Rorug04G0222600

50S ribosomal protein L18

Design Primers CRISPR Design

Basic Information

Type: gene
Biological Identity
rosa_rugosa
GWHBQTZ00000004
OG0009178
Physical Location & Seq
Reverse (-)
38224060 .. 38226264
2205 bp
Loading structure...
UTR
Exon/CDS
Intron
Rorug04G0222600.1

Sequence Viewer

Length: 1398 bp
ATGGTGGAGGAGGGAGCGGCGGCTACTCCTTCATTGTTGGCTAAGGCCTTGACGAGCCTGATGGGGGGCGGGGAGATTATCGAAGTCCCTTTCTTCTATCTGTATGGCCGGCTGTTGCTGGAGAGGCCTGCAAGGCTTCTGGTGGTATCGGCGGCGATCTCCGGTGTGAGGAGTTTGAAGGATCTGATCCGTGAAGGGGAGGGGACGACGATCTTTGCTCTTCATGGAGGATTGTGGGTTGATAAGAATCTATTGGCGTCGGAGTTAGAGCCGGGCATTGCTGGGATCGGACTGGTATTGGTGGCGGCCTTGCCTCTGTGGGCGGCCTTCGTGGGCTCGAGGTTGAGTTCATCGGCGTTGTCAAGGTTGGCGGCGGCGGCTGGAGATGGTGCGTTGATGGCGGCTGAGGGTGGGTCCTGGTTCATGTTGGGTTCGCCGGCGGGGCTGCTGGATGCTGGGCTTAGCAGGGCGATGGTTGGAGCGGCGGCGAGGATTGCAGCAGGTGCTGTGCTGCTGTCTCAGTGTATCTCGGCGGGAAACTCTCTTCTGAGAAACCCTAATGGGTCAGAGGATAGTGGGCCTGGGTCTGGGATGGAGTTGGGTCAAGGGTTTGTGGAGATTGGGCCGGATGCAGCTGATGCTGGGGTTGGATTAACAGGGGTGCTGAAAATTGCAGCGGCTAAATATACCCACATAAACCAAATAAAGGAGGAAGTTTATGGTACGTTGGTGGCAGATAACTCCATTGCAGTAAACCATGATCACTTCTTGACCTACCATATTGACCTTGATGTGGATGGTGAAGCCAACTCCTTTGTGAAGAACAATTTCGTGACAAAACGAGTTACAGACCCAAACATACCGAGGAAGAGTTACTGGAATGTTGTAAGCGAAACCGCTAAGACGGAAGCGGATGCACGGATTCACCTAGGCTTGAAGCCAGCCGACCTGGTTGTGGTAAATCCTAATAAGAAGACCAAACCCGGGAACCCTATTGGTTACCGTTTGATTCCAGGGTCATTTGTAAATCCCCTTCTGGTGGAGGATGACTACCCACAAATTCGTGGTGCTTTCACCAACTATAATGTTTGGGTTACACCGTACAACAAGTCTGAAAAATGGGCGGGAGGACCATATGTTGATCAAAGCCGAGGAGATGATACCTTAGCAGTCTGGAGCCAAAGGAATAGGAAGATTGAGAACAAGGACATAGTGTTGTGGTACACCGTTGGGTTTCACCACAATCCTTGCCAAGAGGACTTTCCGATGATGCCAACATTGAGCGGTGGATTTGAGCTCCGGCCAGCTAATTTCTTTGAGAGCAGTCCAGTTCTTAAAGTTAGAACAACACCTCCTCCTAAGCATGTACCATCTCCTAATTGCACCAAGCAAACCTAG

Protein Analysis

465

Amino Acids

49.6

Weight (kDa)

6.03

Isoelectric Point (pI)

33.97

Instability Index
Protein Domains (Pfam)
Domain Name Pfam ID Position E-value Description
Cu_amine_oxid PF01179 215 - 445 4.1e-69 Copper amine oxidase, enzyme domain
Hydrophobicity Profile (Kyte-Doolittle)
AI Structure Prediction Report
Calculating structure properties...

