ala Pro Leu
Ser Gln Arg Tyr Arg Val Arg Ile Arg 515 520 525 Tyr Ala Ser Thr Thr
Asn Leu Gln Phe His Thr Ser Ile Asp Gly Arg 530 535 540 Pro Ile Asn
Gln Gly Asn Phe Ser Ala Thr Met Ser Ser Gly Ser Asn 545 550 555 560
Leu Gln Ser Gly Ser Phe Arg Thr Val Gly Phe Thr Thr Pro Phe Asn 565
570 575 Phe Ser Asn Gly Ser Ser Val Phe Thr Leu Ser Ala His Val Phe
Asn 580 585 590 Ser Gly Asn Glu Val Tyr Ile Asp Arg Ile Glu Phe Val
Pro Ala Glu 595 600 605 Val Thr 610 251866DNABacillus
thuringiensisCDS(1)..(1866)Native DNA sequence encoding Bacillus
thuringiensis Cry1Ca core toxin 25atg gat aac aat ccg aac atc aat
gaa tgc atc ccg tac aac tgc ctg 48Met Asp Asn Asn Pro Asn Ile Asn
Glu Cys Ile Pro Tyr Asn Cys Leu 1 5 10 15 agc aac ccg gaa gaa gtg
ctg ttg gat gga gaa cgg ata tca act ggt 96Ser Asn Pro Glu Glu Val
Leu Leu Asp Gly Glu Arg Ile Ser Thr Gly 20 25 30 aat tca tca att
gat att tct ctg tca ctt gtt cag ttt ctg gta tct 144Asn Ser Ser Ile
Asp Ile Ser Leu Ser Leu Val Gln Phe Leu Val Ser 35 40 45 aac ttc
gtc cca ggc gga gga ttc ctg gtt gga tta ata gat ttt gta 192Asn Phe
Val Pro Gly Gly Gly Phe Leu Val Gly Leu Ile Asp Phe Val 50 55 60
tgg gga ata gtt ggc cct tct caa tgg gat gca ttt cta gta caa att
240Trp Gly Ile Val Gly Pro Ser Gln Trp Asp Ala Phe Leu Val Gln Ile
65 70 75 80 gaa caa tta att aat gaa aga ata gct gaa ttt gct agg aat
gct gct 288Glu Gln Leu Ile Asn Glu Arg Ile Ala Glu Phe Ala Arg Asn
Ala Ala 85 90 95 att gct aat tta gaa gga tta gga aac aat ttc aat
ata tat gtg gaa 336Ile Ala Asn Leu Glu Gly Leu Gly Asn Asn Phe Asn
Ile Tyr Val Glu 100 105 110 gca ttt aaa gaa tgg gaa gaa gat cct aag
aat cca gca acc agg acc 384Ala Phe Lys Glu Trp Glu Glu Asp Pro Lys
Asn Pro Ala Thr Arg Thr 115 120 125 aga gta att gat cgc ttt cgt ata
ctt gat ggg cta ctt gaa agg gac 432Arg Val Ile Asp Arg Phe Arg Ile
Leu Asp Gly Leu Leu Glu Arg Asp 130 135 140 att cct tcg ttt cga att
tct gga ttt gaa gta ccc ctt tta tcc gtt 480Ile Pro Ser Phe Arg Ile
Ser Gly Phe Glu Val Pro Leu Leu Ser Val 145 150 155 160 tat gct caa
gcg gcc aat ctg cat cta gct ata tta aga gat tct gta 528Tyr Ala Gln
Ala Ala Asn Leu His Leu Ala Ile Leu Arg Asp Ser Val 165 170 175 att
ttt gga gaa aga tgg gga ttg aca acg ata aat gtc aat gaa aac 576Ile
Phe Gly Glu Arg Trp Gly Leu Thr Thr Ile Asn Val Asn Glu Asn 180 185
190 tat aat aga cta att agg cat att gat gaa tat gct gat cac tgt gca
624Tyr Asn Arg Leu Ile Arg His Ile Asp Glu Tyr Ala Asp His Cys Ala
195 200 205 aat acg tat aat cgg gga tta aat aat tta ccg aaa tct acg
tat caa 672Asn Thr Tyr Asn Arg Gly Leu Asn Asn Leu Pro Lys Ser Thr
Tyr Gln 210 215 220 gat tgg ata aca tat aat cga ttg cgg aga gac tta
aca ttg act gta 720Asp Trp Ile Thr Tyr Asn Arg Leu Arg Arg Asp Leu
Thr Leu Thr Val 225 230 235 240 tta gat atc gcc gct ttc ttt cca aac
tat gac aat agg aga tat cca 768Leu Asp Ile Ala Ala Phe Phe Pro Asn
Tyr Asp Asn Arg Arg Tyr Pro 245 250 255 att cag cca gtt ggt caa cta
aca agg gaa gtt tat acg gac cca tta 816Ile Gln Pro Val Gly Gln Leu
Thr Arg Glu Val Tyr Thr Asp Pro Leu 260 265 270 att aat ttt aat cca
cag tta cag tct gta gct caa tta cct act ttt 864Ile Asn Phe Asn Pro
Gln Leu Gln Ser Val Ala Gln Leu Pro Thr Phe 275 280 285 aac gtt atg
gag aac agc gca att aga aat cct cat tta ttt gat ata 912Asn Val Met
Glu Asn Ser Ala Ile Arg Asn Pro His Leu Phe Asp Ile 290 295 300 ttg
aat aat ctt aca atc ttt acg gat tgg ttt agt gtt gga cgc aat 960Leu
Asn Asn Leu Thr Ile Phe Thr Asp Trp Phe Ser Val Gly Arg Asn 305 310
315 320 ttt tat tgg gga gga cat cga gta ata tct agc ctt ata gga ggt
ggt 1008Phe Tyr Trp Gly Gly His Arg Val Ile Ser Ser Leu Ile Gly Gly
Gly 325 330 335 aac ata aca tct cct ata tat gga aga gag gcg aac cag
gag cct cca 1056Asn Ile Thr Ser Pro Ile Tyr Gly Arg Glu Ala Asn Gln
Glu Pro Pro 340 345 350 aga tcc ttt act ttt aat gga ccg gta ttt agg
act tta tca aat cct 1104Arg Ser Phe Thr Phe Asn Gly Pro Val Phe Arg
Thr Leu Ser Asn Pro 355 360 365 act tta cga tta tta cag caa cct tgg
cca gcg cca cca ttt aat tta 1152Thr Leu Arg Leu Leu Gln Gln Pro Trp
Pro Ala Pro Pro Phe Asn Leu 370 375 380 cgt ggt gtt gaa gga gta gaa
ttt tct aca cct aca aat agc ttt acg 1200Arg Gly Val Glu Gly Val Glu
Phe Ser Thr Pro Thr Asn Ser Phe Thr 385 390 395 400 tat cga gga aga
ggt acg gtt gat tct tta act gaa ttg ccg cct gag 1248Tyr Arg Gly Arg
Gly Thr Val Asp Ser Leu Thr Glu Leu Pro Pro Glu 405 410 415 gat aat
agt gtg cca cct cgc gaa gga tat agt cat cgt tta tgt cat 1296Asp Asn
Ser Val Pro Pro Arg Glu Gly Tyr Ser His Arg Leu Cys His 420 425 430
gca act ttt gtt caa aga tct gga aca cct ttt tta aca act ggt gta
1344Ala Thr Phe Val Gln Arg Ser Gly Thr Pro Phe Leu Thr Thr Gly Val
435 440 445 gta ttt tct tgg acg cat cgt agt gca act ctt aca aat aca
att gat 1392Val Phe Ser Trp Thr His Arg Ser Ala Thr Leu Thr Asn Thr
Ile Asp 450 455 460 cca gag aga att aat caa ata cct tta gtg aaa gga
ttt aga gtt tgg 1440Pro Glu Arg Ile Asn Gln Ile Pro Leu Val Lys Gly
Phe Arg Val Trp 465 470 475 480 ggg ggc acc tct gtc att aca gga cca
gga ttt aca gga ggg gat atc 1488Gly Gly Thr Ser Val Ile Thr Gly Pro
Gly Phe Thr Gly Gly Asp Ile 485 490 495 ctt cga aga aat acc ttt ggt
gat ttt gta tct cta caa gtc aat att 1536Leu Arg Arg Asn Thr Phe Gly
Asp Phe Val Ser Leu Gln Val Asn Ile 500 505 510 aat tca cca att acc
caa aga tac cgt tta aga ttt cgt tac gct tcc 1584Asn Ser Pro Ile Thr
Gln Arg Tyr Arg Leu Arg Phe Arg Tyr Ala Ser 515 520 525 agt agg gat
gca cga gtt ata gta tta aca gga gcg gca tcc aca gga 1632Ser Arg Asp
Ala Arg Val Ile Val Leu Thr Gly Ala Ala Ser Thr Gly 530 535 540 gtg
gga ggc caa gtt agt gta aat atg cct ctt cag aaa act atg gaa 1680Val
Gly Gly Gln Val Ser Val Asn Met Pro Leu Gln Lys Thr Met Glu 545 550
555 560 ata ggg gag aac tta aca tct aga aca ttt aga tat acc gat ttt
agt 1728Ile Gly Glu Asn Leu Thr Ser Arg Thr Phe Arg Tyr Thr Asp Phe
Ser 565 570 575 aat cct ttt tca ttt aga gct aat cca gat ata att ggg
ata agt gaa 1776Asn Pro Phe Ser Phe Arg Ala Asn Pro Asp Ile Ile Gly
Ile Ser Glu 580 585 590 caa cct cta ttt ggt gca ggt tct att agt agc
ggt gaa ctt tat ata 1824Gln Pro Leu Phe Gly Ala Gly Ser Ile Ser Ser
Gly Glu Leu Tyr Ile 595 600 605 gat aaa att gaa att att cta gca gat
gca aca ttt gaa taa 1866Asp Lys Ile Glu Ile Ile Leu Ala Asp Ala Thr
Phe Glu 610 615 620 26621PRTBacillus thuringiensis 26Met Asp Asn
Asn Pro Asn Ile Asn Glu Cys Ile Pro Tyr Asn Cys Leu 1 5 10 15 Ser
Asn Pro Glu Glu Val Leu Leu Asp Gly Glu Arg Ile Ser Thr Gly 20 25
30 Asn Ser Ser Ile Asp Ile Ser Leu Ser Leu Val Gln Phe Leu Val Ser
35 40 45 Asn Phe Val Pro Gly Gly Gly Phe Leu Val Gly Leu Ile Asp
Phe Val 50 55 60 Trp Gly Ile Val Gly Pro Ser Gln Trp Asp Ala Phe
Leu Val Gln Ile 65 70 75 80 Glu Gln Leu Ile Asn Glu Arg Ile Ala Glu
Phe Ala Arg Asn Ala Ala 85 90 95 Ile Ala Asn Leu Glu Gly Leu Gly
Asn Asn Phe Asn Ile Tyr Val Glu 100 105 110 Ala Phe Lys Glu Trp Glu
Glu Asp Pro Lys Asn Pro Ala Thr Arg Thr 115 120 125 Arg Val Ile Asp
Arg Phe Arg Ile Leu Asp Gly Leu Leu Glu Arg Asp 130 135 140 Ile Pro
Ser Phe Arg Ile Ser Gly Phe Glu Val Pro Leu Leu Ser Val 145 150 155
160 Tyr Ala Gln Ala Ala Asn Leu His Leu Ala Ile Leu Arg Asp Ser Val
165 170 175 Ile Phe Gly Glu Arg Trp Gly Leu Thr Thr Ile Asn Val Asn
Glu Asn 180 185 190 Tyr Asn Arg Leu Ile Arg His Ile Asp Glu Tyr Ala
Asp His Cys Ala 195 200 205 Asn Thr Tyr Asn Arg Gly Leu Asn Asn Leu
Pro Lys Ser Thr Tyr Gln 210 215 220 Asp Trp Ile Thr Tyr Asn Arg Leu
Arg Arg Asp Leu Thr Leu Thr Val 225 230 235 240 Leu Asp Ile Ala Ala
Phe Phe Pro Asn Tyr Asp Asn Arg Arg Tyr Pro 245 250 255 Ile Gln Pro
Val Gly Gln Leu Thr Arg Glu Val Tyr Thr Asp Pro Leu 260 265 270 Ile
Asn Phe Asn Pro Gln Leu Gln Ser Val Ala Gln Leu Pro Thr Phe 275 280
285 Asn Val Met Glu Asn Ser Ala Ile Arg Asn Pro His Leu Phe Asp Ile
290 295 300 Leu Asn Asn Leu Thr Ile Phe Thr Asp Trp Phe Ser Val Gly
Arg Asn 305 310 315 320 Phe Tyr Trp Gly Gly His Arg Val Ile Ser Ser
Leu Ile Gly Gly Gly 325 330 335 Asn Ile Thr Ser Pro Ile Tyr Gly Arg
Glu Ala Asn Gln Glu Pro Pro 340 345 350 Arg Ser Phe Thr Phe Asn Gly
Pro Val Phe Arg Thr Leu Ser Asn Pro 355 360 365 Thr Leu Arg Leu Leu
Gln Gln Pro Trp Pro Ala Pro Pro Phe Asn Leu 370 375 380 Arg Gly Val
Glu Gly Val Glu Phe Ser Thr Pro Thr Asn Ser Phe Thr 385 390 395 400
Tyr Arg Gly Arg Gly Thr Val Asp Ser Leu Thr Glu Leu Pro Pro Glu 405
410 415 Asp Asn Ser Val Pro Pro Arg Glu Gly Tyr Ser His Arg Leu Cys
His 420 425 430 Ala Thr Phe Val Gln Arg Ser Gly Thr Pro Phe Leu Thr
Thr Gly Val 435 440 445 Val Phe Ser Trp Thr His Arg Ser Ala Thr Leu
Thr Asn Thr Ile Asp 450 455 460 Pro Glu Arg Ile Asn Gln Ile Pro Leu
Val Lys Gly Phe Arg Val Trp 465 470 475 480 Gly Gly Thr Ser Val Ile
Thr Gly Pro Gly Phe Thr Gly Gly Asp Ile 485 490 495 Leu Arg Arg Asn
Thr Phe Gly Asp Phe Val Ser Leu Gln Val Asn Ile 500 505 510 Asn Ser
Pro Ile Thr Gln Arg Tyr Arg Leu Arg Phe Arg Tyr Ala Ser 515 520 525
Ser Arg Asp Ala Arg Val Ile Val Leu Thr Gly Ala Ala Ser Thr Gly 530
535 540 Val Gly Gly Gln Val Ser Val Asn Met Pro Leu Gln Lys Thr Met
Glu 545 550 555 560 Ile Gly Glu Asn Leu Thr Ser Arg Thr Phe Arg
Tyr Thr Asp Phe Ser 565 570 575 Asn Pro Phe Ser Phe Arg Ala Asn Pro
Asp Ile Ile Gly Ile Ser Glu 580 585 590 Gln Pro Leu Phe Gly Ala Gly
Ser Ile Ser Ser Gly Glu Leu Tyr Ile 595 600 605 Asp Lys Ile Glu Ile
Ile Leu Ala Asp Ala Thr Phe Glu 610 615 620 271866DNAArtificial
SequenceSynthetic DNA sequence encoding Bacillus thuringiensis
Cry1Ca core toxin using codons optimized for maize and Table 1
sequences are maintained 27atg gat aac aat ccg aac atc aat gag tgc
atc ccg tac aac tgc ctg 48Met Asp Asn Asn Pro Asn Ile Asn Glu Cys
Ile Pro Tyr Asn Cys Leu 1 5 10 15 agc aac ccg gaa gaa gtg ctg ttg
gat gga gaa cgg ata tca act ggc 96Ser Asn Pro Glu Glu Val Leu Leu
Asp Gly Glu Arg Ile Ser Thr Gly 20 25 30 aat tca tcc att gat att
tct ctg tca ctt gtt cag ttt ctg gtg tct 144Asn Ser Ser Ile Asp Ile
Ser Leu Ser Leu Val Gln Phe Leu Val Ser 35 40 45 aac ttc gtc ccc
ggc gga gga ttc ctg gtt gga tta ata gat ttt gta 192Asn Phe Val Pro
Gly Gly Gly Phe Leu Val Gly Leu Ile Asp Phe Val 50 55 60 tgg gga
ata gtt ggc cct tct caa tgg gac gca ttt cta gta caa att 240Trp Gly
Ile Val Gly Pro Ser Gln Trp Asp Ala Phe Leu Val Gln Ile 65 70 75 80
gaa caa tta att aat gaa aga ata gct gaa ttt gct agg aac gct gct
288Glu Gln Leu Ile Asn Glu Arg Ile Ala Glu Phe Ala Arg Asn Ala Ala
85 90 95 att gct aat tta gaa gga tta gga aac aat ttc aat ata tat
gtg gaa 336Ile Ala Asn Leu Glu Gly Leu Gly Asn Asn Phe Asn Ile Tyr
Val Glu 100 105 110 gca ttt aag gaa tgg gaa gaa gat cct aag aat cca
gca acg agg acc 384Ala Phe Lys Glu Trp Glu Glu Asp Pro Lys Asn Pro
Ala Thr Arg Thr 115 120 125 aga gta att gat cgc ttt cgt ata ctt gat
ggg cta ctt gaa agg gac 432Arg Val Ile Asp Arg Phe Arg Ile Leu Asp
Gly Leu Leu Glu Arg Asp 130 135 140 att cct tcg ttt cga att tct gga
ttt gaa gta ccc ctt tta tcc gtt 480Ile Pro Ser Phe Arg Ile Ser Gly
Phe Glu Val Pro Leu Leu Ser Val 145 150 155 160 tat gct caa gcg gcc
aat ctg cat cta gct ata tta aga gat tct gta 528Tyr Ala Gln Ala Ala
Asn Leu His Leu Ala Ile Leu Arg Asp Ser Val 165 170 175 att ttt gga
gaa aga tgg gga ttg aca acg ata aat gtc aat gaa aac 576Ile Phe Gly
Glu Arg Trp Gly Leu Thr Thr Ile Asn Val Asn Glu Asn 180 185 190 tat
aat aga cta att agg cat att gat gaa tat gct gat cac tgt gca 624Tyr
Asn Arg Leu Ile Arg His Ile Asp Glu Tyr Ala Asp His Cys Ala 195 200
205 aat acg tat aat cgg gga tta aat aat tta ccg aaa tct acg tat caa
672Asn Thr Tyr Asn Arg Gly Leu Asn Asn Leu Pro Lys Ser Thr Tyr Gln
210 215 220 gat tgg ata aca tat aat cga ttg cgg aga gac tta aca ttg
act gta 720Asp Trp Ile Thr Tyr Asn Arg Leu Arg Arg Asp Leu Thr Leu
Thr Val 225 230 235 240 tta gat atc gcc gct ttc ttt cca aac tat gac
aat agg aga tat cca 768Leu Asp Ile Ala Ala Phe Phe Pro Asn Tyr Asp
Asn Arg Arg Tyr Pro 245 250 255 att cag cca gtt ggt caa cta aca agg
gaa gtt tat acg gac cca tta 816Ile Gln Pro Val Gly Gln Leu Thr Arg
Glu Val Tyr Thr Asp Pro Leu 260 265 270 att aat ttt aat cca cag tta
cag tct gta gct caa tta cct act ttt 864Ile Asn Phe Asn Pro Gln Leu
Gln Ser Val Ala Gln Leu Pro Thr Phe 275 280 285 aac gtt atg gag aac
agc gca att aga aat cct cat tta ttt gat ata 912Asn Val Met Glu Asn
Ser Ala Ile Arg Asn Pro His Leu Phe Asp Ile 290 295 300 ttg aat aat
ctt aca atc ttt acg gat tgg ttt agt gtt gga cgc aat 960Leu Asn Asn
Leu Thr Ile Phe Thr Asp Trp Phe Ser Val Gly Arg Asn 305 310 315 320
ttt tat tgg gga gga cat cga gta ata tct agc ctt ata gga ggt ggg
1008Phe Tyr Trp Gly Gly His Arg Val Ile Ser Ser Leu Ile Gly Gly Gly
325 330 335 aac atc aca tcg cct ata tat gga aga gag gcg aac caa gag
cct cca 1056Asn Ile Thr Ser Pro Ile Tyr Gly Arg Glu Ala Asn Gln Glu
Pro Pro 340 345 350 aga tcc ttt act ttt aat gga ccc gtg ttt agg act
tta tca aat cct 1104Arg Ser Phe Thr Phe Asn Gly Pro Val Phe Arg Thr
Leu Ser Asn Pro 355 360 365 act tta cga tta tta cag caa cct tgg cca
gcg cca cca ttt aat tta 1152Thr Leu Arg Leu Leu Gln Gln Pro Trp Pro
Ala Pro Pro Phe Asn Leu 370 375 380 cgt ggt gtt gaa gga gta gaa ttt
tct aca cct aca aat agc ttt acg 1200Arg Gly Val Glu Gly Val Glu Phe
Ser Thr Pro Thr Asn Ser Phe Thr 385 390 395 400 tat cga gga aga ggg
acg gtt gat tct tta act gaa ttg ccg cct gag 1248Tyr Arg Gly Arg Gly
Thr Val Asp Ser Leu Thr Glu Leu Pro Pro Glu 405 410 415 gat aat agt
gtg cca cct cgc gaa gga tat agt cat cgt tta tgt cat 1296Asp Asn Ser
Val Pro Pro Arg Glu Gly Tyr Ser His Arg Leu Cys His 420 425 430 gca
act ttt gtt caa aga tcg gga aca cct ttt tta aca act ggt gta 1344Ala
Thr Phe Val Gln Arg Ser Gly Thr Pro Phe Leu Thr Thr Gly Val 435 440
445 gta ttt tct tgg acg cat cgt agt gca act ctt aca aat aca atc gac
1392Val Phe Ser Trp Thr His Arg Ser Ala Thr Leu Thr Asn Thr Ile Asp
450 455 460 cca gag aga att aat caa ata cct tta gtg aag gga ttt aga
gtt tgg 1440Pro Glu Arg Ile Asn Gln Ile Pro Leu Val Lys Gly Phe Arg
Val Trp 465 470 475 480 ggg ggc acc tct gtc att acc gga ccc gga ttt
acc gga ggg gat atc 1488Gly Gly Thr Ser Val Ile Thr Gly Pro Gly Phe
Thr Gly Gly Asp Ile 485 490 495 ctt cga aga aat acc ttt ggt gat ttt
gta tct cta caa gtc aac att 1536Leu Arg Arg Asn Thr Phe Gly Asp Phe
Val Ser Leu Gln Val Asn Ile 500 505 510 aat tca cca att acc caa aga
tac cgt tta aga ttt cgt tac gct tcc 1584Asn Ser Pro Ile Thr Gln Arg
Tyr Arg Leu Arg Phe Arg Tyr Ala Ser 515 520 525 agt agg gat gca cga
gtt ata gta tta acg gga gcg gca tcc acc gga 1632Ser Arg Asp Ala Arg
Val Ile Val Leu Thr Gly Ala Ala Ser Thr Gly 530 535 540 gtg gga ggc
caa gtt agt gta aat atg cct ctt cag aaa act atg gaa 1680Val Gly Gly
Gln Val Ser Val Asn Met Pro Leu Gln Lys Thr Met Glu 545 550 555 560
ata ggg gag aac tta aca tcc aga aca ttt aga tat acc gat ttt agt
1728Ile Gly Glu Asn Leu Thr Ser Arg Thr Phe Arg Tyr Thr Asp Phe Ser
565 570 575 aat cct ttt tca ttt aga gct aat cca gat ata att ggg ata
agt gaa 1776Asn Pro Phe Ser Phe Arg Ala Asn Pro Asp Ile Ile Gly Ile
Ser Glu 580 585 590 caa cct cta ttt ggg gcg ggt tct att agt agc ggt
gaa ctt tat ata 1824Gln Pro Leu Phe Gly Ala Gly Ser Ile Ser Ser Gly
Glu Leu Tyr Ile 595 600 605 gat aaa att gaa att att cta gca gat gca
aca ttt gaa tga 1866Asp Lys Ile Glu Ile Ile Leu Ala Asp Ala Thr Phe
Glu 610 615 620 28621PRTArtificial SequenceSynthetic Construct
28Met Asp Asn Asn Pro Asn Ile Asn Glu Cys Ile Pro Tyr Asn Cys Leu 1
5 10 15 Ser Asn Pro Glu Glu Val Leu Leu Asp Gly Glu Arg Ile Ser Thr
Gly 20 25 30 Asn Ser Ser Ile Asp Ile Ser Leu Ser Leu Val Gln Phe
Leu Val Ser 35 40 45 Asn Phe Val Pro Gly Gly Gly Phe Leu Val Gly
Leu Ile Asp Phe Val 50 55 60 Trp Gly Ile Val Gly Pro Ser Gln Trp
Asp Ala Phe Leu Val Gln Ile 65 70 75 80 Glu Gln Leu Ile Asn Glu Arg
Ile Ala Glu Phe Ala Arg Asn Ala Ala 85 90 95 Ile Ala Asn Leu Glu
Gly Leu Gly Asn Asn Phe Asn Ile Tyr Val Glu 100 105 110 Ala Phe Lys
Glu Trp Glu Glu Asp Pro Lys Asn Pro Ala Thr Arg Thr 115 120 125 Arg
Val Ile Asp Arg Phe Arg Ile Leu Asp Gly Leu Leu Glu Arg Asp 130 135
140 Ile Pro Ser Phe Arg Ile Ser Gly Phe Glu Val Pro Leu Leu Ser Val
145 150 155 160 Tyr Ala Gln Ala Ala Asn Leu His Leu Ala Ile Leu Arg
Asp Ser Val 165 170 175 Ile Phe Gly Glu Arg Trp Gly Leu Thr Thr Ile
Asn Val Asn Glu Asn 180 185 190 Tyr Asn Arg Leu Ile Arg His Ile Asp
Glu Tyr Ala Asp His Cys Ala 195 200 205 Asn Thr Tyr Asn Arg Gly Leu
Asn Asn Leu Pro Lys Ser Thr Tyr Gln 210 215 220 Asp Trp Ile Thr Tyr
Asn Arg Leu Arg Arg Asp Leu Thr Leu Thr Val 225 230 235 240 Leu Asp
Ile Ala Ala Phe Phe Pro Asn Tyr Asp Asn Arg Arg Tyr Pro 245 250 255
Ile Gln Pro Val Gly Gln Leu Thr Arg Glu Val Tyr Thr Asp Pro Leu 260
265 270 Ile Asn Phe Asn Pro Gln Leu Gln Ser Val Ala Gln Leu Pro Thr
Phe 275 280 285 Asn Val Met Glu Asn Ser Ala Ile Arg Asn Pro His Leu
Phe Asp Ile 290 295 300 Leu Asn Asn Leu Thr Ile Phe Thr Asp Trp Phe
Ser Val Gly Arg Asn 305 310 315 320 Phe Tyr Trp Gly Gly His Arg Val
Ile Ser Ser Leu Ile Gly Gly Gly 325 330 335 Asn Ile Thr Ser Pro Ile
Tyr Gly Arg Glu Ala Asn Gln Glu Pro Pro 340 345 350 Arg Ser Phe Thr
Phe Asn Gly Pro Val Phe Arg Thr Leu Ser Asn Pro 355 360 365 Thr Leu
Arg Leu Leu Gln Gln Pro Trp Pro Ala Pro Pro Phe Asn Leu 370 375 380
Arg Gly Val Glu Gly Val Glu Phe Ser Thr Pro Thr Asn Ser Phe Thr 385
390 395 400 Tyr Arg Gly Arg Gly Thr Val Asp Ser Leu Thr Glu Leu Pro
Pro Glu 405 410 415 Asp Asn Ser Val Pro Pro Arg Glu Gly Tyr Ser His
Arg Leu Cys His 420 425 430 Ala Thr Phe Val Gln Arg Ser Gly Thr Pro
Phe Leu Thr Thr Gly Val 435 440 445 Val Phe Ser Trp Thr His Arg Ser
Ala Thr Leu Thr Asn Thr Ile Asp 450 455 460 Pro Glu Arg Ile Asn Gln
Ile Pro Leu Val Lys Gly Phe Arg Val Trp 465 470 475 480 Gly Gly Thr
Ser Val Ile Thr Gly Pro Gly Phe Thr Gly Gly Asp Ile 485 490 495 Leu
Arg Arg Asn Thr Phe Gly Asp Phe Val Ser Leu Gln Val Asn Ile 500 505
510 Asn Ser Pro Ile Thr Gln Arg Tyr Arg Leu Arg Phe Arg Tyr Ala Ser
515 520 525 Ser Arg Asp Ala Arg Val Ile Val Leu Thr Gly Ala Ala Ser
Thr Gly 530 535 540 Val Gly Gly Gln Val Ser Val Asn Met Pro Leu Gln
Lys Thr Met Glu 545 550 555 560 Ile Gly Glu Asn Leu Thr Ser Arg Thr
Phe Arg Tyr Thr Asp Phe Ser 565 570 575 Asn Pro Phe Ser Phe Arg Ala
Asn Pro Asp Ile Ile Gly Ile Ser Glu 580 585 590 Gln Pro Leu Phe Gly
Ala Gly Ser Ile Ser Ser Gly Glu Leu Tyr Ile 595 600 605 Asp Lys Ile
Glu Ile Ile Leu Ala Asp Ala Thr Phe Glu 610 615 620
291866DNAArtificial SequenceSynthetic DNA sequence in accordance
with the invention encoding Bacillus thuringiensis Cry1Ca core
toxin using codons optimized for maize and with sequences
identified in Table 2 removed and Table 1 sequences are maintained
29atg gat aac aat ccg aac atc aat gag tgc atc ccg tac aac tgc ctg
48Met Asp Asn Asn Pro Asn Ile Asn Glu Cys Ile Pro Tyr Asn Cys Leu 1
5 10 15 agc aac ccg gaa gaa gtg ctg ttg gat gga gaa cgg ata tca act
ggc 96Ser Asn Pro Glu Glu Val Leu Leu Asp Gly Glu Arg Ile Ser Thr
Gly 20 25 30 aat tca tcc att gac att tct ctg tca ctt gtt cag ttt
ctg gtg tct 144Asn Ser Ser Ile Asp Ile Ser Leu Ser Leu Val Gln Phe
Leu Val Ser 35 40 45 aac ttc gtc ccc ggc gga gga ttc ctg gtt gga
tta ata gat ttc gta 192Asn Phe Val Pro Gly Gly Gly Phe Leu Val Gly
Leu Ile Asp Phe Val 50 55 60 tgg gga ata gtt ggc cct tct caa tgg
gac gca ttt cta gta caa att 240Trp Gly Ile Val Gly Pro Ser Gln Trp
Asp Ala Phe Leu Val Gln Ile 65 70 75 80 gaa caa tta att aat gaa aga
ata gct gaa ttt gct agg aac gct gct 288Glu Gln Leu Ile Asn Glu Arg
Ile Ala Glu Phe Ala Arg Asn Ala Ala 85 90 95 att gct aat tta gaa
gga tta gga aac aat ttc aac atc tat gtg gaa 336Ile Ala Asn Leu Glu
Gly Leu Gly Asn Asn Phe Asn Ile Tyr Val Glu 100 105 110 gca ttt aag
gaa tgg gaa gaa gat cct aag aat cca gca acg agg acc 384Ala Phe Lys
Glu Trp Glu Glu Asp Pro Lys Asn Pro Ala Thr Arg Thr 115 120 125 aga
gta att gat cgc ttt cgt ata ctt gat ggg cta ctt gaa agg gac 432Arg
Val Ile Asp Arg Phe Arg Ile Leu Asp Gly Leu Leu Glu Arg Asp 130 135
140 att cct tcg ttt cga att tct gga ttt gaa gta ccc ctt ctc tcc gtt
480Ile Pro Ser Phe Arg Ile Ser Gly Phe Glu Val Pro Leu Leu Ser Val
145 150 155 160 tat gct caa gcg gcc aat ctg cat cta gct atc tta aga
gat tct gtc 528Tyr Ala Gln Ala Ala Asn Leu His Leu Ala Ile Leu Arg
Asp Ser Val 165 170 175 atc ttt gga gaa aga tgg gga ttg aca acg ata
aat gtc aat gaa aac 576Ile Phe Gly Glu Arg Trp Gly Leu Thr Thr Ile
Asn Val Asn Glu Asn 180 185 190 tat aat aga cta att agg cat att gat
gaa tat gct gat cac tgt gca 624Tyr Asn Arg Leu Ile Arg His Ile Asp
Glu Tyr Ala Asp His Cys Ala 195 200 205 aat acg tat aat cgg gga tta
aat aat tta ccg aaa tct acg tat caa 672Asn Thr Tyr Asn Arg Gly Leu
Asn Asn Leu Pro Lys Ser Thr Tyr Gln 210 215 220 gat tgg ata aca tat
aat cga ttg cgg aga gac tta aca ttg act gta 720Asp Trp Ile Thr Tyr
Asn Arg Leu Arg Arg Asp Leu Thr Leu Thr Val 225 230 235 240 tta gat
atc gcc gct ttc ttt cca aac tat gac aat agg aga tat cca 768Leu Asp
Ile Ala Ala Phe Phe Pro Asn Tyr Asp Asn Arg Arg Tyr Pro 245 250 255
att cag cca gtt ggt caa cta aca agg gaa gtt tat acg gac cca tta
816Ile Gln Pro Val Gly Gln Leu Thr Arg Glu Val Tyr Thr Asp Pro Leu
260 265 270 att aat ttt aat cca cag tta cag tct gta gct caa tta cct
act ttt 864Ile Asn Phe Asn Pro Gln Leu Gln Ser Val Ala Gln Leu Pro
Thr Phe 275 280 285
aac gtt atg gag aac agc gca att aga aat cct cat ttg ttc gac ata
912Asn Val Met Glu Asn Ser Ala Ile Arg Asn Pro His Leu Phe Asp Ile
290 295 300 ttg aat aat ctt aca atc ttt acg gat tgg ttt agt gtt gga
cgc aac 960Leu Asn Asn Leu Thr Ile Phe Thr Asp Trp Phe Ser Val Gly
Arg Asn 305 310 315 320 ttc tat tgg gga gga cat cga gta ata tct agc
ctt ata gga ggt ggg 1008Phe Tyr Trp Gly Gly His Arg Val Ile Ser Ser
Leu Ile Gly Gly Gly 325 330 335 aac atc aca tcg cct atc tat gga aga
gag gcg aac caa gag cct cca 1056Asn Ile Thr Ser Pro Ile Tyr Gly Arg
Glu Ala Asn Gln Glu Pro Pro 340 345 350 aga tcc ttt act ttt aat gga
ccc gtg ttt agg act tta tca aat cct 1104Arg Ser Phe Thr Phe Asn Gly
Pro Val Phe Arg Thr Leu Ser Asn Pro 355 360 365 act tta cga tta tta
cag caa cct tgg cca gcg cca cca ttt aat tta 1152Thr Leu Arg Leu Leu
Gln Gln Pro Trp Pro Ala Pro Pro Phe Asn Leu 370 375 380 cgt ggt gtt
gaa gga gta gaa ttt tct aca cct aca aat agc ttt acg 1200Arg Gly Val
Glu Gly Val Glu Phe Ser Thr Pro Thr Asn Ser Phe Thr 385 390 395 400
tat cga gga aga ggg acg gtt gat tct tta act gaa ttg ccg cct gag
1248Tyr Arg Gly Arg Gly Thr Val Asp Ser Leu Thr Glu Leu Pro Pro Glu
405 410 415 gat aat agt gtg cca cct cgc gaa gga tat agt cat cgt tta
tgt cat 1296Asp Asn Ser Val Pro Pro Arg Glu Gly Tyr Ser His Arg Leu
Cys His 420 425 430 gca acc ttt gtt caa aga tcg gga aca cct ttc tta
aca act ggt gta 1344Ala Thr Phe Val Gln Arg Ser Gly Thr Pro Phe Leu
Thr Thr Gly Val 435 440 445 gta ttc tct tgg acg cat cgt agt gca act
ctt aca aat aca atc gac 1392Val Phe Ser Trp Thr His Arg Ser Ala Thr
Leu Thr Asn Thr Ile Asp 450 455 460 cca gag aga att aat caa ata cct
tta gtg aag gga ttt aga gtt tgg 1440Pro Glu Arg Ile Asn Gln Ile Pro
Leu Val Lys Gly Phe Arg Val Trp 465 470 475 480 ggg ggc acc tct gtc
att acc gga ccc gga ttt acc gga ggg gat atc 1488Gly Gly Thr Ser Val
Ile Thr Gly Pro Gly Phe Thr Gly Gly Asp Ile 485 490 495 ctt cga aga
aat acc ttt ggt gat ttc gta tct cta caa gtc aac att 1536Leu Arg Arg
Asn Thr Phe Gly Asp Phe Val Ser Leu Gln Val Asn Ile 500 505 510 aat
tca cca att acc caa aga tac cgt tta aga ttt cgt tac gct tcc 1584Asn
Ser Pro Ile Thr Gln Arg Tyr Arg Leu Arg Phe Arg Tyr Ala Ser 515 520
525 agt agg gat gca cga gtt ata gta tta acg gga gcg gca tcc acc gga
1632Ser Arg Asp Ala Arg Val Ile Val Leu Thr Gly Ala Ala Ser Thr Gly
530 535 540 gtg gga ggc caa gtt agt gta aat atg cct ctt cag aaa act
atg gaa 1680Val Gly Gly Gln Val Ser Val Asn Met Pro Leu Gln Lys Thr
Met Glu 545 550 555 560 ata ggg gag aac tta aca tcc aga aca ttt aga
tat acc gat ttt agt 1728Ile Gly Glu Asn Leu Thr Ser Arg Thr Phe Arg
Tyr Thr Asp Phe Ser 565 570 575 aat cct ttt tca ttt aga gct aat cca
gat ata att ggg ata agt gaa 1776Asn Pro Phe Ser Phe Arg Ala Asn Pro
Asp Ile Ile Gly Ile Ser Glu 580 585 590 caa cct cta ttt ggg gcg ggt
tct att agt agc ggt gaa ctt tac ata 1824Gln Pro Leu Phe Gly Ala Gly
Ser Ile Ser Ser Gly Glu Leu Tyr Ile 595 600 605 gat aaa att gaa att
att cta gca gat gca aca ttt gaa tga 1866Asp Lys Ile Glu Ile Ile Leu
Ala Asp Ala Thr Phe Glu 610 615 620 30621PRTArtificial
SequenceSynthetic Construct 30Met Asp Asn Asn Pro Asn Ile Asn Glu
Cys Ile Pro Tyr Asn Cys Leu 1 5 10 15 Ser Asn Pro Glu Glu Val Leu
Leu Asp Gly Glu Arg Ile Ser Thr Gly 20 25 30 Asn Ser Ser Ile Asp
Ile Ser Leu Ser Leu Val Gln Phe Leu Val Ser 35 40 45 Asn Phe Val
Pro Gly Gly Gly Phe Leu Val Gly Leu Ile Asp Phe Val 50 55 60 Trp
Gly Ile Val Gly Pro Ser Gln Trp Asp Ala Phe Leu Val Gln Ile 65 70
75 80 Glu Gln Leu Ile Asn Glu Arg Ile Ala Glu Phe Ala Arg Asn Ala
Ala 85 90 95 Ile Ala Asn Leu Glu Gly Leu Gly Asn Asn Phe Asn Ile
Tyr Val Glu 100 105 110 Ala Phe Lys Glu Trp Glu Glu Asp Pro Lys Asn
Pro Ala Thr Arg Thr 115 120 125 Arg Val Ile Asp Arg Phe Arg Ile Leu
Asp Gly Leu Leu Glu Arg Asp 130 135 140 Ile Pro Ser Phe Arg Ile Ser
Gly Phe Glu Val Pro Leu Leu Ser Val 145 150 155 160 Tyr Ala Gln Ala
Ala Asn Leu His Leu Ala Ile Leu Arg Asp Ser Val 165 170 175 Ile Phe
Gly Glu Arg Trp Gly Leu Thr Thr Ile Asn Val Asn Glu Asn 180 185 190
Tyr Asn Arg Leu Ile Arg His Ile Asp Glu Tyr Ala Asp His Cys Ala 195
200 205 Asn Thr Tyr Asn Arg Gly Leu Asn Asn Leu Pro Lys Ser Thr Tyr
Gln 210 215 220 Asp Trp Ile Thr Tyr Asn Arg Leu Arg Arg Asp Leu Thr
Leu Thr Val 225 230 235 240 Leu Asp Ile Ala Ala Phe Phe Pro Asn Tyr
Asp Asn Arg Arg Tyr Pro 245 250 255 Ile Gln Pro Val Gly Gln Leu Thr
Arg Glu Val Tyr Thr Asp Pro Leu 260 265 270 Ile Asn Phe Asn Pro Gln
Leu Gln Ser Val Ala Gln Leu Pro Thr Phe 275 280 285 Asn Val Met Glu
Asn Ser Ala Ile Arg Asn Pro His Leu Phe Asp Ile 290 295 300 Leu Asn
Asn Leu Thr Ile Phe Thr Asp Trp Phe Ser Val Gly Arg Asn 305 310 315
320 Phe Tyr Trp Gly Gly His Arg Val Ile Ser Ser Leu Ile Gly Gly Gly
325 330 335 Asn Ile Thr Ser Pro Ile Tyr Gly Arg Glu Ala Asn Gln Glu
Pro Pro 340 345 350 Arg Ser Phe Thr Phe Asn Gly Pro Val Phe Arg Thr
Leu Ser Asn Pro 355 360 365 Thr Leu Arg Leu Leu Gln Gln Pro Trp Pro
Ala Pro Pro Phe Asn Leu 370 375 380 Arg Gly Val Glu Gly Val Glu Phe
Ser Thr Pro Thr Asn Ser Phe Thr 385 390 395 400 Tyr Arg Gly Arg Gly
Thr Val Asp Ser Leu Thr Glu Leu Pro Pro Glu 405 410 415 Asp Asn Ser
Val Pro Pro Arg Glu Gly Tyr Ser His Arg Leu Cys His 420 425 430 Ala
Thr Phe Val Gln Arg Ser Gly Thr Pro Phe Leu Thr Thr Gly Val 435 440
445 Val Phe Ser Trp Thr His Arg Ser Ala Thr Leu Thr Asn Thr Ile Asp
450 455 460 Pro Glu Arg Ile Asn Gln Ile Pro Leu Val Lys Gly Phe Arg
Val Trp 465 470 475 480 Gly Gly Thr Ser Val Ile Thr Gly Pro Gly Phe
Thr Gly Gly Asp Ile 485 490 495 Leu Arg Arg Asn Thr Phe Gly Asp Phe
Val Ser Leu Gln Val Asn Ile 500 505 510 Asn Ser Pro Ile Thr Gln Arg
Tyr Arg Leu Arg Phe Arg Tyr Ala Ser 515 520 525 Ser Arg Asp Ala Arg
Val Ile Val Leu Thr Gly Ala Ala Ser Thr Gly 530 535 540 Val Gly Gly
Gln Val Ser Val Asn Met Pro Leu Gln Lys Thr Met Glu 545 550 555 560
Ile Gly Glu Asn Leu Thr Ser Arg Thr Phe Arg Tyr Thr Asp Phe Ser 565
570 575 Asn Pro Phe Ser Phe Arg Ala Asn Pro Asp Ile Ile Gly Ile Ser
Glu 580 585 590 Gln Pro Leu Phe Gly Ala Gly Ser Ile Ser Ser Gly Glu
Leu Tyr Ile 595 600 605 Asp Lys Ile Glu Ile Ile Leu Ala Asp Ala Thr
Phe Glu 610 615 620 311428DNABacillus
thuringiensisCDS(1)..(1428)Native DNA sequence encoding Bacillus
thuringiensis Cry6Aa toxin 31atg att att gat agt aaa acg act tta
cct aga cat tca ctt att cat 48Met Ile Ile Asp Ser Lys Thr Thr Leu
Pro Arg His Ser Leu Ile His 1 5 10 15 aca att aaa tta aat tct aat
aag aaa tat ggt cct ggt gat atg act 96Thr Ile Lys Leu Asn Ser Asn
Lys Lys Tyr Gly Pro Gly Asp Met Thr 20 25 30 aat gga aat caa ttt
att att tca aaa caa gaa tgg gct acg att gga 144Asn Gly Asn Gln Phe
Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly 35 40 45 gca tat att
cag act gga tta ggt tta cca gta aat gaa caa caa tta 192Ala Tyr Ile
Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu 50 55 60 aga
aca cat gtt aat tta agt cag gat ata tca ata cct agt gat ttt 240Arg
Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe 65 70
75 80 tct caa tta tat gat gtt tat tgt tct gat aaa act tca gca gaa
tgg 288Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu
Trp 85 90 95 tgg aat aaa aat tta tat cct tta att att aaa tct gct
aat gat att 336Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala
Asn Asp Ile 100 105 110 gct tca tat ggt ttt aaa gtt gct ggt gat cct
tct att aag aaa gat 384Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro
Ser Ile Lys Lys Asp 115 120 125 gga tat ttt aaa aaa ttg caa gat gaa
tta gat aat att gtt gat aat 432Gly Tyr Phe Lys Lys Leu Gln Asp Glu
Leu Asp Asn Ile Val Asp Asn 130 135 140 aat tcc gat gat gat gca ata
gct aaa gct att aaa gat ttt aaa gcg 480Asn Ser Asp Asp Asp Ala Ile
Ala Lys Ala Ile Lys Asp Phe Lys Ala 145 150 155 160 cga tgt ggt att
tta att aaa gaa gct aaa caa tat gaa gaa gct gca 528Arg Cys Gly Ile
Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala 165 170 175 aaa aat
att gta aca tct tta gat caa ttt tta cat ggt gat cag aaa 576Lys Asn
Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys 180 185 190
aaa tta gaa ggt gtt atc aat att caa aaa cgt tta aaa gaa gtt caa
624Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln
195 200 205 aca gct ctt aat caa gcc cat ggg gaa agt agt cca gct cat
aaa gag 672Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His
Lys Glu 210 215 220 tta tta gaa aaa gta aaa aat tta aaa aca aca tta
gaa agg act att 720Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu
Glu Arg Thr Ile 225 230 235 240 aaa gct gaa caa gat tta gag aaa aaa
gta gaa tat agt ttt cta tta 768Lys Ala Glu Gln Asp Leu Glu Lys Lys
Val Glu Tyr Ser Phe Leu Leu 245 250 255 gga cca ttg tta gga ttt gtt
gtt tat gaa att ctt gaa aat act gct 816Gly Pro Leu Leu Gly Phe Val
Val Tyr Glu Ile Leu Glu Asn Thr Ala 260 265 270 gtt cag cat ata aaa
aat caa att gat gag ata aag aaa caa tta gat 864Val Gln His Ile Lys
Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp 275 280 285 tct gct cag
cat gat ttg gat aga gat gtt aaa att ata gga atg tta 912Ser Ala Gln
His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu 290 295 300 aat
agt att aat aca gat att gat aat tta tat agt caa gga caa gaa 960Asn
Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu 305 310
315 320 gca att aaa gtt ttc caa aag tta caa ggt att tgg gct act att
gga 1008Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile
Gly 325 330 335 gct caa ata gaa aat ctt aga aca acg tcg tta caa gaa
gtt caa gat 1056Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu
Val Gln Asp 340 345 350 tct gat gat gct gat gag ata caa att gaa ctt
gag gac gct tct gat 1104Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu
Glu Asp Ala Ser Asp 355 360 365 gct tgg tta gtt gtg gct caa gaa gct
cgt gat ttt aca cta aat gct 1152Ala Trp Leu Val Val Ala Gln Glu Ala
Arg Asp Phe Thr Leu Asn Ala 370 375 380 tat tca act aat agt aga caa
aat tta ccg att aat gtt ata tca gat 1200Tyr Ser Thr Asn Ser Arg Gln
Asn Leu Pro Ile Asn Val Ile Ser Asp 385 390 395 400 tca tgt aat tgt
tca aca aca aat atg aca tca aat caa tac agt aat 1248Ser Cys Asn Cys
Ser Thr Thr Asn Met Thr Ser Asn Gln Tyr Ser Asn 405 410 415 cca aca
aca aat atg aca tca aat caa tat atg att tca cat gaa tat 1296Pro Thr
Thr Asn Met Thr Ser Asn Gln Tyr Met Ile Ser His Glu Tyr 420 425 430
aca agt tta cca aat aat ttt atg tta tca aga aat agt aat tta gaa
1344Thr Ser Leu Pro Asn Asn Phe Met Leu Ser Arg Asn Ser Asn Leu Glu
435 440 445 tat aaa tgt cct gaa aat aat ttt atg ata tat tgg tat aat
aat tcg 1392Tyr Lys Cys Pro Glu Asn Asn Phe Met Ile Tyr Trp Tyr Asn
Asn Ser 450 455 460 gat tgg tat aat aat tcg gat tgg tat aat aat tga
1428Asp Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn 465 470 475
32475PRTBacillus thuringiensis 32Met Ile Ile Asp Ser Lys Thr Thr
Leu Pro Arg His Ser Leu Ile His 1 5 10 15 Thr Ile Lys Leu Asn Ser
Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr 20 25 30 Asn Gly Asn Gln
Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly 35 40 45 Ala Tyr
Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu 50 55 60
Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe 65
70 75 80 Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala
Glu Trp 85 90 95 Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser
Ala Asn Asp Ile 100 105 110 Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp
Pro Ser Ile Lys Lys Asp 115 120 125 Gly Tyr Phe Lys Lys Leu Gln Asp
Glu Leu Asp Asn Ile Val Asp Asn 130 135 140 Asn Ser Asp Asp Asp Ala
Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala 145 150 155 160 Arg Cys Gly
Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala 165 170 175 Lys
Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys 180 185
190 Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln
195 200 205 Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His
Lys Glu 210 215 220 Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu
Glu Arg Thr Ile 225 230 235 240 Lys Ala Glu Gln Asp Leu Glu Lys Lys
Val Glu Tyr Ser Phe Leu Leu 245 250 255 Gly Pro Leu Leu Gly Phe Val
Val Tyr Glu Ile Leu Glu Asn