Genomic Context

Gene Family Tree

Species Filter
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Publication-ready
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Restriction Enzyme Sites

1 / 10
Enzyme Recognition Site Cut Count Positions (bp)
AarI CACCTGC 1 cut(s) 491
Acc36I ACCTGC 1 cut(s) 491
AccBSI CCGCTC 3 cut(s) 17, 482, 1284
AclWI GGATC 3 cut(s) 181, 189, 293
AcoI YGGCCR 2 cut(s) 106, 1301
AcsI RAATTY 1 cut(s) 1059
AcyI GRCGYC 1 cut(s) 257
AfaI GTAC 4 cut(s) 724, 1103, 1223, 1368
AfiI CCNNNNNNNGG 9 cut(s) 64, 168, 196, 587, 706, 793, 955, 984, 1039
AgsI TTSAA 2 cut(s) 178, 937
AjnI CCWGG 4 cut(s) 416, 580, 948, 1012
AjuI GAANNNNNNNTTGG 2 cut(s) 1245, 1277
AloI GAACNNNNNNTCC 2 cut(s) 1336, 1368
AluBI AGCT 3 cut(s) 635, 1297, 1307
AluI AGCT 3 cut(s) 635, 1297, 1307
Alw21I GWGCWC 1 cut(s) 1299
Alw26I GTCTC 1 cut(s) 522
AlwI GGATC 3 cut(s) 181, 189, 293
AlwNI CAGNNNCTG 1 cut(s) 506
Ama87I CYCGRG 2 cut(s) 337, 983
AoxI GGCC 8 cut(s) 45, 106, 125, 306, 324, 578, 623, 1301
ApeKI GCWGC 5 cut(s) 445, 497, 511, 632, 674
ApoI RAATTY 1 cut(s) 1059
Asp700I GAANNNNTTC 1 cut(s) 827
AspA2I CCTAGG 1 cut(s) 928
AspS9I GGNCC 4 cut(s) 414, 578, 623, 1130
AsuC2I CCSGG 3 cut(s) 273, 984, 985
AsuHPI GGTGA 4 cut(s) 812, 917, 1066, 1229
AvaI CYCGRG 2 cut(s) 337, 983
AvaII GGWCC 2 cut(s) 414, 1130
AvrII CCTAGG 1 cut(s) 928
BaeI ACNNNNGTAYC 2 cut(s) 714, 747
BanII GRGCYC 2 cut(s) 338, 1299
BarI GAAGNNNNNNTAC 2 cut(s) 1017, 1049
BbsI GAAGAC 1 cut(s) 980
Bbv12I GWGCWC 1 cut(s) 1299
BbvCI CCTCAGC 1 cut(s) 405
BbvI GCAGC 5 cut(s) 432, 498, 509, 644, 686
BccI CCATC 7 cut(s) 55, 380, 391, 466, 586, 791, 1378
BciT130I CCWGG 4 cut(s) 418, 582, 950, 1014
BclI TGATCA 2 cut(s) 760, 1141
BcnI CCSGG 3 cut(s) 273, 984, 985
BcoDI GTCTC 1 cut(s) 522
BfaI CTAG 2 cut(s) 929, 1396
BfuAI ACCTGC 1 cut(s) 491
BglI GCCNNNNNGGC 2 cut(s) 133, 442
BlnI CCTAGG 1 cut(s) 928
BlpI GCTNAGC 1 cut(s) 461
Bme1390I CCNGG 7 cut(s) 273, 418, 582, 950, 984, 985, 1014
Bme18I GGWCC 2 cut(s) 414, 1130
BmeT110I CYCGRG 2 cut(s) 337, 983
BmgT120I GGNCC 4 cut(s) 414, 578, 623, 1130
BmiI GGNNCC 3 cut(s) 415, 989, 1178
BmrFI CCNGG 7 cut(s) 273, 418, 582, 950, 984, 985, 1014
BmsI GCATC 5 cut(s) 442, 619, 628, 904, 1260
BpiI GAAGAC 1 cut(s) 980
BpmI CTGGAG 3 cut(s) 140, 402, 1195
Bpu10I CCTNAGC 4 cut(s) 42, 405, 1165, 1359