Thr Ala 260 265 270 Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys
Lys Gln Leu Asp 275 280 285 Ser Ala Gln His Asp Leu Asp Arg Asp Val
Lys Ile Ile Gly Met Leu 290 295 300 Asn Ser Ile Asn Thr Asp Ile Asp
Asn Leu Tyr Ser Gln Gly Gln Glu 305 310 315 320 Ala Ile Lys Val Phe
Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly 325 330 335 Ala Gln Ile
Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp 340 345 350 Ser
Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp 355 360
365 Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala
370 375 380 Tyr Ser Thr Asn Ser Arg Gln Asn Leu Pro Ile Asn Val Ile
Ser Asp 385 390 395 400 Ser Cys Asn Cys Ser Thr Thr Asn Met Thr Ser
Asn Gln Tyr Ser Asn 405 410 415 Pro Thr Thr Asn Met Thr Ser Asn Gln
Tyr Met Ile Ser His Glu Tyr 420 425 430 Thr Ser Leu Pro Asn Asn Phe
Met Leu Ser Arg Asn Ser Asn Leu Glu 435 440 445 Tyr Lys Cys Pro Glu
Asn Asn Phe Met Ile Tyr Trp Tyr Asn Asn Ser 450 455 460 Asp Trp Tyr
Asn Asn Ser Asp Trp Tyr Asn Asn 465 470 475 331428DNAArtificial
SequenceSynthetic DNA sequence encoding Bacillus thuringiensis
Cry6Aa toxin using codons optimized for maize and Table 1 sequences
are maintained 33atg atc atc gac tcc aag acg acc ctg cca cgg cac
tcc ctt atc cac 48Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His
Ser Leu Ile His 1 5 10 15 aca att aaa tta aat agc aat aag aag tac
ggt ccc ggt gat atg act 96Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr
Gly Pro Gly Asp Met Thr 20 25 30 aac gga aat caa ttc att att tca
aag caa gag tgg gct acc atc gga 144Asn Gly Asn Gln Phe Ile Ile Ser
Lys Gln Glu Trp Ala Thr Ile Gly 35 40 45 gcg tac atc cag act ggg
ctg ggc cta cca gta aat gaa caa caa tta 192Ala Tyr Ile Gln Thr Gly
Leu Gly Leu Pro Val Asn Glu Gln Gln Leu 50 55 60 agg acc cat gtc
aac ctc agc caa gat atc agc atc cct agc gac ttt 240Arg Thr His Val
Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe 65 70 75 80 tct cag
ctc tac gac gtc tat tgc agc gat aaa act tcc gca gaa tgg 288Ser Gln
Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp 85 90 95
tgg aat aaa aac ctg tac ccc ctc atc att aaa tct gcc aac gat att
336Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile
100 105 110 gcc agc tac ggc ttc aag gtc gcg ggt gat cct tct att aag
aag gac 384Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys
Lys Asp 115 120 125 ggc tac ttc aag aag ctg caa gat gag ctg gac aac
att gtt gac aat 432Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn
Ile Val Asp Asn 130 135 140 aat tcc gat gat gat gca ata gcg aaa gcc
att aaa gac ttc aag gcg 480Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala
Ile Lys Asp Phe Lys Ala 145 150 155 160 cga tgc ggc atc cta att aaa
gaa gca aag cag tat gaa gag gca gcg 528Arg Cys Gly Ile Leu Ile Lys
Glu Ala Lys Gln Tyr Glu Glu Ala Ala 165 170 175 aaa aat atc gta aca
tcc ctc gac caa ttt ctg cat ggc gat cag aag 576Lys Asn Ile Val Thr
Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys 180 185 190 aaa ttg gag
ggt gtg atc aac atc caa aaa cgt ctg aag gag gtg cag 624Lys Leu Glu
Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln 195 200 205 acg
gct ctt aat caa gcc cac ggg gaa agt tca cca gct cat aaa gag 672Thr
Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu 210 215
220 ctg tta gag aaa gtc aag aat ctc aag acc aca ctt gag agg acc att
720Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile
225 230 235 240 aaa gct gag caa gac ctg gag aag aaa gtg gag tac agt
ttc ctt ctc 768Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser
Phe Leu Leu 245 250 255 ggc ccc ttg ctg ggc ttc gtc gtt tat gaa atc
ctt gaa aat act gcc 816Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile
Leu Glu Asn Thr Ala 260 265 270 gtc cag cat ata aaa aac caa att gac
gag ata aag aag caa ctg gac 864Val Gln His Ile Lys Asn Gln Ile Asp
Glu Ile Lys Lys Gln Leu Asp 275 280 285 tct gcc cag cac gac ttg gac
aga gac gtt aag atc ata ggg atg ctg 912Ser Ala Gln His Asp Leu Asp
Arg Asp Val Lys Ile Ile Gly Met Leu 290 295 300 aac agt att aat aca
gac att gat aac ttg tat agc caa gga caa gag 960Asn Ser Ile Asn Thr
Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu 305 310 315 320 gca att
aaa gtg ttc caa aag ctc caa ggc atc tgg gca act atc gga 1008Ala Ile
Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly 325 330 335
gcg cag ata gag aac ctt agg aca acg tcg ctc caa gaa gtg caa gac
1056Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp
340 345 350 tct gac gac gcc gat gag atc caa att gaa ctt gag gac gcg
tct gat 1104Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala
Ser Asp 355 360 365 gct tgg tta gtg gtg gcc caa gaa gct cgc gac ttc
aca cta aat gcc 1152Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe
Thr Leu Asn Ala 370 375 380 tac tca act aac tcg cgt cag aat cta ccg
att aat gtt ata tcc gat 1200Tyr Ser Thr Asn Ser Arg Gln Asn Leu Pro
Ile Asn Val Ile Ser Asp 385 390 395 400 tcc tgc aac tgt tcc aca acg
aac atg acc tca aat caa tac agt aat 1248Ser Cys Asn Cys Ser Thr Thr
Asn Met Thr Ser Asn Gln Tyr Ser Asn 405 410 415 cca acc aca aat atg
acc tca aat caa tat atg atc tca cac gag tat 1296Pro Thr Thr Asn Met
Thr Ser Asn Gln Tyr Met Ile Ser His Glu Tyr 420 425 430 acc tcg ttg
ccg aat aat ttc atg ctc tca aga aat agc aat ctg gaa 1344Thr Ser Leu
Pro Asn Asn Phe Met Leu Ser Arg Asn Ser Asn Leu Glu 435 440 445 tat
aag tgt cct gaa aat aat ttc atg ata tac tgg tac aat aat tcg 1392Tyr
Lys Cys Pro Glu Asn Asn Phe Met Ile Tyr Trp Tyr Asn Asn Ser 450 455
460 gac tgg tac aat aat tcg gat tgg tac aat aat tga 1428Asp Trp Tyr
Asn Asn Ser Asp Trp Tyr Asn Asn 465 470 475 34475PRTArtificial
SequenceSynthetic Construct 34Met Ile Ile Asp Ser Lys Thr Thr Leu
Pro Arg His Ser Leu Ile His 1 5 10 15 Thr Ile Lys Leu Asn Ser Asn
Lys Lys Tyr Gly Pro Gly Asp Met Thr 20 25 30 Asn Gly Asn Gln Phe
Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly 35 40 45 Ala Tyr Ile
Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu 50 55 60 Arg
Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe 65 70
75 80 Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu
Trp 85 90 95 Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala
Asn Asp Ile 100 105 110 Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro
Ser Ile Lys Lys Asp 115 120 125 Gly Tyr Phe Lys Lys Leu Gln Asp Glu
Leu Asp Asn Ile Val Asp Asn 130 135 140 Asn Ser Asp Asp Asp Ala Ile
Ala Lys Ala Ile Lys Asp Phe Lys Ala 145 150 155 160 Arg Cys Gly Ile
Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala 165 170 175 Lys Asn
Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys 180 185 190
Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln 195
200 205 Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys
Glu 210 215 220 Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu
Arg Thr Ile 225 230 235 240 Lys Ala Glu Gln Asp Leu Glu Lys Lys Val
Glu Tyr Ser Phe Leu Leu 245 250 255 Gly Pro Leu Leu Gly Phe Val Val
Tyr Glu Ile Leu Glu Asn Thr Ala 260 265 270 Val Gln His Ile Lys Asn
Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp 275 280 285 Ser Ala Gln His
Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu 290 295 300 Asn Ser
Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu 305 310 315
320 Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly
325 330 335 Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val
Gln Asp 340 345 350 Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu
Asp Ala Ser Asp 355 360 365 Ala Trp Leu Val Val Ala Gln Glu Ala Arg
Asp Phe Thr Leu Asn Ala 370 375 380 Tyr Ser Thr Asn Ser Arg Gln Asn
Leu Pro Ile Asn Val Ile Ser Asp 385 390 395 400 Ser Cys Asn Cys Ser
Thr Thr Asn Met Thr Ser Asn Gln Tyr Ser Asn 405 410 415 Pro Thr Thr
Asn Met Thr Ser Asn Gln Tyr Met Ile Ser His Glu Tyr 420 425 430 Thr
Ser Leu Pro Asn Asn Phe Met Leu Ser Arg Asn Ser Asn Leu Glu 435 440
445 Tyr Lys Cys Pro Glu Asn Asn Phe Met Ile Tyr Trp Tyr Asn Asn Ser
450 455 460 Asp Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn 465 470 475
351428DNAArtificial SequenceSynthetic DNA sequence in accordance
with the invention encoding Bacillus thuringiensis Cry6Aa toxin
using codons optimized for maize and with sequences identified in
Table 2 removed and Table 1 sequences are maintained 35atg atc atc
gac tcc aag acg acc ctg cca cgg cac tcc ctt atc cac 48Met Ile Ile
Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His 1 5 10 15 aca
att aaa tta aat agc aat aag aag tac ggt ccc ggt gat atg act 96Thr
Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr 20 25
30 aac gga aat caa ttc att atc tca aag caa gag tgg gct acc atc gga
144Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly
35 40 45 gcg tac atc cag act ggg ctg ggc cta cca gta aat gaa caa
caa tta 192Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln
Gln Leu 50 55 60 agg acc cat gtc aac ctc agc caa gat atc agc atc
cct agc gac ttt 240Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile
Pro Ser Asp Phe 65 70 75 80 tct cag ctc tac gac gtc tat tgc agc gat
aaa act tcc gca gaa tgg 288Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp
Lys Thr Ser Ala Glu Trp 85 90 95 tgg aat aaa aac ctg tac ccc ctc
atc att aaa tct gcc aac gat att 336Trp Asn Lys Asn Leu Tyr Pro Leu
Ile Ile Lys Ser Ala Asn Asp Ile 100 105 110 gcc agc tac ggc ttc aag
gtc gcg ggt gat cct tct att aag aag gac 384Ala Ser Tyr Gly Phe Lys
Val Ala Gly Asp Pro Ser Ile Lys Lys Asp 115 120 125 ggc tac ttc aag
aag ctg caa gat gag ctg gac aac att gtt gac aat 432Gly Tyr Phe Lys
Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn 130 135 140 aat tcc
gat gat gat gca ata gcg aaa gcc att aaa gac ttc aag gcg 480Asn Ser
Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala 145 150 155
160 cga tgc ggc atc cta att aaa gaa gca aag cag tat gaa gag gca gcg
528Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala
165 170 175 aaa aat atc gta aca tcc ctc gac caa ttt ctg cat ggc gat
cag aag 576Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp
Gln Lys 180 185 190 aaa ttg gag ggt gtg atc aac atc caa aaa cgt ctg
aag gag gtg cag 624Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu
Lys Glu Val Gln 195 200 205 acg gct ctt aat caa gcc cac ggg gaa agt
tca cca gct cat aaa gag 672Thr Ala Leu Asn Gln Ala His Gly Glu Ser
Ser Pro Ala His Lys Glu 210 215 220 ctg tta gag aaa gtc aag aat ctc
aag acc aca ctt gag agg acc att 720Leu Leu Glu Lys Val Lys Asn Leu
Lys Thr Thr Leu Glu Arg Thr Ile 225 230 235 240 aaa gct gag caa gac
ctg gag aag aaa gtg gag tac agt ttc ctt ctc 768Lys Ala Glu Gln Asp
Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu 245 250 255 ggc ccc ttg
ctg ggc ttc gtc gtt tat gaa atc ctt gaa aat act gcc 816Gly Pro Leu
Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala 260 265 270 gtc
cag cat ata aaa aac caa att gac gag ata aag aag caa ctg gac 864Val
Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp 275 280
285 tct gcc cag cac gac ttg gac aga gac gtt aag atc ata ggg atg ctg
912Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu
290 295 300 aac agt att aat aca gac att gat aac ttg tat agc caa gga
caa gag 960Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly
Gln Glu 305 310 315 320 gca att aaa gtg ttc caa aag ctc caa ggc atc
tgg gca act atc gga 1008Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile
Trp Ala Thr Ile Gly 325 330 335 gcg cag ata gag aac ctt agg aca acg
tcg ctc caa gaa gtg caa gac 1056Ala Gln Ile Glu Asn Leu Arg Thr Thr
Ser Leu Gln Glu Val Gln Asp 340 345 350 tct gac gac gcc gat gag atc
caa att gaa ctt gag gac gcg tct gat 1104Ser Asp Asp Ala Asp Glu Ile
Gln Ile Glu Leu Glu Asp Ala Ser Asp 355 360 365 gct tgg tta gtg gtg
gcc caa gaa gct cgc gac ttc aca cta aat gcc 1152Ala Trp Leu Val Val
Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala 370 375 380 tac tca act
aac tcg cgt cag aat cta ccg att aat gtt atc tcc gat 1200Tyr Ser Thr
Asn Ser Arg Gln Asn Leu Pro Ile Asn Val Ile Ser Asp 385 390 395 400
tcc tgc aac tgt tcc aca acg aac atg acc tca aat caa tac agt aat
1248Ser Cys Asn Cys Ser Thr Thr Asn Met Thr Ser Asn Gln Tyr Ser Asn
405 410 415 cca acc aca aat atg acc tca aat caa tac atg atc tca cac
gag tat 1296Pro Thr Thr Asn Met Thr Ser Asn Gln Tyr Met Ile Ser His
Glu Tyr 420 425 430
acc tcg ttg ccg aat aat ttc atg ctc tca aga aat agc aat ctg gaa
1344Thr Ser Leu Pro Asn Asn Phe Met Leu Ser Arg Asn Ser Asn Leu Glu
435 440 445 tat aag tgt cct gaa aat aat ttc atg ata tac tgg tac aat
aat tcg 1392Tyr Lys Cys Pro Glu Asn Asn Phe Met Ile Tyr Trp Tyr Asn
Asn Ser 450 455 460 gac tgg tac aat aat tcg gat tgg tac aat aat tga
1428Asp Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn 465 470 475
36475PRTArtificial SequenceSynthetic Construct 36Met Ile Ile Asp
Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His 1 5 10 15 Thr Ile
Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr 20 25 30
Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly 35
40 45 Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln
Leu 50 55 60 Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro
Ser Asp Phe 65 70 75 80 Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys
Thr Ser Ala Glu Trp 85 90 95 Trp Asn Lys Asn Leu Tyr Pro Leu Ile
Ile Lys Ser Ala Asn Asp Ile 100 105 110 Ala Ser Tyr Gly Phe Lys Val
Ala Gly Asp Pro Ser Ile Lys Lys Asp 115 120 125 Gly Tyr Phe Lys Lys
Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn 130 135 140 Asn Ser Asp
Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala 145 150 155 160
Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala 165
170 175 Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln
Lys 180 185 190 Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys
Glu Val Gln 195 200 205 Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser
Pro Ala His Lys Glu 210 215 220 Leu Leu Glu Lys Val Lys Asn Leu Lys
Thr Thr Leu Glu Arg Thr Ile 225 230 235 240 Lys Ala Glu Gln Asp Leu
Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu 245 250 255 Gly Pro Leu Leu
Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala 260 265 270 Val Gln
His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp 275 280 285
Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu 290
295 300 Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln
Glu 305 310 315 320 Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp
Ala Thr Ile Gly 325 330 335 Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser
Leu Gln Glu Val Gln Asp 340 345 350 Ser Asp Asp Ala Asp Glu Ile Gln
Ile Glu Leu Glu Asp Ala Ser Asp 355 360 365 Ala Trp Leu Val Val Ala
Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala 370 375 380 Tyr Ser Thr Asn
Ser Arg Gln Asn Leu Pro Ile Asn Val Ile Ser Asp 385 390 395 400 Ser
Cys Asn Cys Ser Thr Thr Asn Met Thr Ser Asn Gln Tyr Ser Asn 405 410
415 Pro Thr Thr Asn Met Thr Ser Asn Gln Tyr Met Ile Ser His Glu Tyr
420 425 430 Thr Ser Leu Pro Asn Asn Phe Met Leu Ser Arg Asn Ser Asn
Leu Glu 435 440 445 Tyr Lys Cys Pro Glu Asn Asn Phe Met Ile Tyr Trp
Tyr Asn Asn Ser 450 455 460 Asp Trp Tyr Asn Asn Ser Asp Trp Tyr Asn
Asn 465 470 475 37888DNASphingobiurn
herbicidovoransCDS(1)..(888)Native DNA sequence encoding
Sphingobiurn herbicidovorans AAD1 protein 37atg cat gct gca ctg tcc
ccc ctc tcc cag cgc ttt gag cgc atc gcg 48Met His Ala Ala Leu Ser
Pro Leu Ser Gln Arg Phe Glu Arg Ile Ala 1 5 10 15 gtc cag ccg ctg
acc ggc gtc ctg ggc gcc gag atc acc ggc gtc gac 96Val Gln Pro Leu
Thr Gly Val Leu Gly Ala Glu Ile Thr Gly Val Asp 20 25 30 ctg cgc
gag ccg ctc gac gac agc acc tgg aac gaa atc ctc gac gcg 144Leu Arg
Glu Pro Leu Asp Asp Ser Thr Trp Asn Glu Ile Leu Asp Ala 35 40 45
ttc cac act tac cag gtc atc tat ttt ccc ggc cag gcg atc acc aac
192Phe His Thr Tyr Gln Val Ile Tyr Phe Pro Gly Gln Ala Ile Thr Asn
50 55 60 gaa cag cac atc gcc ttc agc cgg cgc ttc ggc ccc gtc gat
ccc gtg 240Glu Gln His Ile Ala Phe Ser Arg Arg Phe Gly Pro Val Asp
Pro Val 65 70 75 80 ccc ctg ctc aag agc atc gaa ggg tat cca gag gtg
cag atg atc cgc 288Pro Leu Leu Lys Ser Ile Glu Gly Tyr Pro Glu Val
Gln Met Ile Arg 85 90 95 cgc gaa gcc aac gaa agc ggg cgt gtg atc
ggt gat gac tgg cac acc 336Arg Glu Ala Asn Glu Ser Gly Arg Val Ile
Gly Asp Asp Trp His Thr 100 105 110 gac agc acc ttc ctg gac gca ccg
ccg gcc gcc gtg gtg atg cgc gcg 384Asp Ser Thr Phe Leu Asp Ala Pro
Pro Ala Ala Val Val Met Arg Ala 115 120 125 atc gac gtg ccc gag cat
ggc ggc gac acc ggt ttt ctg agc atg tac 432Ile Asp Val Pro Glu His
Gly Gly Asp Thr Gly Phe Leu Ser Met Tyr 130 135 140 acc gcg tgg gag
acg ctg tcg ccc acc atg cag gcc acc atc gaa ggg 480Thr Ala Trp Glu
Thr Leu Ser Pro Thr Met Gln Ala Thr Ile Glu Gly 145 150 155 160 ttg
aac gta gtg cac agc gcc acg cgt gtg ttc ggc tcg ctc tac cag 528Leu
Asn Val Val His Ser Ala Thr Arg Val Phe Gly Ser Leu Tyr Gln 165 170
175 gcc cag aac cgg cgc ttc agc aac acc agc gtc aag gtg atg gac gtc
576Ala Gln Asn Arg Arg Phe Ser Asn Thr Ser Val Lys Val Met Asp Val
180 185 190 gac gcg ggc gac cgt gaa acc gtg cac ccc ctg gtg gtg acc
cat ccg 624Asp Ala Gly Asp Arg Glu Thr Val His Pro Leu Val Val Thr
His Pro 195 200 205 ggc agc ggc cgc aag ggc ctg tac gtg aac cag gtc
tat tgc cag cgc 672Gly Ser Gly Arg Lys Gly Leu Tyr Val Asn Gln Val
Tyr Cys Gln Arg 210 215 220 atc gag ggc atg acc gat gcc gaa agc aaa
ccg ctg ctg cag ttc ctg 720Ile Glu Gly Met Thr Asp Ala Glu Ser Lys
Pro Leu Leu Gln Phe Leu 225 230 235 240 tac gag cat gcg aca cgg ttc
gat ttc acc tgc cgc gtg cgc tgg aag 768Tyr Glu His Ala Thr Arg Phe
Asp Phe Thr Cys Arg Val Arg Trp Lys 245 250 255 aag gac cag gtc ctg
gtc tgg gac aac ctg tgc acg atg cac cgg gcc 816Lys Asp Gln Val Leu
Val Trp Asp Asn Leu Cys Thr Met His Arg Ala 260 265 270 gta ccc gac
tac gcg ggc aag ttc cgc tac ctg acg cgc acc acg gtc 864Val Pro Asp
Tyr Ala Gly Lys Phe Arg Tyr Leu Thr Arg Thr Thr Val 275 280 285 ggt
ggc gtg cgc ccg gcg cgc tag 888Gly Gly Val Arg Pro Ala Arg 290 295
38295PRTSphingobiurn herbicidovorans 38Met His Ala Ala Leu Ser Pro
Leu Ser Gln Arg Phe Glu Arg Ile Ala 1 5 10 15 Val Gln Pro Leu Thr
Gly Val Leu Gly Ala Glu Ile Thr Gly Val Asp 20 25 30 Leu Arg Glu
Pro Leu Asp Asp Ser Thr Trp Asn Glu Ile Leu Asp Ala 35 40 45 Phe
His Thr Tyr Gln Val Ile Tyr Phe Pro Gly Gln Ala Ile Thr Asn 50 55
60 Glu Gln His Ile Ala Phe Ser Arg Arg Phe Gly Pro Val Asp Pro Val
65 70 75 80 Pro Leu Leu Lys Ser Ile Glu Gly Tyr Pro Glu Val Gln Met
Ile Arg 85 90 95 Arg Glu Ala Asn Glu Ser Gly Arg Val Ile Gly Asp
Asp Trp His Thr 100 105 110 Asp Ser Thr Phe Leu Asp Ala Pro Pro Ala
Ala Val Val Met Arg Ala 115 120 125 Ile Asp Val Pro Glu His Gly Gly
Asp Thr Gly Phe Leu Ser Met Tyr 130 135 140 Thr Ala Trp Glu Thr Leu
Ser Pro Thr Met Gln Ala Thr Ile Glu Gly 145 150 155 160 Leu Asn Val
Val His Ser Ala Thr Arg Val Phe Gly Ser Leu Tyr Gln 165 170 175 Ala
Gln Asn Arg Arg Phe Ser Asn Thr Ser Val Lys Val Met Asp Val 180 185
190 Asp Ala Gly Asp Arg Glu Thr Val His Pro Leu Val Val Thr His Pro
195 200 205 Gly Ser Gly Arg Lys Gly Leu Tyr Val Asn Gln Val Tyr Cys
Gln Arg 210 215 220 Ile Glu Gly Met Thr Asp Ala Glu Ser Lys Pro Leu
Leu Gln Phe Leu 225 230 235 240 Tyr Glu His Ala Thr Arg Phe Asp Phe
Thr Cys Arg Val Arg Trp Lys 245 250 255 Lys Asp Gln Val Leu Val Trp
Asp Asn Leu Cys Thr Met His Arg Ala 260 265 270 Val Pro Asp Tyr Ala
Gly Lys Phe Arg Tyr Leu Thr Arg Thr Thr Val 275 280 285 Gly Gly Val
Arg Pro Ala Arg 290 295 39888DNAArtificial SequenceSynthetic DNA
sequence encoding the AAD1 protein using codons optimized for maize
and Table 1 and Table 2 sequences are maintained 39atg cac gct gca
ctg tca cca ctc tca cag cgc ttt gag aga att gcg 48Met His Ala Ala
Leu Ser Pro Leu Ser Gln Arg Phe Glu Arg Ile Ala 1 5 10 15 gtc cag
ccg ctg act ggc gtc ttg ggc gct gag atc acc ggc gtc gat 96Val Gln
Pro Leu Thr Gly Val Leu Gly Ala Glu Ile Thr Gly Val Asp 20 25 30
ctg agg gag cct ctc gac gat tca acg tgg aac gaa att ctc gac gcg
144Leu Arg Glu Pro Leu Asp Asp Ser Thr Trp Asn Glu Ile Leu Asp Ala
35 40 45 ttc cat act tac caa gtc atc tat ttt ccc ggg caa gct att
acc aac 192Phe His Thr Tyr Gln Val Ile Tyr Phe Pro Gly Gln Ala Ile
Thr Asn 50 55 60 gaa caa cac atc gct ttc tct cgg cga ttc ggc ccc
gtc gat cca gtg 240Glu Gln His Ile Ala Phe Ser Arg Arg Phe Gly Pro
Val Asp Pro Val 65 70 75 80 ccc tta ctc aag tct atc gaa ggc tac cca
gag gtg cag atg ata aga 288Pro Leu Leu Lys Ser Ile Glu Gly Tyr Pro
Glu Val Gln Met Ile Arg 85 90 95 agg gag gcc aac gaa agc ggg cgt
gtg ata ggt gat gac tgg cac act 336Arg Glu Ala Asn Glu Ser Gly Arg
Val Ile Gly Asp Asp Trp His Thr 100 105 110 gac agc aca ttc ctg gat
gca ccg ccg gcc gct gtg gtg atg agg gca 384Asp Ser Thr Phe Leu Asp
Ala Pro Pro Ala Ala Val Val Met Arg Ala 115 120 125 atc gac gtg ccc
gag cac gga ggt gac act ggt ttc ttg agt atg tac 432Ile Asp Val Pro
Glu His Gly Gly Asp Thr Gly Phe Leu Ser Met Tyr 130 135 140 act gct
tgg gag acg ctt tcg cct act atg caa gcc aca atc gag ggg 480Thr Ala
Trp Glu Thr Leu Ser Pro Thr Met Gln Ala Thr Ile Glu Gly 145 150 155
160 ttg aat gta gtt cac agc gcc acg cgt gtg ttc gga tct ctc tat caa
528Leu Asn Val Val His Ser Ala Thr Arg Val Phe Gly Ser Leu Tyr Gln
165 170 175 gcc caa aac cgg cgc ttt tca aat acc tcc gtc aag gtg atg
gac gtt 576Ala Gln Asn Arg Arg Phe Ser Asn Thr Ser Val Lys Val Met
Asp Val 180 185 190 gac gcg ggc gac cgt gaa acc gtg cac cct ctt gtt
gta acc cat ccg 624Asp Ala Gly Asp Arg Glu Thr Val His Pro Leu Val
Val Thr His Pro 195 200 205 ggc agt ggt cgc aag ggc cta tac gtt aac
caa gtc tat tgc cag cgc 672Gly Ser Gly Arg Lys Gly Leu Tyr Val Asn
Gln Val Tyr Cys Gln Arg 210 215 220 atc gag gga atg aca gac gca gag
agt aag ccg ctc ctg caa ttc ctg 720Ile Glu Gly Met Thr Asp Ala Glu
Ser Lys Pro Leu Leu Gln Phe Leu 225 230 235 240 tac gag cac gcg aca
cgg ttc gat ttc acc tgc cgc gtg cgc tgg aaa 768Tyr Glu His Ala Thr
Arg Phe Asp Phe Thr Cys Arg Val Arg Trp Lys 245 250 255 aag gat caa
gtc ctt gta tgg gac aac ctt tgt acg atg cac cgg gcc 816Lys Asp Gln
Val Leu Val Trp Asp Asn Leu Cys Thr Met His Arg Ala 260 265 270 gtt
cct gac tac gcg ggc aag ttc aga tac ctg acg agg acc acg gtc 864Val
Pro Asp Tyr Ala Gly Lys Phe Arg Tyr Leu Thr Arg Thr Thr Val 275 280
285 ggt gga gtt agg cca gcg aga tga 888Gly Gly Val Arg Pro Ala Arg
290 295 40295PRTArtificial SequenceSynthetic Construct 40Met His
Ala Ala Leu Ser Pro Leu Ser Gln Arg Phe Glu Arg Ile Ala 1 5 10 15
Val Gln Pro Leu Thr Gly Val Leu Gly Ala Glu Ile Thr Gly Val Asp 20
25 30 Leu Arg Glu Pro Leu Asp Asp Ser Thr Trp Asn Glu Ile Leu Asp
Ala 35 40 45 Phe His Thr Tyr Gln Val Ile Tyr Phe Pro Gly Gln Ala
Ile Thr Asn 50 55 60 Glu Gln His Ile Ala Phe Ser Arg Arg Phe Gly
Pro Val Asp Pro Val 65 70 75 80 Pro Leu Leu Lys Ser Ile Glu Gly Tyr
Pro Glu Val Gln Met Ile Arg 85 90 95 Arg Glu Ala Asn Glu Ser Gly
Arg Val Ile Gly Asp Asp Trp His Thr 100 105 110 Asp Ser Thr Phe Leu
Asp Ala Pro Pro Ala Ala Val Val Met Arg Ala 115 120 125 Ile Asp Val
Pro Glu His Gly Gly Asp Thr Gly Phe Leu Ser Met Tyr 130 135 140 Thr
Ala Trp Glu Thr Leu Ser Pro Thr Met Gln Ala Thr Ile Glu Gly 145 150
155 160 Leu Asn Val Val His Ser Ala Thr Arg Val Phe Gly Ser Leu Tyr
Gln 165 170 175 Ala Gln Asn Arg Arg Phe Ser Asn Thr Ser Val Lys Val
Met Asp Val 180 185 190 Asp Ala Gly Asp Arg Glu Thr Val His Pro Leu
Val Val Thr His Pro 195 200 205 Gly Ser Gly Arg Lys Gly Leu Tyr Val
Asn Gln Val Tyr Cys Gln Arg 210 215 220 Ile Glu Gly Met Thr Asp Ala
Glu Ser Lys Pro Leu Leu Gln Phe Leu 225 230 235 240 Tyr Glu His Ala
Thr Arg Phe Asp Phe Thr Cys Arg Val Arg Trp Lys 245 250 255 Lys Asp
Gln Val Leu Val Trp Asp Asn Leu Cys Thr Met His Arg Ala 260 265 270
Val Pro Asp Tyr Ala Gly Lys Phe Arg Tyr Leu Thr Arg Thr Thr Val 275
280 285 Gly Gly Val Arg Pro Ala Arg 290 295 41888DNAArtificial
SequenceSynthetic DNA sequence encoding the AAD1 protein using
codons optimized for maize and with sequences identified in Table 2
removed and Table 1 sequences are maintained 41atg cac gct gca ctg
tca cca ctc tca cag cgc ttt gag aga att gcg 48Met His Ala Ala Leu
Ser Pro Leu Ser Gln Arg Phe Glu Arg Ile Ala 1 5 10 15 gtc cag
ccg ctg act ggc gtc ttg ggc gct gag atc acc ggc gtc gat 96Val Gln
Pro Leu Thr Gly Val Leu Gly Ala Glu Ile Thr Gly Val Asp 20 25 30
ctg agg gag cct ctc gac gat tca acg tgg aac gaa att ctc gac gcg
144Leu Arg Glu Pro Leu Asp Asp Ser Thr Trp Asn Glu Ile Leu Asp Ala
35 40 45 ttc cat act tac caa gtc atc tac ttt ccc ggg caa gct att
acc aac 192Phe His Thr Tyr Gln Val Ile Tyr Phe Pro Gly Gln Ala Ile
Thr Asn 50 55 60 gaa caa cac atc gct ttc tct cgg cga ttc ggc ccc
gtc gat cca gtg 240Glu Gln His Ile Ala Phe Ser Arg Arg Phe Gly Pro
Val Asp Pro Val 65 70 75 80 ccc tta ctc aag tct atc gaa ggc tac cca
gag gtg cag atg ata aga 288Pro Leu Leu Lys Ser Ile Glu Gly Tyr Pro
Glu Val Gln Met Ile Arg 85 90 95 agg gag gcc aac gaa agc ggg cgt
gtg ata ggt gat gac tgg cac act 336Arg Glu Ala Asn Glu Ser Gly Arg
Val Ile Gly Asp Asp Trp His Thr 100 105 110 gac agc aca ttc ctg gat
gca ccg ccg gcc gct gtg gtg atg agg gca 384Asp Ser Thr Phe Leu Asp
Ala Pro Pro Ala Ala Val Val Met Arg Ala 115 120 125 atc gac gtg ccc
gag cac gga ggt gac act ggt ttc ttg agt atg tac 432Ile Asp Val Pro
Glu His Gly Gly Asp Thr Gly Phe Leu Ser Met Tyr 130 135 140 act gct
tgg gag acg ctt tcg cct act atg caa gcc aca atc gag ggg 480Thr Ala
Trp Glu Thr Leu Ser Pro Thr Met Gln Ala Thr Ile Glu Gly 145 150 155
160 ttg aat gta gtt cac agc gcc acg cgt gtg ttc gga tct ctc tat caa
528Leu Asn Val Val His Ser Ala Thr Arg Val Phe Gly Ser Leu Tyr Gln
165 170 175 gcc caa aac cgg cgc ttt tca aat acc tcc gtc aag gtg atg
gac gtt 576Ala Gln Asn Arg Arg Phe Ser Asn Thr Ser Val Lys Val Met
Asp Val 180 185 190 gac gcg ggc gac cgt gaa acc gtg cac cct ctt gtt
gta acc cat ccg 624Asp Ala Gly Asp Arg Glu Thr Val His Pro Leu Val
Val Thr His Pro 195 200 205 ggc agt ggt cgc aag ggc cta tac gtt aac
caa gtc tat tgc cag cgc 672Gly Ser Gly Arg Lys Gly Leu Tyr Val Asn
Gln Val Tyr Cys Gln Arg 210 215 220 atc gag gga atg aca gac gca gag
agt aag ccg ctc ctg caa ttc ctg 720Ile Glu Gly Met Thr Asp Ala Glu
Ser Lys Pro Leu Leu Gln Phe Leu 225 230 235 240 tac gag cac gcg aca
cgg ttc gat ttc acc tgc cgc gtg cgc tgg aaa 768Tyr Glu His Ala Thr
Arg Phe Asp Phe Thr Cys Arg Val Arg Trp Lys 245 250 255 aag gat caa
gtc ctt gta tgg gac aac ctt tgt acg atg cac cgg gcc 816Lys Asp Gln
Val Leu Val Trp Asp Asn Leu Cys Thr Met His Arg Ala 260 265 270 gtt
cct gac tac gcg ggc aag ttc aga tac ctg acg agg acc acg gtc 864Val
Pro Asp Tyr Ala Gly Lys Phe Arg Tyr Leu Thr Arg Thr Thr Val 275 280
285 ggt gga gtt agg cca gcg aga tga 888Gly Gly Val Arg Pro Ala Arg
290 295 42295PRTArtificial SequenceSynthetic Construct 42Met His
Ala Ala Leu Ser Pro Leu Ser Gln Arg Phe Glu Arg Ile Ala 1 5 10 15
Val Gln Pro Leu Thr Gly Val Leu Gly Ala Glu Ile Thr Gly Val Asp 20
25 30 Leu Arg Glu Pro Leu Asp Asp Ser Thr Trp Asn Glu Ile Leu Asp
Ala 35 40 45 Phe His Thr Tyr Gln Val Ile Tyr Phe Pro Gly Gln Ala
Ile Thr Asn 50 55 60 Glu Gln His Ile Ala Phe Ser Arg Arg Phe Gly
Pro Val Asp Pro Val 65 70 75 80 Pro Leu Leu Lys Ser Ile Glu Gly Tyr
Pro Glu Val Gln Met Ile Arg 85 90 95 Arg Glu Ala Asn Glu Ser Gly
Arg Val Ile Gly Asp Asp Trp His Thr 100 105 110 Asp Ser Thr Phe Leu
Asp Ala Pro Pro Ala Ala Val Val Met Arg Ala 115 120 125 Ile Asp Val
Pro Glu His Gly Gly Asp Thr Gly Phe Leu Ser Met Tyr 130 135 140 Thr
Ala Trp Glu Thr Leu Ser Pro Thr Met Gln Ala Thr Ile Glu Gly 145 150
155 160 Leu Asn Val Val His Ser Ala Thr Arg Val Phe Gly Ser Leu Tyr
Gln 165 170 175 Ala Gln Asn Arg Arg Phe Ser Asn Thr Ser Val Lys Val
Met Asp Val 180 185 190 Asp Ala Gly Asp Arg Glu Thr Val His Pro Leu
Val Val Thr His Pro 195 200 205 Gly Ser Gly Arg Lys Gly Leu Tyr Val
Asn Gln Val Tyr Cys Gln Arg 210 215 220 Ile Glu Gly Met Thr Asp Ala
Glu Ser Lys Pro Leu Leu Gln Phe Leu 225 230 235 240 Tyr Glu His Ala
Thr Arg Phe Asp Phe Thr Cys Arg Val Arg Trp Lys 245 250 255 Lys Asp
Gln Val Leu Val Trp Asp Asn Leu Cys Thr Met His Arg Ala 260 265 270
Val Pro Asp Tyr Ala Gly Lys Phe Arg Tyr Leu Thr Arg Thr Thr Val 275
280 285 Gly Gly Val Arg Pro Ala Arg 290 295 431368DNAAspergillus
nidulansCDS(1)..(1368)Native DNA sequence encoding Aspergillus
nidulans delta-9 fatty acid desaturase protein 43atg tct gca cca
acg gcg gac atc agg gct cgc gcc ccg gag gcc aaa 48Met Ser Ala Pro
Thr Ala Asp Ile Arg Ala Arg Ala Pro Glu Ala Lys 1 5 10 15 aag gtt
cac atc gct gac act gct atc aac cgc cat aac tgg tac aag 96Lys Val
His Ile Ala Asp Thr Ala Ile Asn Arg His Asn Trp Tyr Lys 20 25 30
cat gtg aac tgg ctg aac gtt ttc ctg atc atc ggt atc ccg ctt tat
144His Val Asn Trp Leu Asn Val Phe Leu Ile Ile Gly Ile Pro Leu Tyr
35 40 45 ggg tgc att cag gcg ttc tgg gtg cca ctg cag ctg aag act
gcc atc 192Gly Cys Ile Gln Ala Phe Trp Val Pro Leu Gln Leu Lys Thr
Ala Ile 50 55 60 tgg gcc gtc atc tac tac ttt ttc acc ggt ctc ggt
atc aca gca ggt 240Trp Ala Val Ile Tyr Tyr Phe Phe Thr Gly Leu Gly
Ile Thr Ala Gly 65 70 75 80 tac cat cgt cta tgg gct cac tgc tcg tac
tcc gcc acc ctt cct ttg 288Tyr His Arg Leu Trp Ala His Cys Ser Tyr
Ser Ala Thr Leu Pro Leu 85 90 95 cgt atc tgg ctc gct gcc gtt ggt
ggt ggt gcc gtc gaa ggt tct atc 336Arg Ile Trp Leu Ala Ala Val Gly
Gly Gly Ala Val Glu Gly Ser Ile 100 105 110 cgc tgg tgg gct cgt gac
cac cgc gct cac cac cgc tac acc gat acc 384Arg Trp Trp Ala Arg Asp
His Arg Ala His His Arg Tyr Thr Asp Thr 115 120 125 gac aaa gac ccg
tac tcc gtt cgc aag ggt ctg ctc tac tct cac ctt 432Asp Lys Asp Pro
Tyr Ser Val Arg Lys Gly Leu Leu Tyr Ser His Leu 130 135 140 ggc tgg
atg gtg atg aag cag aac cct aag cgt att ggc cgt acc gat 480Gly Trp
Met Val Met Lys Gln Asn Pro Lys Arg Ile Gly Arg Thr Asp 145 150 155
160 att tcc gac ctg aac gag gac ccc gtc gtt gtc tgg cag cac cgc aac
528Ile Ser Asp Leu Asn Glu Asp Pro Val Val Val Trp Gln His Arg Asn
165 170 175 tac ctc aag gtc gtt ttc acg atg gga ttg gct gtg cct atg
ctt gtt 576Tyr Leu Lys Val Val Phe Thr Met Gly Leu Ala Val Pro Met
Leu Val 180 185 190 gct ggt ctt gga tgg ggt gac tgg ttg ggc ggc ttc
gtg tat gcc ggc 624Ala Gly Leu Gly Trp Gly Asp Trp Leu Gly Gly Phe
Val Tyr Ala Gly 195 200 205 att ctg cgt atc ttc ttc gtc cag cag gcg
act ttc tgc gtc aac tct 672Ile Leu Arg Ile Phe Phe Val Gln Gln Ala
Thr Phe Cys Val Asn Ser 210 215 220 ttg gcc cac tgg ctc ggt gac cag
ccc ttc gat gac cgc aac tca cct 720Leu Ala His Trp Leu Gly Asp Gln
Pro Phe Asp Asp Arg Asn Ser Pro 225 230 235 240 cgt gac cac gtt atc
acc gct ctc gtc acc ctt gga gag ggc tac cac 768Arg Asp His Val Ile
Thr Ala Leu Val Thr Leu Gly Glu Gly Tyr His 245 250 255 aac ttc cac
cac gag ttc ccc tcg gac tac cgt aac gcc atc gaa tgg 816Asn Phe His
His Glu Phe Pro Ser Asp Tyr Arg Asn Ala Ile Glu Trp 260 265 270 cac
cag tat gat ccc acc aag tgg tcc atc tgg gcc tgg aag cag ctt 864His
Gln Tyr Asp Pro Thr Lys Trp Ser Ile Trp Ala Trp Lys Gln Leu 275 280
285 ggt ctt gcc tac gac ctg aag aag ttc cgt gcc aac gag att gag aag
912Gly Leu Ala Tyr Asp Leu Lys Lys Phe Arg Ala Asn Glu Ile Glu Lys
290 295 300 ggt cgt gtc cag cag ctc cag aag aag ctt gac cgt aag cgt
gcc act 960Gly Arg Val Gln Gln Leu Gln Lys Lys Leu Asp Arg Lys Arg
Ala Thr 305 310 315 320 ctc gat tgg ggt act cct ctt gac cag ctc ccc
gtc atg gag tgg gac 1008Leu Asp Trp Gly Thr Pro Leu Asp Gln Leu Pro
Val Met Glu Trp Asp 325 330 335 gac tac gtc gag cag gct aag aac ggc
cgc ggt ctc gtg gct att gcc 1056Asp Tyr Val Glu Gln Ala Lys Asn Gly
Arg Gly Leu Val Ala Ile Ala 340 345 350 ggt gtt gtc cac gat gtc acg
gac ttc atc aaa gac cac ccc ggt ggc 1104Gly Val Val His Asp Val Thr
Asp Phe Ile Lys Asp His Pro Gly Gly 355 360 365 aag gcc atg atc agc
tcc ggt att ggg aag gac gcc acc gcc atg ttc 1152Lys Ala Met Ile Ser
Ser Gly Ile Gly Lys Asp Ala Thr Ala Met Phe 370 375 380 aac ggt ggt
gtc tac tac cac tcc aac gcc gca cac aac ctc ctc tct 1200Asn Gly Gly
Val Tyr Tyr His Ser Asn Ala Ala His Asn Leu Leu Ser 385 390 395 400
acc atg cgt gtt ggt gtt atc cgc ggc ggc tgt gaa gtc gaa atc tgg
1248Thr Met Arg Val Gly Val Ile Arg Gly Gly Cys Glu Val Glu Ile Trp
405 410 415 aag cgt gcc cag aag gag aac gtg gag tac gtg cgt gat ggc
tct ggc 1296Lys Arg Ala Gln Lys Glu Asn Val Glu Tyr Val Arg Asp Gly
Ser Gly 420 425 430 cag cgc gtc atc cgt gcc ggc gag cag cca acc aag
atc cca gaa ccc 1344Gln Arg Val Ile Arg Ala Gly Glu Gln Pro Thr Lys
Ile Pro Glu Pro 435 440 445 att ccc aca gcg gat gcg gcg tga 1368Ile
Pro Thr Ala Asp Ala Ala 450 455 44455PRTAspergillus nidulans 44Met
Ser Ala Pro Thr Ala Asp Ile Arg Ala Arg Ala Pro Glu Ala Lys 1 5 10
15 Lys Val His Ile Ala Asp Thr Ala Ile Asn Arg His Asn Trp Tyr Lys
20 25 30 His Val Asn Trp Leu Asn Val Phe Leu Ile Ile Gly Ile Pro
Leu Tyr 35 40 45 Gly Cys Ile Gln Ala Phe Trp Val Pro Leu Gln Leu
Lys Thr Ala Ile 50 55 60 Trp Ala Val Ile Tyr Tyr Phe Phe Thr Gly
Leu Gly Ile Thr Ala Gly 65 70 75 80 Tyr His Arg Leu Trp Ala His Cys
Ser Tyr Ser Ala Thr Leu Pro Leu 85 90 95 Arg Ile Trp Leu Ala Ala
Val Gly Gly Gly Ala Val Glu Gly Ser Ile 100 105 110 Arg Trp Trp Ala
Arg Asp His Arg Ala His His Arg Tyr Thr Asp Thr 115 120 125 Asp Lys
Asp Pro Tyr Ser Val Arg Lys Gly Leu Leu Tyr Ser His Leu 130 135 140
Gly Trp Met Val Met Lys Gln Asn Pro Lys Arg Ile Gly Arg Thr Asp 145
150 155 160 Ile Ser Asp Leu Asn Glu Asp Pro Val Val Val Trp Gln His