Bpu1102I GCTNAGC 1 cut(s) 461
BpuMI CCSGG 3 cut(s) 273, 984, 985
BsaBI GATNNNNATC 1 cut(s) 246
BsaHI GRCGYC 1 cut(s) 257
BsaJI CCNNGG 6 cut(s) 581, 863, 928, 983, 1013, 1150
BsaWI WCCGGW 1 cut(s) 161
BsaXI ACNNNNNCTCC 4 cut(s) 1336, 1339, 1366, 1369
Bsc4I CCNNNNNNNGG 9 cut(s) 64, 168, 196, 587, 706, 793, 955, 984, 1039
Bse118I RCCGGY 2 cut(s) 108, 436
Bse1I ACTGG 3 cut(s) 297, 881, 1328
Bse3DI GCAATG 2 cut(s) 276, 744
Bse8I GATNNNNATC 1 cut(s) 246
BseBI CCWGG 4 cut(s) 418, 582, 950, 1014
BseDI CCNNGG 6 cut(s) 581, 863, 928, 983, 1013, 1150
BseGI GGATG 6 cut(s) 457, 597, 634, 802, 919, 1051
BseJI GATNNNNATC 1 cut(s) 246
BseLI CCNNNNNNNGG 9 cut(s) 64, 168, 196, 587, 706, 793, 955, 984, 1039
BseMI GCAATG 2 cut(s) 276, 744
BseMII CTCAG 3 cut(s) 396, 533, 539
BseNI ACTGG 3 cut(s) 297, 881, 1328
BseRI GAGGAG 4 cut(s) 23, 184, 1167, 1344
BseXI GCAGC 5 cut(s) 432, 498, 509, 644, 686
BseYI CCCAGC 3 cut(s) 281, 455, 641
BshFI GGCC 8 cut(s) 47, 108, 127, 308, 326, 580, 625, 1303
BsiHKAI GWGCWC 1 cut(s) 1299
BsiHKCI CYCGRG 2 cut(s) 337, 983
BsiSI CCGG 7 cut(s) 109, 162, 272, 437, 626, 984, 1300
BslFI GGGAC 2 cut(s) 71, 217
BslI CCNNNNNNNGG 9 cut(s) 64, 168, 196, 587, 706, 793, 955, 984, 1039
BsmAI GTCTC 1 cut(s) 522
BsmFI GGGAC 2 cut(s) 71, 217
BsnI GGCC 8 cut(s) 47, 108, 127, 308, 326, 580, 625, 1303
BsoBI CYCGRG 2 cut(s) 337, 983
Bsp1286I GDGCHC 2 cut(s) 338, 1299
Bsp143I GATC 7 cut(s) 156, 181, 186, 210, 285, 760, 1141
Bsp1720I GCTNAGC 1 cut(s) 461
BspANI GGCC 8 cut(s) 47, 108, 127, 308, 326, 580, 625, 1303
BspCNI CTCAG 3 cut(s) 397, 532, 540
BspLI GGNNCC 3 cut(s) 415, 989, 1178
BspMI ACCTGC 1 cut(s) 491
BspPI GGATC 3 cut(s) 181, 189, 293
BspQI GCTCTTC 1 cut(s) 225
BsrBI CCGCTC 3 cut(s) 17, 482, 1284
BsrDI GCAATG 2 cut(s) 276, 744
BsrFI RCCGGY 2 cut(s) 108, 436
BsrI ACTGG 3 cut(s) 297, 881, 1328
BssAI RCCGGY 2 cut(s) 108, 436
BssECI CCNNGG 6 cut(s) 581, 863, 928, 983, 1013, 1150
BssMI GATC 7 cut(s) 156, 181, 186, 210, 285, 760, 1141
BssNI GRCGYC 1 cut(s) 257
BssT1I CCWWGG 1 cut(s) 928
Bst2UI CCWGG 4 cut(s) 418, 582, 950, 1014
Bst4CI ACNGT 3 cut(s) 1004, 1101, 1228
Bst6I CTCTTC 3 cut(s) 225, 549, 863
BstACI GRCGYC 1 cut(s) 257
BstAPI GCANNNNNTGC 2 cut(s) 503, 638
BstC8I GCNNGC 5 cut(s) 110, 129, 438, 942, 1305
BstDEI CTNAG 8 cut(s) 42, 405, 461, 519, 548, 900, 1165, 1359