Arg Asn 165 170 175 Tyr Leu Lys Val Val Phe Thr Met Gly Leu Ala Val
Pro Met Leu Val 180 185 190 Ala Gly Leu Gly Trp Gly Asp Trp Leu Gly
Gly Phe Val Tyr Ala Gly 195 200 205 Ile Leu Arg Ile Phe Phe Val Gln
Gln Ala Thr Phe Cys Val Asn Ser 210 215 220 Leu Ala His Trp Leu Gly
Asp Gln Pro Phe Asp Asp Arg Asn Ser Pro 225 230 235 240 Arg Asp His
Val Ile Thr Ala Leu Val Thr Leu Gly Glu Gly Tyr His 245 250 255 Asn
Phe His His Glu Phe Pro Ser Asp Tyr Arg Asn Ala Ile Glu Trp 260 265
270 His Gln Tyr Asp Pro Thr Lys Trp Ser Ile Trp Ala Trp Lys Gln Leu
275 280 285 Gly Leu Ala Tyr Asp Leu Lys Lys Phe Arg Ala Asn Glu Ile
Glu Lys 290 295 300 Gly Arg Val Gln Gln Leu Gln Lys Lys Leu Asp Arg
Lys Arg Ala Thr 305 310 315 320 Leu Asp Trp Gly Thr Pro Leu Asp Gln
Leu Pro Val Met Glu Trp Asp 325 330 335 Asp Tyr Val Glu Gln Ala Lys
Asn Gly Arg Gly Leu Val Ala Ile Ala 340 345 350 Gly Val Val His Asp
Val Thr Asp Phe Ile Lys Asp His Pro Gly Gly 355 360 365 Lys Ala Met
Ile Ser Ser Gly Ile Gly Lys Asp Ala Thr Ala Met Phe 370 375 380 Asn
Gly Gly Val Tyr Tyr His Ser Asn Ala Ala His Asn Leu Leu Ser 385 390
395 400 Thr Met Arg Val Gly Val Ile Arg Gly Gly Cys Glu Val Glu Ile
Trp 405 410 415 Lys Arg Ala Gln Lys Glu Asn Val Glu Tyr Val Arg Asp
Gly Ser Gly 420 425 430 Gln Arg Val Ile Arg Ala Gly Glu Gln Pro Thr
Lys Ile Pro Glu Pro 435 440 445 Ile Pro Thr Ala Asp Ala Ala 450 455
451368DNAArtificial SequenceSynthetic DNA sequence encoding
Aspergillus nidulans delta-9 fatty acid desaturase protein using
codons optimized for maize and Table 1 & Table 2 sequences are
maintained 45atg agt gca cca acg gcg gac ata agg gcg cgc gcc ccg
gag gca aaa 48Met Ser Ala Pro Thr Ala Asp Ile Arg Ala Arg Ala Pro
Glu Ala Lys 1 5 10 15 aag gtt cac att gct gac act gct atc aat cgc
cat aac tgg tat aag 96Lys Val His Ile Ala Asp Thr Ala Ile Asn Arg
His Asn Trp Tyr Lys 20 25 30 cat gtg aat tgg ctg aac gtt ttt ctg
atc atc ggc atc ccg ctt tat 144His Val Asn Trp Leu Asn Val Phe Leu
Ile Ile Gly Ile Pro Leu Tyr 35 40 45 ggg tgt att caa gcg ttc tgg
gtg cca ctc cag ctc aag act gcc atc 192Gly Cys Ile Gln Ala Phe Trp
Val Pro Leu Gln Leu Lys Thr Ala Ile 50 55 60 tgg gcc gta atc tac
tac ttc ttt acc ggt ttg gga atc aca gcg ggt 240Trp Ala Val Ile Tyr
Tyr Phe Phe Thr Gly Leu Gly Ile Thr Ala Gly 65 70 75 80 tat cac aga
ttg tgg gca cac tgc tcg tac tcc gcc acc ctt cct tta 288Tyr His Arg
Leu Trp Ala His Cys Ser Tyr Ser Ala Thr Leu Pro Leu 85 90 95 cgt
ata tgg ctc gct gcc gta gga gga ggc gcc gtc gaa ggt tca atc 336Arg
Ile Trp Leu Ala Ala Val Gly Gly Gly Ala Val Glu Gly Ser Ile 100 105
110 cgt tgg tgg gct aga gac cat cgt gct cat cat aga tat acc gat aca
384Arg Trp Trp Ala Arg Asp His Arg Ala His His Arg Tyr Thr Asp Thr
115 120 125 gac aaa gac ccg tac tcc gtt
cgc aag ggg ctg cta tac tct cac ctt 432Asp Lys Asp Pro Tyr Ser Val
Arg Lys Gly Leu Leu Tyr Ser His Leu 130 135 140 ggc tgg atg gtg atg
aag cag aac cct aag cgt att ggc aga acc gat 480Gly Trp Met Val Met
Lys Gln Asn Pro Lys Arg Ile Gly Arg Thr Asp 145 150 155 160 att tcc
gac ctg aac gag gac ccc gtc gtt gtc tgg cag cac cgg aac 528Ile Ser
Asp Leu Asn Glu Asp Pro Val Val Val Trp Gln His Arg Asn 165 170 175
tac ctc aag gtc gtt ttc acg atg gga ttg gct gtg cct atg ctt gtt
576Tyr Leu Lys Val Val Phe Thr Met Gly Leu Ala Val Pro Met Leu Val
180 185 190 gct ggg ctt ggc tgg gga gac tgg ttg ggc ggc ttc gtg tat
gcc ggc 624Ala Gly Leu Gly Trp Gly Asp Trp Leu Gly Gly Phe Val Tyr
Ala Gly 195 200 205 ata ctg aga atc ttt ttc gtc cag caa gcg act ttt
tgc gtc aac tct 672Ile Leu Arg Ile Phe Phe Val Gln Gln Ala Thr Phe
Cys Val Asn Ser 210 215 220 ttg gcc cac tgg ctc gga gat cag ccg ttc
gat gac cgg aac agt cct 720Leu Ala His Trp Leu Gly Asp Gln Pro Phe
Asp Asp Arg Asn Ser Pro 225 230 235 240 agg gac cac gtt atc act gct
ctc gtc acc cta gga gag ggc tac cac 768Arg Asp His Val Ile Thr Ala
Leu Val Thr Leu Gly Glu Gly Tyr His 245 250 255 aac ttc cat cac gag
ttc ccc tcg gac tac cgg aac gcc atc gaa tgg 816Asn Phe His His Glu
Phe Pro Ser Asp Tyr Arg Asn Ala Ile Glu Trp 260 265 270 cac cag tat
gat cca acg aag tgg agc atc tgg gcc tgg aag cag ctt 864His Gln Tyr
Asp Pro Thr Lys Trp Ser Ile Trp Ala Trp Lys Gln Leu 275 280 285 ggt
tta gcc tac gac ctg aag aaa ttc aga gcc aac gag att gag aaa 912Gly
Leu Ala Tyr Asp Leu Lys Lys Phe Arg Ala Asn Glu Ile Glu Lys 290 295
300 ggg cgt gtc caa cag ctg caa aag aaa ctg gac cgt aag cgg gcg act
960Gly Arg Val Gln Gln Leu Gln Lys Lys Leu Asp Arg Lys Arg Ala Thr
305 310 315 320 ctc gat tgg gga aca cct ctg gat cag ctc ccc gtc atg
gag tgg gac 1008Leu Asp Trp Gly Thr Pro Leu Asp Gln Leu Pro Val Met
Glu Trp Asp 325 330 335 gac tac gtg gag caa gca aag aac ggt cgc ggt
ctc gtg gca ata gcg 1056Asp Tyr Val Glu Gln Ala Lys Asn Gly Arg Gly
Leu Val Ala Ile Ala 340 345 350 ggc gtg gtg cac gat gtc acg gat ttc
atc aaa gat cac ccg ggg ggc 1104Gly Val Val His Asp Val Thr Asp Phe
Ile Lys Asp His Pro Gly Gly 355 360 365 aag gcc atg atc agc tcc ggg
att ggc aag gac gca acc gcc atg ttc 1152Lys Ala Met Ile Ser Ser Gly
Ile Gly Lys Asp Ala Thr Ala Met Phe 370 375 380 aat ggg gga gtc tac
tac cac agc aac gca gca cac aat ctc ttg tca 1200Asn Gly Gly Val Tyr
Tyr His Ser Asn Ala Ala His Asn Leu Leu Ser 385 390 395 400 aca atg
agg gtg ggt gtt att agg ggc ggc tgt gaa gtc gaa atc tgg 1248Thr Met
Arg Val Gly Val Ile Arg Gly Gly Cys Glu Val Glu Ile Trp 405 410 415
aag agg gcg caa aag gag aat gtg gag tac gtg cga gat ggc tct ggt
1296Lys Arg Ala Gln Lys Glu Asn Val Glu Tyr Val Arg Asp Gly Ser Gly
420 425 430 caa cgc gtg atc aga gcg ggc gag cag cca acc aag ata cca
gaa ccg 1344Gln Arg Val Ile Arg Ala Gly Glu Gln Pro Thr Lys Ile Pro
Glu Pro 435 440 445 att ccc aca gcg gat gcg gcg tag 1368Ile Pro Thr
Ala Asp Ala Ala 450 455 46455PRTArtificial SequenceSynthetic
Construct 46Met Ser Ala Pro Thr Ala Asp Ile Arg Ala Arg Ala Pro Glu
Ala Lys 1 5 10 15 Lys Val His Ile Ala Asp Thr Ala Ile Asn Arg His
Asn Trp Tyr Lys 20 25 30 His Val Asn Trp Leu Asn Val Phe Leu Ile
Ile Gly Ile Pro Leu Tyr 35 40 45 Gly Cys Ile Gln Ala Phe Trp Val
Pro Leu Gln Leu Lys Thr Ala Ile 50 55 60 Trp Ala Val Ile Tyr Tyr
Phe Phe Thr Gly Leu Gly Ile Thr Ala Gly 65 70 75 80 Tyr His Arg Leu
Trp Ala His Cys Ser Tyr Ser Ala Thr Leu Pro Leu 85 90 95 Arg Ile
Trp Leu Ala Ala Val Gly Gly Gly Ala Val Glu Gly Ser Ile 100 105 110
Arg Trp Trp Ala Arg Asp His Arg Ala His His Arg Tyr Thr Asp Thr 115
120 125 Asp Lys Asp Pro Tyr Ser Val Arg Lys Gly Leu Leu Tyr Ser His
Leu 130 135 140 Gly Trp Met Val Met Lys Gln Asn Pro Lys Arg Ile Gly
Arg Thr Asp 145 150 155 160 Ile Ser Asp Leu Asn Glu Asp Pro Val Val
Val Trp Gln His Arg Asn 165 170 175 Tyr Leu Lys Val Val Phe Thr Met
Gly Leu Ala Val Pro Met Leu Val 180 185 190 Ala Gly Leu Gly Trp Gly
Asp Trp Leu Gly Gly Phe Val Tyr Ala Gly 195 200 205 Ile Leu Arg Ile
Phe Phe Val Gln Gln Ala Thr Phe Cys Val Asn Ser 210 215 220 Leu Ala
His Trp Leu Gly Asp Gln Pro Phe Asp Asp Arg Asn Ser Pro 225 230 235
240 Arg Asp His Val Ile Thr Ala Leu Val Thr Leu Gly Glu Gly Tyr His
245 250 255 Asn Phe His His Glu Phe Pro Ser Asp Tyr Arg Asn Ala Ile
Glu Trp 260 265 270 His Gln Tyr Asp Pro Thr Lys Trp Ser Ile Trp Ala
Trp Lys Gln Leu 275 280 285 Gly Leu Ala Tyr Asp Leu Lys Lys Phe Arg
Ala Asn Glu Ile Glu Lys 290 295 300 Gly Arg Val Gln Gln Leu Gln Lys
Lys Leu Asp Arg Lys Arg Ala Thr 305 310 315 320 Leu Asp Trp Gly Thr
Pro Leu Asp Gln Leu Pro Val Met Glu Trp Asp 325 330 335 Asp Tyr Val
Glu Gln Ala Lys Asn Gly Arg Gly Leu Val Ala Ile Ala 340 345 350 Gly
Val Val His Asp Val Thr Asp Phe Ile Lys Asp His Pro Gly Gly 355 360
365 Lys Ala Met Ile Ser Ser Gly Ile Gly Lys Asp Ala Thr Ala Met Phe
370 375 380 Asn Gly Gly Val Tyr Tyr His Ser Asn Ala Ala His Asn Leu
Leu Ser 385 390 395 400 Thr Met Arg Val Gly Val Ile Arg Gly Gly Cys
Glu Val Glu Ile Trp 405 410 415 Lys Arg Ala Gln Lys Glu Asn Val Glu
Tyr Val Arg Asp Gly Ser Gly 420 425 430 Gln Arg Val Ile Arg Ala Gly
Glu Gln Pro Thr Lys Ile Pro Glu Pro 435 440 445 Ile Pro Thr Ala Asp
Ala Ala 450 455 471368DNAArtificial SequenceSynthetic DNA sequence
in accordance with the invention encoding Aspergillus nidulans
delta-9 fatty acid desaturase protein using codons optimized for
maize and with sequences identified in Table 2 removed and Table 1
sequences are maintained 47atg agt gca cca acg gcg gac ata agg gcg
cgc gcc ccg gag gca aaa 48Met Ser Ala Pro Thr Ala Asp Ile Arg Ala
Arg Ala Pro Glu Ala Lys 1 5 10 15 aag gtt cac att gct gac act gct
atc aat cgc cat aac tgg tat aag 96Lys Val His Ile Ala Asp Thr Ala
Ile Asn Arg His Asn Trp Tyr Lys 20 25 30 cat gtg aat tgg ctg aac
gtt ttt ctg atc atc ggc atc ccg ctt tat 144His Val Asn Trp Leu Asn
Val Phe Leu Ile Ile Gly Ile Pro Leu Tyr 35 40 45 ggg tgt att caa
gcg ttc tgg gtg cca ctc cag ctc aag act gcc atc 192Gly Cys Ile Gln
Ala Phe Trp Val Pro Leu Gln Leu Lys Thr Ala Ile 50 55 60 tgg gcc
gta atc tac tac ttc ttt acc ggt ttg gga atc aca gcg ggt 240Trp Ala
Val Ile Tyr Tyr Phe Phe Thr Gly Leu Gly Ile Thr Ala Gly 65 70 75 80
tat cac aga ttg tgg gca cac tgc tcg tac tcc gcc acc ctt cct tta
288Tyr His Arg Leu Trp Ala His Cys Ser Tyr Ser Ala Thr Leu Pro Leu
85 90 95 cgt ata tgg ctc gct gcc gta gga gga ggc gcc gtc gaa ggt
tca atc 336Arg Ile Trp Leu Ala Ala Val Gly Gly Gly Ala Val Glu Gly
Ser Ile 100 105 110 cgt tgg tgg gct aga gac cat cgt gct cat cat aga
tat acc gat aca 384Arg Trp Trp Ala Arg Asp His Arg Ala His His Arg
Tyr Thr Asp Thr 115 120 125 gac aaa gac ccg tac tcc gtt cgc aag ggg
ctg cta tac tct cac ctt 432Asp Lys Asp Pro Tyr Ser Val Arg Lys Gly
Leu Leu Tyr Ser His Leu 130 135 140 ggc tgg atg gtg atg aag cag aac
cct aag cgt att ggc aga acc gat 480Gly Trp Met Val Met Lys Gln Asn
Pro Lys Arg Ile Gly Arg Thr Asp 145 150 155 160 att agc gac ctg aac
gag gac ccc gtc gtt gtc tgg cag cac cgg aac 528Ile Ser Asp Leu Asn
Glu Asp Pro Val Val Val Trp Gln His Arg Asn 165 170 175 tac ctc aag
gtc gtt ttc acg atg gga ttg gct gtg cct atg ctt gtt 576Tyr Leu Lys
Val Val Phe Thr Met Gly Leu Ala Val Pro Met Leu Val 180 185 190 gct
ggg ctt ggc tgg gga gac tgg ttg ggc ggc ttc gtg tat gcc ggc 624Ala
Gly Leu Gly Trp Gly Asp Trp Leu Gly Gly Phe Val Tyr Ala Gly 195 200
205 ata ctg aga atc ttt ttc gtc cag caa gcg act ttt tgc gtc aac tct
672Ile Leu Arg Ile Phe Phe Val Gln Gln Ala Thr Phe Cys Val Asn Ser
210 215 220 ttg gcc cac tgg ctc gga gat cag ccg ttc gat gac cgg aac
agt cct 720Leu Ala His Trp Leu Gly Asp Gln Pro Phe Asp Asp Arg Asn
Ser Pro 225 230 235 240 agg gac cac gtt atc act gct ctc gtc acc cta
gga gag ggc tac cac 768Arg Asp His Val Ile Thr Ala Leu Val Thr Leu
Gly Glu Gly Tyr His 245 250 255 aac ttc cat cac gag ttc ccc tcg gac
tac cgg aac gcc atc gaa tgg 816Asn Phe His His Glu Phe Pro Ser Asp
Tyr Arg Asn Ala Ile Glu Trp 260 265 270 cac cag tat gat cca acg aag
tgg agc atc tgg gcc tgg aag cag ctt 864His Gln Tyr Asp Pro Thr Lys
Trp Ser Ile Trp Ala Trp Lys Gln Leu 275 280 285 ggt tta gcc tac gac
ctg aag aaa ttc aga gcc aac gag att gag aaa 912Gly Leu Ala Tyr Asp
Leu Lys Lys Phe Arg Ala Asn Glu Ile Glu Lys 290 295 300 ggg cgt gtc
caa cag ctg caa aag aaa ctg gac cgt aag cgg gcg act 960Gly Arg Val
Gln Gln Leu Gln Lys Lys Leu Asp Arg Lys Arg Ala Thr 305 310 315 320
ctc gat tgg gga aca cct ctg gat cag ctc ccc gtc atg gag tgg gac
1008Leu Asp Trp Gly Thr Pro Leu Asp Gln Leu Pro Val Met Glu Trp Asp
325 330 335 gac tac gtg gag caa gca aag aac ggt cgc ggt ctc gtg gca
ata gcg 1056Asp Tyr Val Glu Gln Ala Lys Asn Gly Arg Gly Leu Val Ala
Ile Ala 340 345 350 ggc gtg gtg cac gat gtc acg gat ttc atc aaa gat
cac ccg ggg ggc 1104Gly Val Val His Asp Val Thr Asp Phe Ile Lys Asp
His Pro Gly Gly 355 360 365 aag gcc atg atc agc tcc ggg att ggc aag
gac gca acc gcc atg ttc 1152Lys Ala Met Ile Ser Ser Gly Ile Gly Lys
Asp Ala Thr Ala Met Phe 370 375 380 aat ggg gga gtc tac tac cac agc
aac gca gca cac aat ctc ttg tca 1200Asn Gly Gly Val Tyr Tyr His Ser
Asn Ala Ala His Asn Leu Leu Ser 385 390 395 400 aca atg agg gtg ggt
gtt att agg ggc ggc tgt gaa gtc gaa atc tgg 1248Thr Met Arg Val Gly
Val Ile Arg Gly Gly Cys Glu Val Glu Ile Trp 405 410 415 aag agg gcg
caa aag gag aat gtg gag tac gtg cga gat ggc tct ggt 1296Lys Arg Ala
Gln Lys Glu Asn Val Glu Tyr Val Arg Asp Gly Ser Gly 420 425 430 caa
cgc gtg atc aga gcg ggc gag cag cca acc aag ata cca gaa ccg 1344Gln
Arg Val Ile Arg Ala Gly Glu Gln Pro Thr Lys Ile Pro Glu Pro 435 440
445 att ccc aca gcg gat gcg gcg tag 1368Ile Pro Thr Ala Asp Ala Ala
450 455 48455PRTArtificial SequenceSynthetic Construct 48Met Ser
Ala Pro Thr Ala Asp Ile Arg Ala Arg Ala Pro Glu Ala Lys 1 5 10 15
Lys Val His Ile Ala Asp Thr Ala Ile Asn Arg His Asn Trp Tyr Lys 20
25 30 His Val Asn Trp Leu Asn Val Phe Leu Ile Ile Gly Ile Pro Leu
Tyr 35 40 45 Gly Cys Ile Gln Ala Phe Trp Val Pro Leu Gln Leu Lys
Thr Ala Ile 50 55 60 Trp Ala Val Ile Tyr Tyr Phe Phe Thr Gly Leu
Gly Ile Thr Ala Gly 65 70 75 80 Tyr His Arg Leu Trp Ala His Cys Ser
Tyr Ser Ala Thr Leu Pro Leu 85 90 95 Arg Ile Trp Leu Ala Ala Val
Gly Gly Gly Ala Val Glu Gly Ser Ile 100 105 110 Arg Trp Trp Ala Arg
Asp His Arg Ala His His Arg Tyr Thr Asp Thr 115 120 125 Asp Lys Asp
Pro Tyr Ser Val Arg Lys Gly Leu Leu Tyr Ser His Leu 130 135 140 Gly
Trp Met Val Met Lys Gln Asn Pro Lys Arg Ile Gly Arg Thr Asp 145 150
155 160 Ile Ser Asp Leu Asn Glu Asp Pro Val Val Val Trp Gln His Arg
Asn 165 170 175 Tyr Leu Lys Val Val Phe Thr Met Gly Leu Ala Val Pro
Met Leu Val 180 185 190 Ala Gly Leu Gly Trp Gly Asp Trp Leu Gly Gly
Phe Val Tyr Ala Gly 195 200 205 Ile Leu Arg Ile Phe Phe Val Gln Gln
Ala Thr Phe Cys Val Asn Ser 210 215 220 Leu Ala His Trp Leu Gly Asp
Gln Pro Phe Asp Asp Arg Asn Ser Pro 225 230 235 240 Arg Asp His Val
Ile Thr Ala Leu Val Thr Leu Gly Glu Gly Tyr His 245 250 255 Asn Phe
His His Glu Phe Pro Ser Asp Tyr Arg Asn Ala Ile Glu Trp 260 265 270
His Gln Tyr Asp Pro Thr Lys Trp Ser Ile Trp Ala Trp Lys Gln Leu 275
280 285 Gly Leu Ala Tyr Asp Leu Lys Lys Phe Arg Ala Asn Glu Ile Glu
Lys 290 295 300 Gly Arg Val Gln Gln Leu Gln Lys Lys Leu Asp Arg Lys
Arg Ala Thr 305 310 315 320 Leu Asp Trp Gly Thr Pro Leu Asp Gln Leu
Pro Val Met Glu Trp Asp 325 330 335 Asp Tyr Val Glu Gln Ala Lys Asn
Gly Arg Gly Leu Val Ala Ile Ala 340 345 350 Gly Val Val His Asp Val
Thr Asp Phe Ile Lys Asp His Pro Gly Gly 355 360 365 Lys Ala Met Ile
Ser Ser Gly Ile Gly Lys Asp Ala Thr Ala Met Phe 370 375 380 Asn Gly
Gly Val Tyr Tyr His Ser Asn Ala Ala His Asn Leu Leu Ser 385 390 395
400 Thr Met Arg Val Gly Val Ile Arg Gly Gly Cys Glu Val Glu Ile Trp
405 410 415 Lys Arg Ala Gln Lys Glu Asn Val Glu Tyr Val Arg Asp Gly
Ser Gly 420 425 430 Gln Arg Val Ile Arg Ala Gly
Glu Gln Pro Thr Lys Ile Pro Glu Pro 435 440 445 Ile Pro Thr Ala Asp
Ala Ala 450 455 49798DNAXerophyta viscosaCDS(1)..(798)Native DNA
sequence encoding Xerophyta viscosa SAP1 protein 49atg agg aac gag
ggt ttt ctg aaa atg aag acc gac gtt gga gtc gcc 48Met Arg Asn Glu
Gly Phe Leu Lys Met Lys Thr Asp Val Gly Val Ala 1 5 10 15 gac gag
gtg atc tcc gga gat ctc aag cag ctt ggt gac gct gca aag 96Asp Glu
Val Ile Ser Gly Asp Leu Lys Gln Leu Gly Asp Ala Ala Lys 20 25 30
cgg cta gct aaa cat gcg atc aag ctc ggc gcc agc ttc ggg gtt ggc
144Arg Leu Ala Lys His Ala Ile Lys Leu Gly Ala Ser Phe Gly Val Gly
35 40 45 tct acc ata gtc cag gct att gct tcg atc gct gct atc tat
ttg ttg 192Ser Thr Ile Val Gln Ala Ile Ala Ser Ile Ala Ala Ile Tyr
Leu Leu 50 55 60 ata ttg gac cgg aca aac tgg cgt aca aat atc ttg
aca tca ctt cta 240Ile Leu Asp Arg Thr Asn Trp Arg Thr Asn Ile Leu
Thr Ser Leu Leu 65 70 75 80 att cca tat gtt tac ttg agt ctt cct tca
gtg ata ttc aac cta ttc 288Ile Pro Tyr Val Tyr Leu Ser Leu Pro Ser
Val Ile Phe Asn Leu Phe 85 90 95 agg ggc gac ctg ggc aga tgg ctt
tca ttc att ggc gta gta atg aag 336Arg Gly Asp Leu Gly Arg Trp Leu
Ser Phe Ile Gly Val Val Met Lys 100 105 110 ctc ttc ttc cac cga cac
ttc cca gtt acc ttg gaa ctg ctt gtg tct 384Leu Phe Phe His Arg His
Phe Pro Val Thr Leu Glu Leu Leu Val Ser 115 120 125 ctc att ctc ctg
att gtg gtt tcc ccc act ttc att gcc cac aca atc 432Leu Ile Leu Leu
Ile Val Val Ser Pro Thr Phe Ile Ala His Thr Ile 130 135 140 aga ggc
agt ctc att gga gtc ttc atc ttc ctt gtc atc gcc tgc tac 480Arg Gly
Ser Leu Ile Gly Val Phe Ile Phe Leu Val Ile Ala Cys Tyr 145 150 155
160 ctc ctc caa gag cac att aga tca gct ggt ggc ttc aaa aac gcg ttc
528Leu Leu Gln Glu His Ile Arg Ser Ala Gly Gly Phe Lys Asn Ala Phe
165 170 175 aca aag agc aat ggg att tca aac agc gtc ggg atc atc att
cta ctg 576Thr Lys Ser Asn Gly Ile Ser Asn Ser Val Gly Ile Ile Ile
Leu Leu 180 185 190 atc cac ccg atc tgg agc ttg gtg gtg tat ttc ctc
tac acg tct ttg 624Ile His Pro Ile Trp Ser Leu Val Val Tyr Phe Leu
Tyr Thr Ser Leu 195 200 205 ctg caa ctt ctt gca tac tct cct tcc cct
tgt tgt tgc ata tta tac 672Leu Gln Leu Leu Ala Tyr Ser Pro Ser Pro
Cys Cys Cys Ile Leu Tyr 210 215 220 aat aag tgg ttt aat ttc atg cat
gtt tgt aaa tgt gta agc ctt cat 720Asn Lys Trp Phe Asn Phe Met His
Val Cys Lys Cys Val Ser Leu His 225 230 235 240 atg tat tct cag tca
att ggg tca tgc gtg tcc ata ttt ttc gtg cag 768Met Tyr Ser Gln Ser
Ile Gly Ser Cys Val Ser Ile Phe Phe Val Gln 245 250 255 ttt gta ttc
atc tat gaa gct gaa ttt taa 798Phe Val Phe Ile Tyr Glu Ala Glu Phe
260 265 50265PRTXerophyta viscosa 50Met Arg Asn Glu Gly Phe Leu Lys
Met Lys Thr Asp Val Gly Val Ala 1 5 10 15 Asp Glu Val Ile Ser Gly
Asp Leu Lys Gln Leu Gly Asp Ala Ala Lys 20 25 30 Arg Leu Ala Lys
His Ala Ile Lys Leu Gly Ala Ser Phe Gly Val Gly 35 40 45 Ser Thr
Ile Val Gln Ala Ile Ala Ser Ile Ala Ala Ile Tyr Leu Leu 50 55 60
Ile Leu Asp Arg Thr Asn Trp Arg Thr Asn Ile Leu Thr Ser Leu Leu 65
70 75 80 Ile Pro Tyr Val Tyr Leu Ser Leu Pro Ser Val Ile Phe Asn
Leu Phe 85 90 95 Arg Gly Asp Leu Gly Arg Trp Leu Ser Phe Ile Gly
Val Val Met Lys 100 105 110 Leu Phe Phe His Arg His Phe Pro Val Thr
Leu Glu Leu Leu Val Ser 115 120 125 Leu Ile Leu Leu Ile Val Val Ser
Pro Thr Phe Ile Ala His Thr Ile 130 135 140 Arg Gly Ser Leu Ile Gly
Val Phe Ile Phe Leu Val Ile Ala Cys Tyr 145 150 155 160 Leu Leu Gln
Glu His Ile Arg Ser Ala Gly Gly Phe Lys Asn Ala Phe 165 170 175 Thr
Lys Ser Asn Gly Ile Ser Asn Ser Val Gly Ile Ile Ile Leu Leu 180 185
190 Ile His Pro Ile Trp Ser Leu Val Val Tyr Phe Leu Tyr Thr Ser Leu
195 200 205 Leu Gln Leu Leu Ala Tyr Ser Pro Ser Pro Cys Cys Cys Ile
Leu Tyr 210 215 220 Asn Lys Trp Phe Asn Phe Met His Val Cys Lys Cys
Val Ser Leu His 225 230 235 240 Met Tyr Ser Gln Ser Ile Gly Ser Cys
Val Ser Ile Phe Phe Val Gln 245 250 255 Phe Val Phe Ile Tyr Glu Ala
Glu Phe 260 265 51798DNAArtificial SequenceSynthetic DNA sequence
encoding Xerophyta viscosa SAP1 protein using codons optimized for
maize and Table 1 & Table 2 sequences are maintained 51atg aga
aac gaa ggt ttt ctg aag atg aaa acg gac gtt ggg gtt gct 48Met Arg
Asn Glu Gly Phe Leu Lys Met Lys Thr Asp Val Gly Val Ala 1 5 10 15
gac gaa gtc atc agc ggt gat ttg aag cag ttg ggt gat gct gcc aaa
96Asp Glu Val Ile Ser Gly Asp Leu Lys Gln Leu Gly Asp Ala Ala Lys
20 25 30 cgc ctt gct aag cac gct atc aaa ctg gga gcc agc ttt ggt
gtt ggt 144Arg Leu Ala Lys His Ala Ile Lys Leu Gly Ala Ser Phe Gly
Val Gly 35 40 45 tca act atc gtt caa gcc atc gca tca ata gca gcc
atc tat ctt ctg 192Ser Thr Ile Val Gln Ala Ile Ala Ser Ile Ala Ala
Ile Tyr Leu Leu 50 55 60 att ctc gat agg acc aac tgg agg acc aac
atc ttg acg tcc ctc ctc 240Ile Leu Asp Arg Thr Asn Trp Arg Thr Asn
Ile Leu Thr Ser Leu Leu 65 70 75 80 att ccc tac gtg tat ctg tcc ctc
ccg agc gtc atc ttc aat ctc ttt 288Ile Pro Tyr Val Tyr Leu Ser Leu
Pro Ser Val Ile Phe Asn Leu Phe 85 90 95 cgt ggg gac ctc ggg aga
tgg ctg tca ttc ata ggc gtt gtg atg aag 336Arg Gly Asp Leu Gly Arg
Trp Leu Ser Phe Ile Gly Val Val Met Lys 100 105 110 ctg ttc ttt cat
agg cac ttt cct gtt act ttg gag ctg ctt gtg agc 384Leu Phe Phe His
Arg His Phe Pro Val Thr Leu Glu Leu Leu Val Ser 115 120 125 ctc att
ctt ttg att gtc gtg tca cct acc ttc ata gct cat aca att 432Leu Ile
Leu Leu Ile Val Val Ser Pro Thr Phe Ile Ala His Thr Ile 130 135 140
cgt gga tct ttg att ggg gtg ttc atc ttc ttg gtg ata gca tgt tat
480Arg Gly Ser Leu Ile Gly Val Phe Ile Phe Leu Val Ile Ala Cys Tyr
145 150 155 160 ctg ctt caa gag cac att aga tca gct ggt ggc ttc aag
aac gcc ttt 528Leu Leu Gln Glu His Ile Arg Ser Ala Gly Gly Phe Lys
Asn Ala Phe 165 170 175 aca aag tct aat gga atc tcc aac agc gtg ggc
atc atc atc ctt ctg 576Thr Lys Ser Asn Gly Ile Ser Asn Ser Val Gly
Ile Ile Ile Leu Leu 180 185 190 atc cac ccg att tgg tct ctc gtc gtc
tac ttc ctc tac act tca ctt 624Ile His Pro Ile Trp Ser Leu Val Val
Tyr Phe Leu Tyr Thr Ser Leu 195 200 205 ctc cag ctt ttg gcc tac tca
cca tcc ccg tgc tgc tgc ata tta tac 672Leu Gln Leu Leu Ala Tyr Ser
Pro Ser Pro Cys Cys Cys Ile Leu Tyr 210 215 220 aac aag tgg ttc aac
ttc atg cat gtt tgc aag tgc gtc tct ttg cac 720Asn Lys Trp Phe Asn
Phe Met His Val Cys Lys Cys Val Ser Leu His 225 230 235 240 atg tac
tct cag tcc ata ggc tca tgt gtt tca ata ttt ttc gtc cag 768Met Tyr
Ser Gln Ser Ile Gly Ser Cys Val Ser Ile Phe Phe Val Gln 245 250 255
ttc gtg ttc atc tat gag gct gag ttt taa 798Phe Val Phe Ile Tyr Glu
Ala Glu Phe 260 265 52265PRTArtificial SequenceSynthetic Construct
52Met Arg Asn Glu Gly Phe Leu Lys Met Lys Thr Asp Val Gly Val Ala 1
5 10 15 Asp Glu Val Ile Ser Gly Asp Leu Lys Gln Leu Gly Asp Ala Ala
Lys 20 25 30 Arg Leu Ala Lys His Ala Ile Lys Leu Gly Ala Ser Phe
Gly Val Gly 35 40 45 Ser Thr Ile Val Gln Ala Ile Ala Ser Ile Ala
Ala Ile Tyr Leu Leu 50 55 60 Ile Leu Asp Arg Thr Asn Trp Arg Thr
Asn Ile Leu Thr Ser Leu Leu 65 70 75 80 Ile Pro Tyr Val Tyr Leu Ser
Leu Pro Ser Val Ile Phe Asn Leu Phe 85 90 95 Arg Gly Asp Leu Gly
Arg Trp Leu Ser Phe Ile Gly Val Val Met Lys 100 105 110 Leu Phe Phe
His Arg His Phe Pro Val Thr Leu Glu Leu Leu Val Ser 115 120 125 Leu
Ile Leu Leu Ile Val Val Ser Pro Thr Phe Ile Ala His Thr Ile 130 135
140 Arg Gly Ser Leu Ile Gly Val Phe Ile Phe Leu Val Ile Ala Cys Tyr
145 150 155 160 Leu Leu Gln Glu His Ile Arg Ser Ala Gly Gly Phe Lys
Asn Ala Phe 165 170 175 Thr Lys Ser Asn Gly Ile Ser Asn Ser Val Gly
Ile Ile Ile Leu Leu 180 185 190 Ile His Pro Ile Trp Ser Leu Val Val
Tyr Phe Leu Tyr Thr Ser Leu 195 200 205 Leu Gln Leu Leu Ala Tyr Ser
Pro Ser Pro Cys Cys Cys Ile Leu Tyr 210 215 220 Asn Lys Trp Phe Asn
Phe Met His Val Cys Lys Cys Val Ser Leu His 225 230 235 240 Met Tyr
Ser Gln Ser Ile Gly Ser Cys Val Ser Ile Phe Phe Val Gln 245 250 255
Phe Val Phe Ile Tyr Glu Ala Glu Phe 260 265 53798DNAArtificial
SequenceSynthetic DNA sequence in accordance with the invention
encoding Xerophyta viscosa SAP1 protein using codons optimized for
maize and with sequences identified in Table 2 removed and Table 1
sequences are maintained 53atg aga aac gaa ggt ttt ctg aag atg aaa
acg gac gtt ggg gtt gct 48Met Arg Asn Glu Gly Phe Leu Lys Met Lys
Thr Asp Val Gly Val Ala 1 5 10 15 gac gaa gtc atc agc ggt gat ttg
aag cag ttg ggt gat gct gcc aaa 96Asp Glu Val Ile Ser Gly Asp Leu
Lys Gln Leu Gly Asp Ala Ala Lys 20 25 30 cgc ctt gct aag cac gct
atc aaa ctg gga gcc agc ttt ggt gtt ggt 144Arg Leu Ala Lys His Ala
Ile Lys Leu Gly Ala Ser Phe Gly Val Gly 35 40 45 tca act atc gtt
caa gcc atc gca tca ata gca gcc atc tat ctt ctg 192Ser Thr Ile Val
Gln Ala Ile Ala Ser Ile Ala Ala Ile Tyr Leu Leu 50 55 60 att ctc
gat agg acc aac tgg agg acc aac atc ttg acg tcc ctc ctc 240Ile Leu
Asp Arg Thr Asn Trp Arg Thr Asn Ile Leu Thr Ser Leu Leu 65 70 75 80
att ccc tac gtg tat ctg tcc ctc ccg agc gtc atc ttc aat ctc ttt
288Ile Pro Tyr Val Tyr Leu Ser Leu Pro Ser Val Ile Phe Asn Leu Phe
85 90 95 cgt ggg gac ctc ggg aga tgg ctg tca ttc ata ggc gtt gtg
atg aag 336Arg Gly Asp Leu Gly Arg Trp Leu Ser Phe Ile Gly Val Val
Met Lys 100 105 110 ctg ttc ttt cat agg cac ttt cct gtt act ttg gag
ctg ctt gtg agc 384Leu Phe Phe His Arg His Phe Pro Val Thr Leu Glu
Leu Leu Val Ser 115 120 125 ctc att ctt ttg att gtc gtg tct cct acc
ttc ata gct cat aca att 432Leu Ile Leu Leu Ile Val Val Ser Pro Thr
Phe Ile Ala His Thr Ile 130 135 140 cgt gga tct ttg att ggg gtg ttc
atc ttc ttg gtg ata gca tgt tat 480Arg Gly Ser Leu Ile Gly Val Phe
Ile Phe Leu Val Ile Ala Cys Tyr 145 150 155 160 ctg ctt caa gag cac
att aga tca gct ggt ggc ttc aag aac gcc ttt 528Leu Leu Gln Glu His
Ile Arg Ser Ala Gly Gly Phe Lys Asn Ala Phe 165 170 175 aca aag tct
aat gga atc tcc aac agc gtg ggc atc atc atc ctt ctg 576Thr Lys Ser
Asn Gly Ile Ser Asn Ser Val Gly Ile Ile Ile Leu Leu 180 185 190 atc
cac ccg att tgg tct ctc gtc gtc tac ttc ctc tac act tca ctt 624Ile
His Pro Ile Trp Ser Leu Val Val Tyr Phe Leu Tyr Thr Ser Leu 195 200
205 ctc cag ctt ttg gcc tac tca cca tcc cca tgc tgc tgt att ctt tac
672Leu Gln Leu Leu Ala Tyr Ser Pro Ser Pro Cys Cys Cys Ile Leu Tyr
210 215 220 aac aaa tgg ttc aac ttc atg cac gtg tgc aag tgc gtc tct
ttg cac 720Asn Lys Trp Phe Asn Phe Met His Val Cys Lys Cys Val Ser
Leu His 225 230 235 240 atg tac tct cag tcc att ggc tca tgt gtt tca
atc ttc ttt gtc cag 768Met Tyr Ser Gln Ser Ile Gly Ser Cys Val Ser
Ile Phe Phe Val Gln 245 250 255 ttc gtg ttc atc tat gag gct gag ttt
taa 798Phe Val Phe Ile Tyr Glu Ala Glu Phe 260 265
54265PRTArtificial SequenceSynthetic Construct 54Met Arg Asn Glu
Gly Phe Leu Lys Met Lys Thr Asp Val Gly Val Ala 1 5 10 15 Asp Glu
Val Ile Ser Gly Asp Leu Lys Gln Leu Gly Asp Ala Ala Lys 20 25 30
Arg Leu Ala Lys His Ala Ile Lys Leu Gly Ala Ser Phe Gly Val Gly 35
40 45 Ser Thr Ile Val Gln Ala Ile Ala Ser Ile Ala Ala Ile Tyr Leu
Leu 50 55 60 Ile Leu Asp Arg Thr Asn Trp Arg Thr Asn Ile Leu Thr
Ser Leu Leu 65 70 75 80 Ile Pro Tyr Val Tyr Leu Ser Leu Pro Ser Val
Ile Phe Asn Leu Phe 85 90 95 Arg Gly Asp Leu Gly Arg Trp Leu Ser
Phe Ile Gly Val Val Met Lys 100 105 110 Leu Phe Phe His Arg His Phe
Pro Val Thr Leu Glu Leu Leu Val Ser 115 120 125 Leu Ile Leu Leu Ile
Val Val Ser Pro Thr Phe Ile Ala His Thr Ile 130 135 140 Arg Gly Ser
Leu Ile Gly Val Phe Ile Phe Leu Val Ile Ala Cys Tyr 145 150 155 160
Leu Leu Gln Glu His Ile Arg Ser Ala Gly Gly Phe Lys Asn Ala Phe 165
170 175 Thr Lys Ser Asn Gly Ile Ser Asn Ser Val Gly Ile Ile Ile Leu
Leu 180 185 190 Ile His Pro Ile Trp Ser Leu Val Val Tyr Phe Leu Tyr
Thr Ser Leu 195 200 205 Leu Gln Leu Leu Ala Tyr Ser Pro Ser Pro Cys
Cys Cys Ile Leu Tyr 210 215 220 Asn Lys Trp Phe Asn Phe Met His Val
Cys Lys Cys Val Ser Leu His 225 230 235 240 Met Tyr Ser Gln Ser Ile
Gly Ser Cys Val Ser Ile Phe Phe Val Gln 245 250 255 Phe Val Phe Ile
Tyr Glu Ala Glu Phe 260 265 55717DNAAequorea
victoriaCDS(1)..(717)Native DNA sequence encoding Aequorea
victoria
GFP1 protein 55atg agt aaa gga gaa gaa ctt ttc act gga gtg gtc cca
gtt ctt gtt 48Met Ser Lys Gly Glu Glu Leu Phe Thr Gly Val Val Pro
Val Leu Val 1 5 10 15 gaa tta gat ggc gat gtt aat ggg caa aaa ttc
tct gtc agt gga gag 96Glu Leu Asp Gly Asp Val Asn Gly Gln Lys Phe
Ser Val Ser Gly Glu 20 25 30 ggt gaa ggt gat gca aca tac gga aaa
ctt acc ctt aat ttt att tgc 144Gly Glu Gly Asp Ala Thr Tyr Gly Lys
Leu Thr Leu Asn Phe Ile Cys 35 40 45 act act ggg aag cta cct gtt
cca tgg cca aca ctt gtc act act ttc 192Thr Thr Gly Lys Leu Pro Val
Pro Trp Pro Thr Leu Val Thr Thr Phe 50 55 60 tct tat ggt gtt caa
tgc ttc tca aga tac cca gat cat atg aaa cag 240Ser Tyr Gly Val Gln
Cys Phe Ser Arg Tyr Pro Asp His Met Lys Gln 65 70 75 80 cat gac ttt
ttc aag agt gcc atg ccc gaa ggt tat gta cag gaa aga 288His Asp Phe
Phe Lys Ser Ala Met Pro Glu Gly Tyr Val Gln Glu Arg 85 90 95 act
ata ttt tac aaa gat gac ggg aac tac aag aca cgt gct gaa gtc 336Thr
Ile Phe Tyr Lys Asp Asp Gly Asn Tyr Lys Thr Arg Ala Glu Val 100 105
110 aag ttt gaa ggt gat acc ctt gtt aat aga atc gag tta aaa ggt att
384Lys Phe Glu Gly Asp Thr Leu Val Asn Arg Ile Glu Leu Lys Gly Ile
115 120 125 gat ttt aaa gaa gat gga aac att ctt gga cac aaa atg gaa
tac aac 432Asp Phe Lys Glu Asp Gly Asn Ile Leu Gly His Lys Met Glu
Tyr Asn 130 135 140 tat aac tca cat aat gta tac atc atg gga gac aaa
cca aag aat ggc 480Tyr Asn Ser His Asn Val Tyr Ile Met Gly Asp Lys
Pro Lys Asn Gly 145 150 155 160 atc aaa gtt aac ttc aaa att aga cac
aac att aaa gat gga agc gtt 528Ile Lys Val Asn Phe Lys Ile Arg His
Asn Ile Lys Asp Gly Ser Val 165 170 175 caa tta gca gac cat tat caa
caa aat act cca att ggc gat ggc cct 576Gln Leu Ala Asp His Tyr Gln
Gln Asn Thr Pro Ile Gly Asp Gly Pro 180 185 190 gtc ctt tta cca gac
aac cat tac ctg tcc aca caa tct gcc ctt tcc 624Val Leu Leu Pro Asp
Asn His Tyr Leu Ser Thr Gln Ser Ala Leu Ser 195 200 205 aaa gat ccc
aac gaa aag aga gat cac atg atc ctt ctt gag ttt gta 672Lys Asp Pro
Asn Glu Lys Arg Asp His Met Ile Leu Leu Glu Phe Val 210 215 220 aca
gct gct agg att aca cat ggc atg gat gaa cta tac aaa taa 717Thr Ala
Ala Arg Ile Thr His Gly Met Asp Glu Leu Tyr Lys 225 230 235
56238PRTAequorea victoria 56Met Ser Lys Gly Glu Glu Leu Phe Thr Gly
Val Val Pro Val Leu Val 1 5 10 15 Glu Leu Asp Gly Asp Val Asn Gly
Gln Lys Phe Ser Val Ser Gly Glu 20 25 30 Gly Glu Gly Asp Ala Thr
Tyr Gly Lys Leu Thr Leu Asn Phe Ile Cys 35 40 45 Thr Thr Gly Lys
Leu Pro Val Pro Trp Pro Thr Leu Val Thr Thr Phe 50 55 60 Ser Tyr
Gly Val Gln Cys Phe Ser Arg Tyr Pro Asp His Met Lys Gln 65 70 75 80
His Asp Phe Phe Lys Ser Ala Met Pro Glu Gly Tyr Val Gln Glu Arg 85
90 95 Thr Ile Phe Tyr Lys Asp Asp Gly Asn Tyr Lys Thr Arg Ala Glu
Val 100 105 110 Lys Phe Glu Gly Asp Thr Leu Val Asn Arg Ile Glu Leu
Lys Gly Ile 115 120 125 Asp Phe Lys Glu Asp Gly Asn Ile Leu Gly His
Lys Met Glu Tyr Asn 130 135 140 Tyr Asn Ser His Asn Val Tyr Ile Met
Gly Asp Lys Pro Lys Asn Gly 145 150 155 160 Ile Lys Val Asn Phe Lys
Ile Arg His Asn Ile Lys Asp Gly Ser Val 165 170 175 Gln Leu Ala Asp
His Tyr Gln Gln Asn Thr Pro Ile Gly Asp Gly Pro 180 185 190 Val Leu
Leu Pro Asp Asn His Tyr Leu Ser Thr Gln Ser Ala Leu Ser 195 200 205
Lys Asp Pro Asn Glu Lys Arg Asp His Met Ile Leu Leu Glu Phe Val 210
215 220 Thr Ala Ala Arg Ile Thr His Gly Met Asp Glu Leu Tyr Lys 225
230 235 57717DNAArtificial SequenceSynthetic DNA sequence encoding
Aequorea victoria GFP1 protein using codons optimized for maize and
Table 1 & Table 2 sequences are maintained 57atg agt aaa ggg
gaa gaa ctt ttc acc ggc gtg gtc cca gtc ctc gtt 48Met Ser Lys Gly
Glu Glu Leu Phe Thr Gly Val Val Pro Val Leu Val 1 5 10 15 gag ttg
gat ggc gat gtg aat ggg caa aaa ttc tct gtc tcc ggg gag 96Glu Leu
Asp Gly Asp Val Asn Gly Gln Lys Phe Ser Val Ser Gly Glu 20 25 30
ggt gag ggt gat gca acc tac gga aag ctg acc cta aat ttt att tgc
144Gly Glu Gly Asp Ala Thr Tyr Gly Lys Leu Thr Leu Asn Phe Ile Cys
35 40 45 acg act ggg aag ttg cct gtg cct tgg ccg aca ctg gtg acg
acg ttc 192Thr Thr Gly Lys Leu Pro Val Pro Trp Pro Thr Leu Val Thr
Thr Phe 50 55 60 tct tat ggt gtg cag tgt ttc tca cgc tac ccg gat
cat atg aaa cag 240Ser Tyr Gly Val Gln Cys Phe Ser Arg Tyr Pro Asp
His Met Lys Gln 65 70 75 80 cat gac ttt ttc aag tcg gcc atg cca gaa
ggc tat gta caa gag aga 288His Asp Phe Phe Lys Ser Ala Met Pro Glu
Gly Tyr Val Gln Glu Arg 85 90 95 act ata ttt tac aag gac gac ggg
aac tac aag aca cgt gct gag gtg 336Thr Ile Phe Tyr Lys Asp Asp Gly
Asn Tyr Lys Thr Arg Ala Glu Val 100 105 110 aag ttc gag ggt gat acc
ctt gtt aat cgg atc gag cta aag ggc att 384Lys Phe Glu Gly Asp Thr
Leu Val Asn Arg Ile Glu Leu Lys Gly Ile 115 120 125 gac ttt aag gag
gac gga aac att ctg gga cac aaa atg gaa tac aac 432Asp Phe Lys Glu
Asp Gly Asn Ile Leu Gly His Lys Met Glu Tyr Asn 130 135 140 tat aac
tcg cac aac gta tac atc atg gga gac aaa cca aag aat ggc 480Tyr Asn
Ser His Asn Val Tyr Ile Met Gly Asp Lys Pro Lys Asn Gly 145 150 155
160 ata aag gtt aac ttc aag att cga cac aac att aaa gac ggc agc gtt
528Ile Lys Val Asn Phe Lys Ile Arg His Asn Ile Lys Asp Gly Ser Val
165 170 175 cag ttg gcc gac cac tat caa caa aat act cca att ggc gat
ggc cct 576Gln Leu Ala Asp His Tyr Gln Gln Asn Thr Pro Ile Gly Asp
Gly Pro 180 185 190 gtc ctc tta ccc gac aac cat tac ctg tcc acg caa
tca gcg ctc agc 624Val Leu Leu Pro Asp Asn His Tyr Leu Ser Thr Gln
Ser Ala Leu Ser 195 200 205 aag gac ccc aac gag aag agg gat cac atg
atc ctc ctt gag ttt gtc 672Lys Asp Pro Asn Glu Lys Arg Asp His Met
Ile Leu Leu Glu Phe Val 210 215 220 acc gca gct agg ata acc cac ggc
atg gat gaa ctg tac aag taa 717Thr Ala Ala Arg Ile Thr His Gly Met
Asp Glu Leu Tyr Lys 225 230 235 58238PRTArtificial
SequenceSynthetic Construct 58Met Ser Lys Gly Glu Glu Leu Phe Thr
Gly Val Val Pro Val Leu Val 1 5 10 15 Glu Leu Asp Gly Asp Val Asn
Gly Gln Lys Phe Ser Val Ser Gly Glu 20 25 30 Gly Glu Gly Asp Ala
Thr Tyr Gly Lys Leu Thr Leu Asn Phe Ile Cys 35 40 45 Thr Thr Gly
Lys Leu Pro Val Pro Trp Pro Thr Leu Val Thr Thr Phe 50 55 60 Ser
Tyr Gly Val Gln Cys Phe Ser Arg Tyr Pro Asp His Met Lys Gln 65 70
75 80 His Asp Phe Phe Lys Ser Ala Met Pro Glu Gly Tyr Val Gln Glu
Arg 85 90 95 Thr Ile Phe Tyr Lys Asp Asp Gly Asn Tyr Lys Thr Arg
Ala Glu Val 100 105 110 Lys Phe Glu Gly Asp Thr Leu Val Asn Arg Ile
Glu Leu Lys Gly Ile 115 120 125 Asp Phe Lys Glu Asp Gly Asn Ile Leu
Gly His Lys Met Glu Tyr Asn 130 135 140 Tyr Asn Ser His Asn Val Tyr
Ile Met Gly Asp Lys Pro Lys Asn Gly 145 150 155 160 Ile Lys Val Asn
Phe Lys Ile Arg His Asn Ile Lys Asp Gly Ser Val 165 170 175 Gln Leu
Ala Asp His Tyr Gln Gln Asn Thr Pro Ile Gly Asp Gly Pro 180 185 190