BstEII GGTNACC 1 cut(s) 998
BstF5I GGATG 6 cut(s) 457, 597, 634, 802, 919, 1051
BstKTI GATC 7 cut(s) 159, 184, 189, 213, 288, 763, 1144
BstMAI GTCTC 1 cut(s) 522
BstMBI GATC 7 cut(s) 156, 181, 186, 210, 285, 760, 1141
BstMWI GCNNNNNNNGC 8 cut(s) 124, 133, 377, 398, 442, 494, 503, 638
BstNI CCWGG 4 cut(s) 418, 582, 950, 1014
BstNSI RCATGY 1 cut(s) 1367
BstPI GGTNACC 1 cut(s) 998
BstSCI CCNGG 7 cut(s) 271, 416, 580, 948, 982, 983, 1012
BstV1I GCAGC 5 cut(s) 432, 498, 509, 644, 686
BstV2I GAAGAC 1 cut(s) 980
BstX2I RGATCY 1 cut(s) 181
BstYI RGATCY 1 cut(s) 181
BsuRI GGCC 8 cut(s) 47, 108, 127, 308, 326, 580, 625, 1303
BtgZI GCGATG 1 cut(s) 485
BtsCI GGATG 6 cut(s) 457, 597, 634, 802, 919, 1051
BtsIMutI CAGTG 1 cut(s) 527
BveI ACCTGC 1 cut(s) 491
Cac8I GCNNGC 5 cut(s) 110, 129, 438, 942, 1305
CaiI CAGNNNCTG 1 cut(s) 506
Cfr10I RCCGGY 2 cut(s) 108, 436
Cfr13I GGNCC 4 cut(s) 414, 578, 623, 1130
Cfr9I CCCGGG 1 cut(s) 983
CseI GACGC 1 cut(s) 246
CsiI ACCWGGT 1 cut(s) 948
Csp6I GTAC 4 cut(s) 723, 1102, 1222, 1367
CspCI CAANNNNNGTGG 2 cut(s) 1229, 1264
CviAII CATG 4 cut(s) 224, 424, 758, 1364
CviQI GTAC 4 cut(s) 723, 1102, 1222, 1367
DdeI CTNAG 8 cut(s) 42, 405, 461, 519, 548, 900, 1165, 1359
DpnI GATC 7 cut(s) 158, 183, 188, 212, 287, 762, 1143
DpnII GATC 7 cut(s) 156, 181, 186, 210, 285, 760, 1141
EaeI YGGCCR 2 cut(s) 106, 1301
Eam1104I CTCTTC 3 cut(s) 225, 549, 863
EarI CTCTTC 3 cut(s) 225, 549, 863
Ecl136II GAGCTC 1 cut(s) 1297
Eco130I CCWWGG 1 cut(s) 928
Eco147I AGGCCT 2 cut(s) 47, 127
Eco24I GRGCYC 2 cut(s) 338, 1299
Eco47I GGWCC 2 cut(s) 414, 1130
Eco53kI GAGCTC 1 cut(s) 1297
Eco88I CYCGRG 2 cut(s) 337, 983
Eco91I GGTNACC 1 cut(s) 998
EcoICRI GAGCTC 1 cut(s) 1297
EcoO109I RGGNCCY 1 cut(s) 414
EcoO65I GGTNACC 1 cut(s) 998
EcoRII CCWGG 4 cut(s) 416, 580, 948, 1012
EcoT14I CCWWGG 1 cut(s) 928
EcoT38I GRGCYC 2 cut(s) 338, 1299
ErhI CCWWGG 1 cut(s) 928
FaeI CATG 4 cut(s) 227, 427, 761, 1367
FaqI GGGAC 2 cut(s) 71, 217
FatI CATG 4 cut(s) 223, 423, 757, 1363
FauI CCCGC 4 cut(s) 62, 433, 526, 1117
FauNDI CATATG 1 cut(s) 1135
FbaI TGATCA 2 cut(s) 760, 1141
FokI GGATG 6 cut(s) 464, 604, 641, 809, 926, 1058
FriOI GRGCYC 2 cut(s) 338, 1299
FspBI CTAG 2 cut(s) 929, 1396
GsaI CCCAGC 3 cut(s) 285, 459, 645
GsuI CTGGAG 3 cut(s) 140, 402, 1195
HaeIII GGCC 8 cut(s) 47, 108, 127, 308, 326, 580, 625, 1303