Val Leu Leu Pro Asp Asn His Tyr Leu Ser Thr Gln Ser Ala Leu Ser 195
200 205 Lys Asp Pro Asn Glu Lys Arg Asp His Met Ile Leu Leu Glu Phe
Val 210 215 220 Thr Ala Ala Arg Ile Thr His Gly Met Asp Glu Leu Tyr
Lys 225 230 235 59717DNAArtificial SequenceSynthetic DNA sequence
in accordance with the invention encoding Aequorea victoria GFP1
protein using codons optimized for maize and with sequences
identified in Table 2 removed and Table 1 sequences are maintained
59atg agt aaa ggg gaa gaa ctt ttc acc ggc gtg gtc cca gtc ctc gtt
48Met Ser Lys Gly Glu Glu Leu Phe Thr Gly Val Val Pro Val Leu Val 1
5 10 15 gag ttg gat ggc gat gtg aat ggg caa aaa ttc tct gtc tcc ggg
gag 96Glu Leu Asp Gly Asp Val Asn Gly Gln Lys Phe Ser Val Ser Gly
Glu 20 25 30 ggt gag ggt gat gca acc tac gga aag ctg acc cta aat
ttc atc tgc 144Gly Glu Gly Asp Ala Thr Tyr Gly Lys Leu Thr Leu Asn
Phe Ile Cys 35 40 45 acg act ggg aag ttg cct gtg cct tgg ccg aca
ctg gtg acg acg ttc 192Thr Thr Gly Lys Leu Pro Val Pro Trp Pro Thr
Leu Val Thr Thr Phe 50 55 60 tct tat ggt gtg cag tgt ttc tca cgc
tac ccg gat cat atg aaa cag 240Ser Tyr Gly Val Gln Cys Phe Ser Arg
Tyr Pro Asp His Met Lys Gln 65 70 75 80 cat gac ttt ttc aag tcg gcc
atg cca gaa ggc tat gta caa gag aga 288His Asp Phe Phe Lys Ser Ala
Met Pro Glu Gly Tyr Val Gln Glu Arg 85 90 95 act atc ttt tac aag
gac gac ggg aac tac aag aca cgt gct gag gtg 336Thr Ile Phe Tyr Lys
Asp Asp Gly Asn Tyr Lys Thr Arg Ala Glu Val 100 105 110 aag ttc gag
ggt gat acc ctt gtt aat cgg atc gag cta aag ggc att 384Lys Phe Glu
Gly Asp Thr Leu Val Asn Arg Ile Glu Leu Lys Gly Ile 115 120 125 gac
ttt aag gag gac gga aac att ctg gga cac aaa atg gaa tac aac 432Asp
Phe Lys Glu Asp Gly Asn Ile Leu Gly His Lys Met Glu Tyr Asn 130 135
140 tat aac tcg cac aac gta tac atc atg gga gac aaa cca aag aat ggc
480Tyr Asn Ser His Asn Val Tyr Ile Met Gly Asp Lys Pro Lys Asn Gly
145 150 155 160 ata aag gtt aac ttc aag att cga cac aac att aaa gac
ggc agc gtt 528Ile Lys Val Asn Phe Lys Ile Arg His Asn Ile Lys Asp
Gly Ser Val 165 170 175 cag ttg gcc gac cac tat caa caa aat act cca
att ggc gat ggc cct 576Gln Leu Ala Asp His Tyr Gln Gln Asn Thr Pro
Ile Gly Asp Gly Pro 180 185 190 gtc ctc tta ccc gac aac cat tac ctg
tcc acg caa tca gcg ctc agc 624Val Leu Leu Pro Asp Asn His Tyr Leu
Ser Thr Gln Ser Ala Leu Ser 195 200 205 aag gac ccc aac gag aag agg
gat cac atg atc ctc ctt gag ttt gtc 672Lys Asp Pro Asn Glu Lys Arg
Asp His Met Ile Leu Leu Glu Phe Val 210 215 220 acc gca gct agg ata
acc cac ggc atg gat gaa ctg tac aag taa 717Thr Ala Ala Arg Ile Thr
His Gly Met Asp Glu Leu Tyr Lys 225 230 235 60238PRTArtificial
SequenceSynthetic Construct 60Met Ser Lys Gly Glu Glu Leu Phe Thr
Gly Val Val Pro Val Leu Val 1 5 10 15 Glu Leu Asp Gly Asp Val Asn
Gly Gln Lys Phe Ser Val Ser Gly Glu 20 25 30 Gly Glu Gly Asp Ala
Thr Tyr Gly Lys Leu Thr Leu Asn Phe Ile Cys 35 40 45 Thr Thr Gly
Lys Leu Pro Val Pro Trp Pro Thr Leu Val Thr Thr Phe 50 55 60 Ser
Tyr Gly Val Gln Cys Phe Ser Arg Tyr Pro Asp His Met Lys Gln 65 70
75 80 His Asp Phe Phe Lys Ser Ala Met Pro Glu Gly Tyr Val Gln Glu
Arg 85 90 95 Thr Ile Phe Tyr Lys Asp Asp Gly Asn Tyr Lys Thr Arg
Ala Glu Val 100 105 110 Lys Phe Glu Gly Asp Thr Leu Val Asn Arg Ile
Glu Leu Lys Gly Ile 115 120 125 Asp Phe Lys Glu Asp Gly Asn Ile Leu
Gly His Lys Met Glu Tyr Asn 130 135 140 Tyr Asn Ser His Asn Val Tyr
Ile Met Gly Asp Lys Pro Lys Asn Gly 145 150 155 160 Ile Lys Val Asn
Phe Lys Ile Arg His Asn Ile Lys Asp Gly Ser Val 165 170 175 Gln Leu
Ala Asp His Tyr Gln Gln Asn Thr Pro Ile Gly Asp Gly Pro 180 185 190
Val Leu Leu Pro Asp Asn His Tyr Leu Ser Thr Gln Ser Ala Leu Ser 195
200 205 Lys Asp Pro Asn Glu Lys Arg Asp His Met Ile Leu Leu Glu Phe
Val 210 215 220 Thr Ala Ala Arg Ile Thr His Gly Met Asp Glu Leu Tyr
Lys 225 230 235 611350DNALeptosphaeria nodorumCDS(1)..(1350)Native
DNA sequence encoding Leptosphaeria nodorum delta-9 fatty acid
desaturase protein 61atg gcg gcc ttg gac agc att cca gag gat aag
gct acc tcg tcg aaa 48Met Ala Ala Leu Asp Ser Ile Pro Glu Asp Lys
Ala Thr Ser Ser Lys 1 5 10 15 tcg act cat att caa tat caa gaa gta
act ttt cgg aac tgg tat aag 96Ser Thr His Ile Gln Tyr Gln Glu Val
Thr Phe Arg Asn Trp Tyr Lys 20 25 30 aag ata aat tgg ctc aac acg
acg ctg gtg gtg ctc ata ccc gct ctt 144Lys Ile Asn Trp Leu Asn Thr
Thr Leu Val Val Leu Ile Pro Ala Leu 35 40 45 gga ctc tac cta aca
cgc acc acg cca ctt aca cga cct acg ctc atc 192Gly Leu Tyr Leu Thr
Arg Thr Thr Pro Leu Thr Arg Pro Thr Leu Ile 50 55 60 tgg tcc gtc
ctg tac tac ttc tgc aca gct ttc ggc atc aca ggc gga 240Trp Ser Val
Leu Tyr Tyr Phe Cys Thr Ala Phe Gly Ile Thr Gly Gly 65 70 75 80 tat
cat cga cta tgg agt cat cgc agc tac tcc gct cgt cta ccg cta 288Tyr
His Arg Leu Trp Ser His Arg Ser Tyr Ser Ala Arg Leu Pro Leu 85 90
95 cgc tta ttc cta gcc ttc aca ggc gcc gga gcc atc caa ggt agt gct
336Arg Leu Phe Leu Ala Phe Thr Gly Ala Gly Ala Ile Gln Gly Ser Ala
100 105 110 cga tgg tgg agc gca aat cac cgc gcc cac cac cga tgg acc
gac aca 384Arg Trp Trp Ser Ala Asn His Arg Ala His His Arg Trp Thr
Asp Thr 115 120 125 atg aag gac ccc tac tcc gtt atg cgc ggc cta tta
ttc tcg cac atc 432Met Lys Asp Pro Tyr Ser Val Met Arg Gly Leu Leu
Phe Ser His Ile 130 135 140 gga tgg atg gta ttg aac agc gac ccc aaa
gtc aaa ggc cga aca gac 480Gly Trp Met Val Leu Asn Ser Asp Pro Lys
Val Lys Gly Arg Thr Asp 145 150 155
160 gtc agt gat ctc gac agc gac ccc gtc gta gtc tgg cag cac aag cac
528Val Ser Asp Leu Asp Ser Asp Pro Val Val Val Trp Gln His Lys His
165 170 175 tac ggc aag tgc ctg ctg ttc gcc gcg tgg ata ttc ccc atg
atc gta 576Tyr Gly Lys Cys Leu Leu Phe Ala Ala Trp Ile Phe Pro Met
Ile Val 180 185 190 gcc ggc ctc gga tgg gga gat tgg tgg gga ggc ctt
gtc tac gcc ggc 624Ala Gly Leu Gly Trp Gly Asp Trp Trp Gly Gly Leu
Val Tyr Ala Gly 195 200 205 atc att cga gcg tgt ttc gtc cag cag gcg
aca ttt tgc gtg aac tct 672Ile Ile Arg Ala Cys Phe Val Gln Gln Ala
Thr Phe Cys Val Asn Ser 210 215 220 ctc gcg cat tgg atc ggc gag cag
ccg ttc gac gac aga cgc acg cct 720Leu Ala His Trp Ile Gly Glu Gln
Pro Phe Asp Asp Arg Arg Thr Pro 225 230 235 240 cga gac cac gtt ttg
aca gcg ttg gta acg atg gga gaa gga tat cat 768Arg Asp His Val Leu
Thr Ala Leu Val Thr Met Gly Glu Gly Tyr His 245 250 255 aac ttc cac
cac gaa ttc cca agc gat tat cgc aac gcg atc atc tgg 816Asn Phe His
His Glu Phe Pro Ser Asp Tyr Arg Asn Ala Ile Ile Trp 260 265 270 tac
caa tac gac cct acc aaa tgg ctc att tac ctc ttc tcc ctc ggc 864Tyr
Gln Tyr Asp Pro Thr Lys Trp Leu Ile Tyr Leu Phe Ser Leu Gly 275 280
285 ccc ttc ccc ctc gca tac tcg ctc aaa acc ttc cgg tcc aat gag att
912Pro Phe Pro Leu Ala Tyr Ser Leu Lys Thr Phe Arg Ser Asn Glu Ile
290 295 300 gaa aaa ggg cgg ttg caa caa caa caa aaa gcc ctg gac aag
aag cgc 960Glu Lys Gly Arg Leu Gln Gln Gln Gln Lys Ala Leu Asp Lys
Lys Arg 305 310 315 320 tca gga ctt gat tgg ggc cta ccc ctc ttc caa
ctc cct gtc ata tcg 1008Ser Gly Leu Asp Trp Gly Leu Pro Leu Phe Gln
Leu Pro Val Ile Ser 325 330 335 tgg gac gac ttc caa gcg cgt tgc aaa
gag tcc ggc gag atg ctg gtt 1056Trp Asp Asp Phe Gln Ala Arg Cys Lys
Glu Ser Gly Glu Met Leu Val 340 345 350 gct gtc gca ggt gtg att cac
gac gtc agc cag ttt att gaa gat cac 1104Ala Val Ala Gly Val Ile His
Asp Val Ser Gln Phe Ile Glu Asp His 355 360 365 cct gga ggc agg agt
ttg att cgg agt gcg gtg ggc aaa gat ggg aca 1152Pro Gly Gly Arg Ser
Leu Ile Arg Ser Ala Val Gly Lys Asp Gly Thr 370 375 380 ggg atg ttt
aat gga ggc gta tat gag cac agt aat gcg gcg cat aat 1200Gly Met Phe
Asn Gly Gly Val Tyr Glu His Ser Asn Ala Ala His Asn 385 390 395 400
ctg ttg tcg aca atg agg gtg gga gtg ctt aga ggt ggg cag gag gtg
1248Leu Leu Ser Thr Met Arg Val Gly Val Leu Arg Gly Gly Gln Glu Val
405 410 415 gag gtg tgg aag aag cag aga gtg gat gtt tta ggg aag agc
gac att 1296Glu Val Trp Lys Lys Gln Arg Val Asp Val Leu Gly Lys Ser
Asp Ile 420 425 430 ttg aga cag gtt acg cgg gtg gag agg ttg gtt gag
ggg gct gtg gct 1344Leu Arg Gln Val Thr Arg Val Glu Arg Leu Val Glu
Gly Ala Val Ala 435 440 445 gcg tag 1350Ala 62449PRTLeptosphaeria
nodorum 62Met Ala Ala Leu Asp Ser Ile Pro Glu Asp Lys Ala Thr Ser
Ser Lys 1 5 10 15 Ser Thr His Ile Gln Tyr Gln Glu Val Thr Phe Arg
Asn Trp Tyr Lys 20 25 30 Lys Ile Asn Trp Leu Asn Thr Thr Leu Val
Val Leu Ile Pro Ala Leu 35 40 45 Gly Leu Tyr Leu Thr Arg Thr Thr
Pro Leu Thr Arg Pro Thr Leu Ile 50 55 60 Trp Ser Val Leu Tyr Tyr
Phe Cys Thr Ala Phe Gly Ile Thr Gly Gly 65 70 75 80 Tyr His Arg Leu
Trp Ser His Arg Ser Tyr Ser Ala Arg Leu Pro Leu 85 90 95 Arg Leu
Phe Leu Ala Phe Thr Gly Ala Gly Ala Ile Gln Gly Ser Ala 100 105 110
Arg Trp Trp Ser Ala Asn His Arg Ala His His Arg Trp Thr Asp Thr 115
120 125 Met Lys Asp Pro Tyr Ser Val Met Arg Gly Leu Leu Phe Ser His
Ile 130 135 140 Gly Trp Met Val Leu Asn Ser Asp Pro Lys Val Lys Gly
Arg Thr Asp 145 150 155 160 Val Ser Asp Leu Asp Ser Asp Pro Val Val
Val Trp Gln His Lys His 165 170 175 Tyr Gly Lys Cys Leu Leu Phe Ala
Ala Trp Ile Phe Pro Met Ile Val 180 185 190 Ala Gly Leu Gly Trp Gly
Asp Trp Trp Gly Gly Leu Val Tyr Ala Gly 195 200 205 Ile Ile Arg Ala
Cys Phe Val Gln Gln Ala Thr Phe Cys Val Asn Ser 210 215 220 Leu Ala
His Trp Ile Gly Glu Gln Pro Phe Asp Asp Arg Arg Thr Pro 225 230 235
240 Arg Asp His Val Leu Thr Ala Leu Val Thr Met Gly Glu Gly Tyr His
245 250 255 Asn Phe His His Glu Phe Pro Ser Asp Tyr Arg Asn Ala Ile
Ile Trp 260 265 270 Tyr Gln Tyr Asp Pro Thr Lys Trp Leu Ile Tyr Leu
Phe Ser Leu Gly 275 280 285 Pro Phe Pro Leu Ala Tyr Ser Leu Lys Thr
Phe Arg Ser Asn Glu Ile 290 295 300 Glu Lys Gly Arg Leu Gln Gln Gln
Gln Lys Ala Leu Asp Lys Lys Arg 305 310 315 320 Ser Gly Leu Asp Trp
Gly Leu Pro Leu Phe Gln Leu Pro Val Ile Ser 325 330 335 Trp Asp Asp
Phe Gln Ala Arg Cys Lys Glu Ser Gly Glu Met Leu Val 340 345 350 Ala
Val Ala Gly Val Ile His Asp Val Ser Gln Phe Ile Glu Asp His 355 360
365 Pro Gly Gly Arg Ser Leu Ile Arg Ser Ala Val Gly Lys Asp Gly Thr
370 375 380 Gly Met Phe Asn Gly Gly Val Tyr Glu His Ser Asn Ala Ala
His Asn 385 390 395 400 Leu Leu Ser Thr Met Arg Val Gly Val Leu Arg
Gly Gly Gln Glu Val 405 410 415 Glu Val Trp Lys Lys Gln Arg Val Asp
Val Leu Gly Lys Ser Asp Ile 420 425 430 Leu Arg Gln Val Thr Arg Val
Glu Arg Leu Val Glu Gly Ala Val Ala 435 440 445 Ala
631350DNAArtificial SequenceSynthetic DNA sequence encoding
Leptosphaeria nodorum delta-9 fatty acid desaturase protein using
codons optimized for maize and Table 1 & Table 2 sequences are
maintained 63atg gca gcc ctt gac agc atc cca gag gat aag gct acc
tcg tct aaa 48Met Ala Ala Leu Asp Ser Ile Pro Glu Asp Lys Ala Thr
Ser Ser Lys 1 5 10 15 tcg act cat att cag tac caa gaa gtg act ttt
cgg aac tgg tac aaa 96Ser Thr His Ile Gln Tyr Gln Glu Val Thr Phe
Arg Asn Trp Tyr Lys 20 25 30 aag ata aac tgg ctc aac acg acg ctg
gtg gtg ctc ata cca gct ctt 144Lys Ile Asn Trp Leu Asn Thr Thr Leu
Val Val Leu Ile Pro Ala Leu 35 40 45 ggt ctt tac cta aca agg acc
acg cca ctt act agg cca acg ctc atc 192Gly Leu Tyr Leu Thr Arg Thr
Thr Pro Leu Thr Arg Pro Thr Leu Ile 50 55 60 tgg tcc gtc ctg tac
tac ttt tgc acc gct ttc ggc att acc ggc gga 240Trp Ser Val Leu Tyr
Tyr Phe Cys Thr Ala Phe Gly Ile Thr Gly Gly 65 70 75 80 tat cat aga
cta tgg agt cat cgc agc tac tcc gct cgt cta ccg ctt 288Tyr His Arg
Leu Trp Ser His Arg Ser Tyr Ser Ala Arg Leu Pro Leu 85 90 95 cgc
ttg ttc ctg gcc ttc act ggc gcc ggg gcc atc caa ggt tca gct 336Arg
Leu Phe Leu Ala Phe Thr Gly Ala Gly Ala Ile Gln Gly Ser Ala 100 105
110 agg tgg tgg agc gca aat cac cgc gcc cat cat agg tgg acc gac aca
384Arg Trp Trp Ser Ala Asn His Arg Ala His His Arg Trp Thr Asp Thr
115 120 125 atg aag gac ccc tac tcc gtt atg cgc ggt cta tta ttc tcg
cac atc 432Met Lys Asp Pro Tyr Ser Val Met Arg Gly Leu Leu Phe Ser
His Ile 130 135 140 ggt tgg atg gtt cta aac agc gac ccc aaa gtc aaa
ggc cgc act gac 480Gly Trp Met Val Leu Asn Ser Asp Pro Lys Val Lys
Gly Arg Thr Asp 145 150 155 160 gtc tca gac cta gat agc gac ccc gtc
gtt gtc tgg cag cac aag cac 528Val Ser Asp Leu Asp Ser Asp Pro Val
Val Val Trp Gln His Lys His 165 170 175 tac ggc aag tgc ctg cta ttt
gcc gca tgg ata ttc ccg atg atc gta 576Tyr Gly Lys Cys Leu Leu Phe
Ala Ala Trp Ile Phe Pro Met Ile Val 180 185 190 gcc ggc ctc gga tgg
gga gat tgg tgg gga ggc ctt gtc tac gcc ggc 624Ala Gly Leu Gly Trp
Gly Asp Trp Trp Gly Gly Leu Val Tyr Ala Gly 195 200 205 atc att agg
gcg tgt ttc gtc cag caa gca acc ttt tgc gtg aac tct 672Ile Ile Arg
Ala Cys Phe Val Gln Gln Ala Thr Phe Cys Val Asn Ser 210 215 220 ctc
gcg cac tgg atc ggc gag cag ccg ttc gac gac aga cgc acc cct 720Leu
Ala His Trp Ile Gly Glu Gln Pro Phe Asp Asp Arg Arg Thr Pro 225 230
235 240 aga gac cac gtt ttg acc gcg ttg gtc act atg gga gaa ggt tat
cac 768Arg Asp His Val Leu Thr Ala Leu Val Thr Met Gly Glu Gly Tyr
His 245 250 255 aac ttc cac cac gag ttc ccg tct gat tat agg aac gcg
atc atc tgg 816Asn Phe His His Glu Phe Pro Ser Asp Tyr Arg Asn Ala
Ile Ile Trp 260 265 270 tat cag tac gac cct acc aaa tgg ctc ata tac
ctc ttc tcc ctc ggc 864Tyr Gln Tyr Asp Pro Thr Lys Trp Leu Ile Tyr
Leu Phe Ser Leu Gly 275 280 285 ccg ttc cca ctg gca tac tcg ctc aaa
acc ttc cgg tct aac gag atc 912Pro Phe Pro Leu Ala Tyr Ser Leu Lys
Thr Phe Arg Ser Asn Glu Ile 290 295 300 gaa aag ggg cgg ttg caa caa
caa caa aag gcc ctg gat aag aag cgc 960Glu Lys Gly Arg Leu Gln Gln
Gln Gln Lys Ala Leu Asp Lys Lys Arg 305 310 315 320 tct ggc ctt gat
tgg ggc ctg ccc ctc ttc cag ctc cct gtg ata tct 1008Ser Gly Leu Asp
Trp Gly Leu Pro Leu Phe Gln Leu Pro Val Ile Ser 325 330 335 tgg gac
gac ttc caa gcg cgt tgt aag gag tcc ggc gag atg ctg gtt 1056Trp Asp
Asp Phe Gln Ala Arg Cys Lys Glu Ser Gly Glu Met Leu Val 340 345 350
gct gtc gcc ggt gtg att cac gac gtc tca cag ttt att gaa gat cac
1104Ala Val Ala Gly Val Ile His Asp Val Ser Gln Phe Ile Glu Asp His
355 360 365 cct gga ggg agg agt ctg att cgg tct gcg gtg ggc aag gat
ggg act 1152Pro Gly Gly Arg Ser Leu Ile Arg Ser Ala Val Gly Lys Asp
Gly Thr 370 375 380 ggg atg ttt aat gga ggc gtt tat gag cac agt aat
gcg gcg cac aat 1200Gly Met Phe Asn Gly Gly Val Tyr Glu His Ser Asn
Ala Ala His Asn 385 390 395 400 ctg ttg tca aca atg agg gtg ggt gtg
ctt aga ggt ggg caa gag gtg 1248Leu Leu Ser Thr Met Arg Val Gly Val
Leu Arg Gly Gly Gln Glu Val 405 410 415 gag gtg tgg aag aag cag cgt
gtg gat gtt tta ggg aag agc gat atc 1296Glu Val Trp Lys Lys Gln Arg
Val Asp Val Leu Gly Lys Ser Asp Ile 420 425 430 ttg cgt caa gtt acg
cgg gtg gag agg ctg gtt gag ggg gct gtg gct 1344Leu Arg Gln Val Thr
Arg Val Glu Arg Leu Val Glu Gly Ala Val Ala 435 440 445 gcc tag
1350Ala 64449PRTArtificial SequenceSynthetic Construct 64Met Ala
Ala Leu Asp Ser Ile Pro Glu Asp Lys Ala Thr Ser Ser Lys 1 5 10 15
Ser Thr His Ile Gln Tyr Gln Glu Val Thr Phe Arg Asn Trp Tyr Lys 20
25 30 Lys Ile Asn Trp Leu Asn Thr Thr Leu Val Val Leu Ile Pro Ala
Leu 35 40 45 Gly Leu Tyr Leu Thr Arg Thr Thr Pro Leu Thr Arg Pro
Thr Leu Ile 50 55 60 Trp Ser Val Leu Tyr Tyr Phe Cys Thr Ala Phe
Gly Ile Thr Gly Gly 65 70 75 80 Tyr His Arg Leu Trp Ser His Arg Ser
Tyr Ser Ala Arg Leu Pro Leu 85 90 95 Arg Leu Phe Leu Ala Phe Thr
Gly Ala Gly Ala Ile Gln Gly Ser Ala 100 105 110 Arg Trp Trp Ser Ala
Asn His Arg Ala His His Arg Trp Thr Asp Thr 115 120 125 Met Lys Asp
Pro Tyr Ser Val Met Arg Gly Leu Leu Phe Ser His Ile 130 135 140 Gly
Trp Met Val Leu Asn Ser Asp Pro Lys Val Lys Gly Arg Thr Asp 145 150
155 160 Val Ser Asp Leu Asp Ser Asp Pro Val Val Val Trp Gln His Lys
His 165 170 175 Tyr Gly Lys Cys Leu Leu Phe Ala Ala Trp Ile Phe Pro
Met Ile Val 180 185 190 Ala Gly Leu Gly Trp Gly Asp Trp Trp Gly Gly
Leu Val Tyr Ala Gly 195 200 205 Ile Ile Arg Ala Cys Phe Val Gln Gln
Ala Thr Phe Cys Val Asn Ser 210 215 220 Leu Ala His Trp Ile Gly Glu
Gln Pro Phe Asp Asp Arg Arg Thr Pro 225 230 235 240 Arg Asp His Val
Leu Thr Ala Leu Val Thr Met Gly Glu Gly Tyr His 245 250 255 Asn Phe
His His Glu Phe Pro Ser Asp Tyr Arg Asn Ala Ile Ile Trp 260 265 270
Tyr Gln Tyr Asp Pro Thr Lys Trp Leu Ile Tyr Leu Phe Ser Leu Gly 275
280 285 Pro Phe Pro Leu Ala Tyr Ser Leu Lys Thr Phe Arg Ser Asn Glu
Ile 290 295 300 Glu Lys Gly Arg Leu Gln Gln Gln Gln Lys Ala Leu Asp
Lys Lys Arg 305 310 315 320 Ser Gly Leu Asp Trp Gly Leu Pro Leu Phe
Gln Leu Pro Val Ile Ser 325 330 335 Trp Asp Asp Phe Gln Ala Arg Cys
Lys Glu Ser Gly Glu Met Leu Val 340 345 350 Ala Val Ala Gly Val Ile
His Asp Val Ser Gln Phe Ile Glu Asp His 355 360 365 Pro Gly Gly Arg
Ser Leu Ile Arg Ser Ala Val Gly Lys Asp Gly Thr 370 375 380 Gly Met
Phe Asn Gly Gly Val Tyr Glu His Ser Asn Ala Ala His Asn 385 390 395
400 Leu Leu Ser Thr Met Arg Val Gly Val Leu Arg Gly Gly Gln Glu Val