HapII CCGG 7 cut(s) 109, 162, 272, 437, 626, 984, 1300
HgaI GACGC 1 cut(s) 246
Hin1I GRCGYC 1 cut(s) 257
Hin1II CATG 4 cut(s) 227, 427, 761, 1367
HinfI GANTC 3 cut(s) 247, 922, 1009
HpaII CCGG 7 cut(s) 109, 162, 272, 437, 626, 984, 1300
HphI GGTGA 4 cut(s) 812, 917, 1066, 1229
Hpy166II GTNNAC 2 cut(s) 754, 1224
Hpy188I TCNGA 7 cut(s) 186, 262, 290, 549, 568, 1114, 1266
Hpy188III TCNNGA 3 cut(s) 769, 832, 1174
Hpy8I GTNNAC 2 cut(s) 754, 1224
Hpy99I CGWCG 2 cut(s) 211, 262
HpyAV CCTTC 5 cut(s) 39, 172, 188, 337, 1043
HpyCH4III ACNGT 3 cut(s) 1004, 1101, 1228
HpyCH4IV ACGT 1 cut(s) 725
HpyCH4V TGCA 7 cut(s) 131, 497, 632, 674, 749, 917, 1383
HpyF10VI GCNNNNNNNGC 8 cut(s) 124, 133, 377, 398, 442, 494, 503, 638
HpyF3I CTNAG 8 cut(s) 42, 405, 461, 519, 548, 900, 1165, 1359
HpySE526I ACGT 1 cut(s) 725
Hsp92I GRCGYC 1 cut(s) 257
Hsp92II CATG 4 cut(s) 227, 427, 761, 1367
KroI GCCGGC 2 cut(s) 108, 436
KroNI GCCGGC 2 cut(s) 110, 438
Ksp22I TGATCA 2 cut(s) 760, 1141
Kzo9I GATC 7 cut(s) 156, 181, 186, 210, 285, 760, 1141
LguI GCTCTTC 1 cut(s) 225
LmnI GCTCC 4 cut(s) 14, 479, 1176, 1302
Lsp1109I GCAGC 5 cut(s) 432, 498, 509, 644, 686
LweI GCATC 5 cut(s) 442, 619, 628, 904, 1260
MabI ACCWGGT 1 cut(s) 948
MaeI CTAG 2 cut(s) 929, 1396
MaeII ACGT 1 cut(s) 725
MaeIII GTNAC 5 cut(s) 832, 844, 872, 998, 1093
MalI GATC 7 cut(s) 158, 183, 188, 212, 287, 762, 1143
MbiI CCGCTC 3 cut(s) 17, 482, 1284
MboI GATC 7 cut(s) 156, 181, 186, 210, 285, 760, 1141
MboII GAAGA 7 cut(s) 85, 212, 536, 832, 880, 985, 1204
MflI RGATCY 1 cut(s) 181
MhlI GDGCHC 2 cut(s) 338, 1299
MluCI AATT 5 cut(s) 669, 826, 1059, 1309, 1378
MmeI TCCRAC 3 cut(s) 240, 457, 628
MreI CGCCGGCG 1 cut(s) 436
MroNI GCCGGC 2 cut(s) 108, 436
MroXI GAANNNNTTC 1 cut(s) 827
MseI TTAA 2 cut(s) 653, 1335
MspA1I CMGCKG 2 cut(s) 635, 677
MspI CCGG 7 cut(s) 109, 162, 272, 437, 626, 984, 1300
MspR9I CCNGG 7 cut(s) 273, 418, 582, 950, 984, 985, 1014
MvaI CCWGG 4 cut(s) 418, 582, 950, 1014
MwoI GCNNNNNNNGC 8 cut(s) 124, 133, 377, 398, 442, 494, 503, 638
NaeI GCCGGC 2 cut(s) 110, 438
NciI CCSGG 3 cut(s) 273, 984, 985
NdeI CATATG 1 cut(s) 1135
NdeII GATC 7 cut(s) 156, 181, 186, 210, 285, 760, 1141
NgoMIV GCCGGC 2 cut(s) 108, 436
NlaIII CATG 4 cut(s) 227, 427, 761, 1367
NlaIV GGNNCC 3 cut(s) 415, 989, 1178
NmeAIII GCCGAG 2 cut(s) 509, 1175
NmuCI GTSAC 1 cut(s) 832
NspI RCATGY 1 cut(s) 1367
PaeR7I CTCGAG 1 cut(s) 337
PaqCI CACCTGC 1 cut(s) 491
PceI AGGCCT 2 cut(s) 47, 127
PciSI GCTCTTC 1 cut(s) 225
PdiI GCCGGC 2 cut(s) 110, 438
PdmI GAANNNNTTC 1 cut(s) 827
PfeI GAWTC 3 cut(s) 247, 922, 1009
PpuMI RGGWCCY 1 cut(s) 414
Psp124BI GAGCTC 1 cut(s) 1299
Psp5II RGGWCCY 1 cut(s) 414
Psp6I CCWGG 4 cut(s) 416, 580, 948, 1012
PspEI GGTNACC 1 cut(s) 998
PspFI CCCAGC 3 cut(s) 281, 455, 641
PspGI CCWGG 4 cut(s) 416, 580, 948, 1012
PspN4I GGNNCC 3 cut(s) 415, 989, 1178
PspPI GGNCC 4 cut(s) 414, 578, 623, 1130
PspPPI RGGWCCY 1 cut(s) 414
PspXI VCTCGAGB 1 cut(s) 337
PstNI CAGNNNCTG 1 cut(s) 506
PsuI RGATCY 1 cut(s) 181
PvuII CAGCTG 1 cut(s) 635
RsaI GTAC 4 cut(s) 724, 1103, 1223, 1368
RsaNI GTAC 4 cut(s) 723, 1102, 1222, 1367
SacI GAGCTC 1 cut(s) 1299
SapI GCTCTTC 1 cut(s) 225
SaqAI TTAA 2 cut(s) 653, 1335
Sau3AI GATC 7 cut(s) 156, 181, 186, 210, 285, 760, 1141
Sau96I GGNCC 4 cut(s) 414, 578, 623, 1130
ScrFI CCNGG 7 cut(s) 273, 418, 582, 950, 984, 985, 1014
SduI GDGCHC 2 cut(s) 338, 1299
SexAI ACCWGGT 1 cut(s) 948
SfaNI GCATC 5 cut(s) 442, 619, 628, 904, 1260
Sfr274I CTCGAG 1 cut(s) 337
SgrAI CRCCGGYG 1 cut(s) 436
SinI GGWCC 2 cut(s) 414, 1130
SlaI CTCGAG 1 cut(s) 337
SmaI CCCGGG 1 cut(s) 985
SmlI CTYRAG 1 cut(s) 337
SmoI CTYRAG 1 cut(s) 337
Sse9I AATT 5 cut(s) 669, 826, 1059, 1309, 1378
SseBI AGGCCT 2 cut(s) 47, 127
SspMI CTAG 2 cut(s) 929, 1396
SstI GAGCTC 1 cut(s) 1299
StuI AGGCCT 2 cut(s) 47, 127
StyD4I CCNGG 7 cut(s) 271, 416, 580, 948, 982, 983, 1012
StyI CCWWGG 1 cut(s) 928
TaaI ACNGT 3 cut(s) 1004, 1101, 1228
TaiI ACGT 1 cut(s) 728
TaqI TCGA 2 cut(s) 81, 338
TasI AATT 5 cut(s) 669, 826, 1059, 1309, 1378
TfiI GAWTC 3 cut(s) 247, 922, 1009
Tru1I TTAA 2 cut(s) 653, 1335
Tru9I TTAA 2 cut(s) 653, 1335
TscAI CASTG 1 cut(s) 527
TseFI GTSAC 1 cut(s) 832
TseI GCWGC 5 cut(s) 445, 497, 511, 632, 674
Tsp45I GTSAC 1 cut(s) 832
TspDTI ATGAA 4 cut(s) 21, 212, 339, 412
TspGWI ACGGA 3 cut(s) 179, 920, 934
TspMI CCCGGG 1 cut(s) 983
TspRI CASTG 1 cut(s) 527
VpaK11BI GGWCC 2 cut(s) 414, 1130
XapI RAATTY 1 cut(s) 1059
XceI RCATGY 1 cut(s) 1367
XhoI CTCGAG 1 cut(s) 337
XmaI CCCGGG 1 cut(s) 983
XmaJI CCTAGG 1 cut(s) 928
XmnI GAANNNNTTC 1 cut(s) 827
XspI CTAG 2 cut(s) 929, 1396
Using CommOnly database (standard laboratory enzymes). Scanned on CDS sequence.