405 410 415 Glu Val Trp Lys Lys Gln Arg Val Asp Val Leu Gly Lys Ser
Asp Ile 420 425 430 Leu Arg Gln Val Thr Arg Val Glu Arg Leu Val Glu
Gly Ala Val Ala 435 440 445 Ala 651350DNAArtificial
SequenceSynthetic DNA sequence in accordance with the invention
encoding Leptosphaeria nodorum delta-9 fatty acid desaturase
protein using codons optimized for maize and with sequences
identified in Table 2 removed and Table 1 sequences are maintained
65atg gca gcc ctt gac agc atc cca gag gat aag gct acc tcg tct aaa
48Met Ala Ala Leu Asp Ser Ile Pro Glu Asp Lys Ala Thr Ser Ser Lys 1
5 10 15 tcg act cat att cag tac caa gaa gtg act ttt
cgg aac tgg tac aaa 96Ser Thr His Ile Gln Tyr Gln Glu Val Thr Phe
Arg Asn Trp Tyr Lys 20 25 30 aag ata aac tgg ctc aac acg acg ctg
gtg gtg ctc ata cca gct ctt 144Lys Ile Asn Trp Leu Asn Thr Thr Leu
Val Val Leu Ile Pro Ala Leu 35 40 45 ggt ctt tac cta aca agg acc
acg cca ctt act agg cca acg ctc atc 192Gly Leu Tyr Leu Thr Arg Thr
Thr Pro Leu Thr Arg Pro Thr Leu Ile 50 55 60 tgg tcc gtc ctg tac
tac ttt tgc acc gct ttc ggc att acc ggc gga 240Trp Ser Val Leu Tyr
Tyr Phe Cys Thr Ala Phe Gly Ile Thr Gly Gly 65 70 75 80 tat cat aga
cta tgg agt cat cgc agc tac tcc gct cgt cta ccg ctt 288Tyr His Arg
Leu Trp Ser His Arg Ser Tyr Ser Ala Arg Leu Pro Leu 85 90 95 cgc
ttg ttc ctg gcc ttc act ggc gcc ggg gcc atc caa ggt tca gct 336Arg
Leu Phe Leu Ala Phe Thr Gly Ala Gly Ala Ile Gln Gly Ser Ala 100 105
110 agg tgg tgg agc gca aat cac cgc gcc cat cat agg tgg acc gac aca
384Arg Trp Trp Ser Ala Asn His Arg Ala His His Arg Trp Thr Asp Thr
115 120 125 atg aag gac ccc tac tcc gtt atg cgc ggt ctg tta ttc tcg
cac atc 432Met Lys Asp Pro Tyr Ser Val Met Arg Gly Leu Leu Phe Ser
His Ile 130 135 140 ggt tgg atg gtt cta aac agc gac ccc aaa gtc aaa
ggc cgc act gac 480Gly Trp Met Val Leu Asn Ser Asp Pro Lys Val Lys
Gly Arg Thr Asp 145 150 155 160 gtc tca gac cta gat agc gac ccc gtc
gtt gtc tgg cag cac aag cac 528Val Ser Asp Leu Asp Ser Asp Pro Val
Val Val Trp Gln His Lys His 165 170 175 tac ggc aag tgc ctg cta ttt
gcc gca tgg ata ttc ccg atg atc gta 576Tyr Gly Lys Cys Leu Leu Phe
Ala Ala Trp Ile Phe Pro Met Ile Val 180 185 190 gcc ggc ctc gga tgg
gga gat tgg tgg gga ggc ctt gtc tac gcc ggc 624Ala Gly Leu Gly Trp
Gly Asp Trp Trp Gly Gly Leu Val Tyr Ala Gly 195 200 205 atc att agg
gcg tgt ttc gtc cag caa gca acc ttt tgc gtg aac tct 672Ile Ile Arg
Ala Cys Phe Val Gln Gln Ala Thr Phe Cys Val Asn Ser 210 215 220 ctc
gcg cac tgg atc ggc gag cag ccg ttc gac gac aga cgc acc cct 720Leu
Ala His Trp Ile Gly Glu Gln Pro Phe Asp Asp Arg Arg Thr Pro 225 230
235 240 aga gac cac gtt ttg acc gcg ttg gtc act atg gga gaa ggt tat
cac 768Arg Asp His Val Leu Thr Ala Leu Val Thr Met Gly Glu Gly Tyr
His 245 250 255 aac ttc cac cac gag ttc ccg tct gat tat agg aac gcg
atc atc tgg 816Asn Phe His His Glu Phe Pro Ser Asp Tyr Arg Asn Ala
Ile Ile Trp 260 265 270 tat cag tac gac cct acc aaa tgg ctc ata tac
ctc ttc tcc ctc ggc 864Tyr Gln Tyr Asp Pro Thr Lys Trp Leu Ile Tyr
Leu Phe Ser Leu Gly 275 280 285 ccg ttc cca ctg gca tac tcg ctc aaa
acc ttc cgg tct aac gag atc 912Pro Phe Pro Leu Ala Tyr Ser Leu Lys
Thr Phe Arg Ser Asn Glu Ile 290 295 300 gaa aag ggg cgg ttg caa caa
caa caa aag gcc ctg gat aag aag cgc 960Glu Lys Gly Arg Leu Gln Gln
Gln Gln Lys Ala Leu Asp Lys Lys Arg 305 310 315 320 tct ggc ctt gat
tgg ggc ctg ccc ctc ttc cag ctc cct gtg ata tct 1008Ser Gly Leu Asp
Trp Gly Leu Pro Leu Phe Gln Leu Pro Val Ile Ser 325 330 335 tgg gac
gac ttc caa gcg cgt tgt aag gag tcc ggc gag atg ctg gtt 1056Trp Asp
Asp Phe Gln Ala Arg Cys Lys Glu Ser Gly Glu Met Leu Val 340 345 350
gct gtc gcc ggt gtg att cac gac gtc tca cag ttc att gaa gat cac
1104Ala Val Ala Gly Val Ile His Asp Val Ser Gln Phe Ile Glu Asp His
355 360 365 cct gga ggg agg agt ctg att cgg tct gcg gtg ggc aag gat
ggg act 1152Pro Gly Gly Arg Ser Leu Ile Arg Ser Ala Val Gly Lys Asp
Gly Thr 370 375 380 ggg atg ttt aat gga ggc gtt tat gag cac agt aat
gcg gcg cac aat 1200Gly Met Phe Asn Gly Gly Val Tyr Glu His Ser Asn
Ala Ala His Asn 385 390 395 400 ctg ttg tca aca atg agg gtg ggt gtg
ctt aga ggt ggg caa gag gtg 1248Leu Leu Ser Thr Met Arg Val Gly Val
Leu Arg Gly Gly Gln Glu Val 405 410 415 gag gtg tgg aag aag cag cgt
gtg gat gta tta ggg aag agc gat atc 1296Glu Val Trp Lys Lys Gln Arg
Val Asp Val Leu Gly Lys Ser Asp Ile 420 425 430 ttg cgt caa gtt acg
cgg gtg gag agg ctg gtt gag ggg gct gtg gct 1344Leu Arg Gln Val Thr
Arg Val Glu Arg Leu Val Glu Gly Ala Val Ala 435 440 445 gcc tag
1350Ala 66449PRTArtificial SequenceSynthetic Construct 66Met Ala
Ala Leu Asp Ser Ile Pro Glu Asp Lys Ala Thr Ser Ser Lys 1 5 10 15
Ser Thr His Ile Gln Tyr Gln Glu Val Thr Phe Arg Asn Trp Tyr Lys 20
25 30 Lys Ile Asn Trp Leu Asn Thr Thr Leu Val Val Leu Ile Pro Ala
Leu 35 40 45 Gly Leu Tyr Leu Thr Arg Thr Thr Pro Leu Thr Arg Pro
Thr Leu Ile 50 55 60 Trp Ser Val Leu Tyr Tyr Phe Cys Thr Ala Phe
Gly Ile Thr Gly Gly 65 70 75 80 Tyr His Arg Leu Trp Ser His Arg Ser
Tyr Ser Ala Arg Leu Pro Leu 85 90 95 Arg Leu Phe Leu Ala Phe Thr
Gly Ala Gly Ala Ile Gln Gly Ser Ala 100 105 110 Arg Trp Trp Ser Ala
Asn His Arg Ala His His Arg Trp Thr Asp Thr 115 120 125 Met Lys Asp
Pro Tyr Ser Val Met Arg Gly Leu Leu Phe Ser His Ile 130 135 140 Gly
Trp Met Val Leu Asn Ser Asp Pro Lys Val Lys Gly Arg Thr Asp 145 150
155 160 Val Ser Asp Leu Asp Ser Asp Pro Val Val Val Trp Gln His Lys
His 165 170 175 Tyr Gly Lys Cys Leu Leu Phe Ala Ala Trp Ile Phe Pro
Met Ile Val 180 185 190 Ala Gly Leu Gly Trp Gly Asp Trp Trp Gly Gly
Leu Val Tyr Ala Gly 195 200 205 Ile Ile Arg Ala Cys Phe Val Gln Gln
Ala Thr Phe Cys Val Asn Ser 210 215 220 Leu Ala His Trp Ile Gly Glu
Gln Pro Phe Asp Asp Arg Arg Thr Pro 225 230 235 240 Arg Asp His Val
Leu Thr Ala Leu Val Thr Met Gly Glu Gly Tyr His 245 250 255 Asn Phe
His His Glu Phe Pro Ser Asp Tyr Arg Asn Ala Ile Ile Trp 260 265 270
Tyr Gln Tyr Asp Pro Thr Lys Trp Leu Ile Tyr Leu Phe Ser Leu Gly 275
280 285 Pro Phe Pro Leu Ala Tyr Ser Leu Lys Thr Phe Arg Ser Asn Glu
Ile 290 295 300 Glu Lys Gly Arg Leu Gln Gln Gln Gln Lys Ala Leu Asp
Lys Lys Arg 305 310 315 320 Ser Gly Leu Asp Trp Gly Leu Pro Leu Phe
Gln Leu Pro Val Ile Ser 325 330 335 Trp Asp Asp Phe Gln Ala Arg Cys
Lys Glu Ser Gly Glu Met Leu Val 340 345 350 Ala Val Ala Gly Val Ile
His Asp Val Ser Gln Phe Ile Glu Asp His 355 360 365 Pro Gly Gly Arg
Ser Leu Ile Arg Ser Ala Val Gly Lys Asp Gly Thr 370 375 380 Gly Met
Phe Asn Gly Gly Val Tyr Glu His Ser Asn Ala Ala His Asn 385 390 395
400 Leu Leu Ser Thr Met Arg Val Gly Val Leu Arg Gly Gly Gln Glu Val
405 410 415 Glu Val Trp Lys Lys Gln Arg Val Asp Val Leu Gly Lys Ser
Asp Ile 420 425 430 Leu Arg Gln Val Thr Arg Val Glu Arg Leu Val Glu
Gly Ala Val Ala 435 440 445 Ala 67660DNAXerophyta
viscosaCDS(1)..(660)Native DNA sequence encoding Xerophyta viscosa
PER1 protein 67atg ccg ggg ctc acc att ggc gac acg atc ccc aac ctg
gag ctt gac 48Met Pro Gly Leu Thr Ile Gly Asp Thr Ile Pro Asn Leu
Glu Leu Asp 1 5 10 15 acc acc cag ggt agg atc aaa atc cac gat tac
gtc ggc aac ggc tac 96Thr Thr Gln Gly Arg Ile Lys Ile His Asp Tyr
Val Gly Asn Gly Tyr 20 25 30 gtc atc ttg ttc tca cac cct gga gac
ttc act cct gtc tgc acc acc 144Val Ile Leu Phe Ser His Pro Gly Asp
Phe Thr Pro Val Cys Thr Thr 35 40 45 gaa ctt gga aag atg gct gct
tac gcc gac gag ttc agc aag cgc ggg 192Glu Leu Gly Lys Met Ala Ala
Tyr Ala Asp Glu Phe Ser Lys Arg Gly 50 55 60 gtt aag ctt ctt ggt
ctt tcc tgc gac gat gta cag agc cac aag gag 240Val Lys Leu Leu Gly
Leu Ser Cys Asp Asp Val Gln Ser His Lys Glu 65 70 75 80 tgg atc aag
gat atc gaa gcc tat acg ccg gga tgt cac gta aaa tat 288Trp Ile Lys
Asp Ile Glu Ala Tyr Thr Pro Gly Cys His Val Lys Tyr 85 90 95 cct
atc gcg gcg gac cca acc cgc gag att atc cag cag cta aac atg 336Pro
Ile Ala Ala Asp Pro Thr Arg Glu Ile Ile Gln Gln Leu Asn Met 100 105
110 gta gac cca gac gag aca gag tcc agc aaa tgc gcc gtg cct tcg cga
384Val Asp Pro Asp Glu Thr Glu Ser Ser Lys Cys Ala Val Pro Ser Arg
115 120 125 gct ctg cac atc att ggg ccc gac aag agg atc aag ctg agt
ttc ctg 432Ala Leu His Ile Ile Gly Pro Asp Lys Arg Ile Lys Leu Ser
Phe Leu 130 135 140 tac ccc gcg tcg acg ggg cga aac atg gat gag gtg
ctg agg gca gtg 480Tyr Pro Ala Ser Thr Gly Arg Asn Met Asp Glu Val
Leu Arg Ala Val 145 150 155 160 gag tcg ctc cag cag gcg gca aag cac
aag gtg gca acg ccg gcg aac 528Glu Ser Leu Gln Gln Ala Ala Lys His
Lys Val Ala Thr Pro Ala Asn 165 170 175 tgg aag cct ggt gaa cct gtt
gtg atc aag cct gat gtg tcc agc gag 576Trp Lys Pro Gly Glu Pro Val
Val Ile Lys Pro Asp Val Ser Ser Glu 180 185 190 gag gcc aag aag ctt
ttc ccg cag ggt tat aaa agt gtt gat ctt cca 624Glu Ala Lys Lys Leu
Phe Pro Gln Gly Tyr Lys Ser Val Asp Leu Pro 195 200 205 tcc aag aag
gat tac ctt cgt ttt acg aac gtc tga 660Ser Lys Lys Asp Tyr Leu Arg
Phe Thr Asn Val 210 215 68219PRTXerophyta viscosa 68Met Pro Gly Leu
Thr Ile Gly Asp Thr Ile Pro Asn Leu Glu Leu Asp 1 5 10 15 Thr Thr
Gln Gly Arg Ile Lys Ile His Asp Tyr Val Gly Asn Gly Tyr 20 25 30
Val Ile Leu Phe Ser His Pro Gly Asp Phe Thr Pro Val Cys Thr Thr 35
40 45 Glu Leu Gly Lys Met Ala Ala Tyr Ala Asp Glu Phe Ser Lys Arg
Gly 50 55 60 Val Lys Leu Leu Gly Leu Ser Cys Asp Asp Val Gln Ser
His Lys Glu 65 70 75 80 Trp Ile Lys Asp Ile Glu Ala Tyr Thr Pro Gly
Cys His Val Lys Tyr 85 90 95 Pro Ile Ala Ala Asp Pro Thr Arg Glu
Ile Ile Gln Gln Leu Asn Met 100 105 110 Val Asp Pro Asp Glu Thr Glu
Ser Ser Lys Cys Ala Val Pro Ser Arg 115 120 125 Ala Leu His Ile Ile
Gly Pro Asp Lys Arg Ile Lys Leu Ser Phe Leu 130 135 140 Tyr Pro Ala
Ser Thr Gly Arg Asn Met Asp Glu Val Leu Arg Ala Val 145 150 155 160
Glu Ser Leu Gln Gln Ala Ala Lys His Lys Val Ala Thr Pro Ala Asn 165
170 175 Trp Lys Pro Gly Glu Pro Val Val Ile Lys Pro Asp Val Ser Ser
Glu 180 185 190 Glu Ala Lys Lys Leu Phe Pro Gln Gly Tyr Lys Ser Val
Asp Leu Pro 195 200 205 Ser Lys Lys Asp Tyr Leu Arg Phe Thr Asn Val
210 215 69660DNAArtificial SequenceSynthetic DNA sequence encoding
Xerophyta viscosa PER1 protein using codons optimized for maize and
Table 1 & Table 2 sequences are maintained 69atg cct gga ttg
act att ggt gac aca att ccc aac ttg gag ctg gat 48Met Pro Gly Leu
Thr Ile Gly Asp Thr Ile Pro Asn Leu Glu Leu Asp 1 5 10 15 acg aca
caa ggt cgc atc aag atc cac gac tat gtc ggg aat gga tac 96Thr Thr
Gln Gly Arg Ile Lys Ile His Asp Tyr Val Gly Asn Gly Tyr 20 25 30
gtg att ctc ttc tca cat cct ggt gat ttc act ccg gtg tgt acc acc
144Val Ile Leu Phe Ser His Pro Gly Asp Phe Thr Pro Val Cys Thr Thr
35 40 45 gaa ttg ggc aag atg gct gct tat gcc gac gag ttc tct aag
cgt ggt 192Glu Leu Gly Lys Met Ala Ala Tyr Ala Asp Glu Phe Ser Lys
Arg Gly 50 55 60 gtg aag ctg ctt ggg ttg tcc tgt gat gat gtc caa
tca cat aag gag 240Val Lys Leu Leu Gly Leu Ser Cys Asp Asp Val Gln
Ser His Lys Glu 65 70 75 80 tgg atc aaa gac ata gag gct tac aca cct
ggc tgt cac gta aaa tat 288Trp Ile Lys Asp Ile Glu Ala Tyr Thr Pro
Gly Cys His Val Lys Tyr 85 90 95 ccg att gct gct gat cca acc aga
gaa atc ata cag cag ctg aac atg 336Pro Ile Ala Ala Asp Pro Thr Arg
Glu Ile Ile Gln Gln Leu Asn Met 100 105 110 gtg gac cct gat gag acg
gaa agc tct aag tgc gct gtg cct tct agg 384Val Asp Pro Asp Glu Thr
Glu Ser Ser Lys Cys Ala Val Pro Ser Arg 115 120 125 gca ctt cac atc
ata gga cca gat aag agg atc aag ctg tcc ttc ctc 432Ala Leu His Ile
Ile Gly Pro Asp Lys Arg Ile Lys Leu Ser Phe Leu 130 135 140 tac cct
gcc tct act ggt cgc aac atg gac gaa gtt ctt aga gcc gtt 480Tyr Pro
Ala Ser Thr Gly Arg Asn Met Asp Glu Val Leu Arg Ala Val 145 150 155
160 gag tct ctt cag caa gca gct aaa cac aaa gtt gca act cct gct aac
528Glu Ser Leu Gln Gln Ala Ala Lys His Lys Val Ala Thr Pro Ala Asn
165 170 175 tgg aaa cct ggc gaa cca gtc gtc atc aaa cca gac gtc agc
tcc gag 576Trp Lys Pro Gly Glu Pro Val Val Ile Lys Pro Asp Val Ser
Ser Glu 180 185 190 gag gcc aag aag ctc ttt cct caa ggt tat aaa agc
gtt gat ttg cct 624Glu Ala Lys Lys Leu Phe Pro Gln Gly Tyr Lys Ser
Val Asp Leu Pro 195 200 205 tca aag aag gac tac ttg agg ttc acc aat
gtt tga 660Ser Lys Lys Asp Tyr Leu Arg Phe Thr Asn Val 210 215
70219PRTArtificial SequenceSynthetic Construct 70Met Pro Gly Leu
Thr Ile Gly Asp Thr Ile Pro Asn Leu Glu Leu Asp 1 5 10 15 Thr Thr
Gln Gly Arg Ile Lys Ile His Asp Tyr Val Gly Asn Gly Tyr 20 25 30
Val Ile Leu Phe Ser His Pro Gly Asp Phe Thr Pro Val Cys Thr Thr 35
40 45 Glu Leu Gly Lys Met Ala Ala Tyr Ala Asp Glu Phe Ser Lys Arg
Gly 50 55 60 Val Lys Leu Leu Gly Leu
Ser Cys Asp Asp Val Gln Ser His Lys Glu 65 70 75 80 Trp Ile Lys Asp
Ile Glu Ala Tyr Thr Pro Gly Cys His Val Lys Tyr 85 90 95 Pro Ile
Ala Ala Asp Pro Thr Arg Glu Ile Ile Gln Gln Leu Asn Met 100 105 110
Val Asp Pro Asp Glu Thr Glu Ser Ser Lys Cys Ala Val Pro Ser Arg 115
120 125 Ala Leu His Ile Ile Gly Pro Asp Lys Arg Ile Lys Leu Ser Phe
Leu 130 135 140 Tyr Pro Ala Ser Thr Gly Arg Asn Met Asp Glu Val Leu
Arg Ala Val 145 150 155 160 Glu Ser Leu Gln Gln Ala Ala Lys His Lys
Val Ala Thr Pro Ala Asn 165 170 175 Trp Lys Pro Gly Glu Pro Val Val
Ile Lys Pro Asp Val Ser Ser Glu 180 185 190 Glu Ala Lys Lys Leu Phe
Pro Gln Gly Tyr Lys Ser Val Asp Leu Pro 195 200 205 Ser Lys Lys Asp
Tyr Leu Arg Phe Thr Asn Val 210 215 71660DNAArtificial
SequenceSynthetic DNA sequence in accordance with the invention
encoding Xerophyta viscosa PER1 protein using codons optimized for
maize and with sequences identified in Table 2 removed and Table 1
sequences are maintained 71atg cct gga ttg act att ggt gac aca att
ccc aac ttg gag ctg gat 48Met Pro Gly Leu Thr Ile Gly Asp Thr Ile
Pro Asn Leu Glu Leu Asp 1 5 10 15 acg aca caa ggt cgc atc aag atc
cac gac tat gtc ggg aat gga tac 96Thr Thr Gln Gly Arg Ile Lys Ile
His Asp Tyr Val Gly Asn Gly Tyr 20 25 30 gtg att ctc ttc tca cat
cct ggt gat ttc act ccg gtg tgt acc acc 144Val Ile Leu Phe Ser His
Pro Gly Asp Phe Thr Pro Val Cys Thr Thr 35 40 45 gaa ttg ggc aag
atg gct gct tat gcc gac gag ttc tct aag cgt ggt 192Glu Leu Gly Lys
Met Ala Ala Tyr Ala Asp Glu Phe Ser Lys Arg Gly 50 55 60 gtg aag
ctg ctt ggg ttg tcc tgt gat gat gtc caa tca cat aag gag 240Val Lys
Leu Leu Gly Leu Ser Cys Asp Asp Val Gln Ser His Lys Glu 65 70 75 80
tgg atc aaa gac ata gag gct tac aca cct ggc tgt cac gta aaa tat
288Trp Ile Lys Asp Ile Glu Ala Tyr Thr Pro Gly Cys His Val Lys Tyr
85 90 95 ccg att gct gct gat cca acc aga gaa atc ata cag cag ctg
aac atg 336Pro Ile Ala Ala Asp Pro Thr Arg Glu Ile Ile Gln Gln Leu
Asn Met 100 105 110 gtg gac cct gat gag acg gaa agc tct aag tgc gct
gtg cct tct agg 384Val Asp Pro Asp Glu Thr Glu Ser Ser Lys Cys Ala
Val Pro Ser Arg 115 120 125 gca ctt cac atc ata gga cca gat aag agg
atc aag ctg tcc ttc ctc 432Ala Leu His Ile Ile Gly Pro Asp Lys Arg
Ile Lys Leu Ser Phe Leu 130 135 140 tac cct gcc tct act ggt cgc aac
atg gac gaa gtt ctt aga gcc gtt 480Tyr Pro Ala Ser Thr Gly Arg Asn
Met Asp Glu Val Leu Arg Ala Val 145 150 155 160 gag tct ctt cag caa
gca gct aaa cac aaa gtt gca act cct gct aac 528Glu Ser Leu Gln Gln
Ala Ala Lys His Lys Val Ala Thr Pro Ala Asn 165 170 175 tgg aaa cct
ggc gaa cca gtc gtc atc aaa cca gac gtc agc tcc gag 576Trp Lys Pro
Gly Glu Pro Val Val Ile Lys Pro Asp Val Ser Ser Glu 180 185 190 gag
gcc aag aag ctc ttt cct caa ggt tat aaa agc gtt gat ttg cct 624Glu
Ala Lys Lys Leu Phe Pro Gln Gly Tyr Lys Ser Val Asp Leu Pro 195 200
205 tca aag aag gac tac ttg agg ttc acc aat gtt tga 660Ser Lys Lys
Asp Tyr Leu Arg Phe Thr Asn Val 210 215 72219PRTArtificial
SequenceSynthetic Construct 72Met Pro Gly Leu Thr Ile Gly Asp Thr
Ile Pro Asn Leu Glu Leu Asp 1 5 10 15 Thr Thr Gln Gly Arg Ile Lys
Ile His Asp Tyr Val Gly Asn Gly Tyr 20 25 30 Val Ile Leu Phe Ser
His Pro Gly Asp Phe Thr Pro Val Cys Thr Thr 35 40 45 Glu Leu Gly
Lys Met Ala Ala Tyr Ala Asp Glu Phe Ser Lys Arg Gly 50 55 60 Val
Lys Leu Leu Gly Leu Ser Cys Asp Asp Val Gln Ser His Lys Glu 65 70
75 80 Trp Ile Lys Asp Ile Glu Ala Tyr Thr Pro Gly Cys His Val Lys
Tyr 85 90 95 Pro Ile Ala Ala Asp Pro Thr Arg Glu Ile Ile Gln Gln
Leu Asn Met 100 105 110 Val Asp Pro Asp Glu Thr Glu Ser Ser Lys Cys
Ala Val Pro Ser Arg 115 120 125 Ala Leu His Ile Ile Gly Pro Asp Lys
Arg Ile Lys Leu Ser Phe Leu 130 135 140 Tyr Pro Ala Ser Thr Gly Arg
Asn Met Asp Glu Val Leu Arg Ala Val 145 150 155 160 Glu Ser Leu Gln
Gln Ala Ala Lys His Lys Val Ala Thr Pro Ala Asn 165 170 175 Trp Lys
Pro Gly Glu Pro Val Val Ile Lys Pro Asp Val Ser Ser Glu 180 185 190
Glu Ala Lys Lys Leu Phe Pro Gln Gly Tyr Lys Ser Val Asp Leu Pro 195
200 205 Ser Lys Lys Asp Tyr Leu Arg Phe Thr Asn Val 210 215
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