FvH4_4g21940

Protein of unknown function (DUF1350)

Design Primers CRISPR Design

Basic Information

Type: gene
Biological Identity
fragaria_vesca
Fvb4
OG0013149
Physical Location & Seq
Forward (+)
24910483 .. 24914599
4117 bp
Loading structure...
UTR
Exon/CDS
Intron
FvH4_4g21940.t1

Sequence Viewer

Length: 1320 bp
ATGGCTAGCTGGGTTAGGAGTGCACAGCTTCATTCTTATCCTGGTGTTCATCATCATCAATCTTTGAAGAAGAGGATTGAGATTCAGCAGCAGGGTCAGTTTCTGAGGTTACCCAATTCTCATAGGGGTCATCTCCGGGGGAGAGTCTTCTGTAACTTTGATGATGAACTTGCCAAGAAAGAATCGCAGCCACAACCAACTGGGGTTCAGCTCTACTCTGAGATTGAGAGGTTGCTTACTGAGACTGTGAGGCAATCACAGGGTGGTTGGAATGATTCCAGTGACTGGAGTCAAGTAGAGGGGGCATGGGTTCTGAAACCGAAAACCTTGAAACCGACGGCAGTTGTACATTTTATCGGTGGTATATTTGTTGGAGCTGCCCCTCAGCTCACCTACCGGTTCTTTCTTGAGCGTCTGGCCGAGAAGGGAATTTTGGTGATTGCAACACCATATCCTAGTGGGTTTGATCACTTCTTCATTGCAGATCAAGTTCAATTCAAATTTGATCGATGTTATAGGTTTTTTAAAGACACAATTGAAGATCTGCCTATCTTTGGCCTTGGGCATTCTTTGGGATCTGTCATCCACCTCTTGATTGGTTCAAGATATGCTGTTCAAAGAAACGGGAATGTATTGATGGCATTCAACAATAGGGAGGCGAGCTATGCTGTTCCTCTGTTCTCACCTGTTCTTGTTCCAGTCGCCCAACGCCTTGGACCAATTTTATCACAGATTGCATCATCACCAACATTTCGCCTTGGGGCTGAAATGACTATGCGGAATTTTGAAAACCTTAGCCCTCCGATCATGAAGCAAGTTCTGCCTTTAGTGGAGCAGCTTCCACCCTTGTATATGGAGCTAGTCAATGGAAGAGAAGATTTTGTCCCAAAACCTGAAGAGACTCGCCGACTTATAAAATCATACTACGGTATTTCCAGGAATCTTCTAGTGAGGTTCAAGGATGATACAATTGACGAGACTTCAATATTAGCACAGGTGCTGAGTTCAGATTCAGCCATCAGTTCTATGCTAGACATGTCAACTCGCCTATTGCCTGGAGATCATGCGCTTCCTTTGCAACAGGCTATACCAGATGTGCCTCCAGCAATGGCAGATGCAGTAAATCGCGGGAGTGAGATGTTGGCCAATCTTACTGTAGGAACGCCATGGGAGTCTGTTGCTAAAGAAGTAGGCAACTCATTAGGTGTAGATTCAACGAAGATCCTTCGTGCAGAAGTGGCCAAAGACCTAGACCTTCTTGTGGATGTTGTTGCATCTTGGATGGCTTCCAATACAGGTCCAAGGCTTCTGAAGCCATGA
Functional Annotation
Gene Ontology
Molecular Function
Biological Process
Cellular Component
No ontology terms assigned.
KEGG Pathways
Metabolic & Signaling
No pathways identified.
Pfam Domains
Protein Families
DUF1350

Protein Analysis

440

Amino Acids

49.03

Weight (kDa)

7.2

Isoelectric Point (pI)

43.23

Instability Index
Protein Domains (Pfam)
Domain Name Pfam ID Position E-value Description
DUF1350 PF07082 95 - 363 9.2e-55 Protein of unknown function (DUF1350)
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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Orthologous Genes (Group: OG0013149)

Species Orthologous Gene IDs
arabidopsis_thaliana AT5G47860
fragaria_vesca FvH4_4g21940
malus_domestica MD13G1135500.v1.1 MD16G1131100.v1.1
prunus_persica Prupe.1G209800_v2.0.a1
pyrus_communis pycom13g11820 pycom16g11560
rosa_chinensis RchiOBHm_Chr4g0428161
rosa_laevigata RLG00000007174
rosa_multiflora Rmu_sc0004940.1_g000008 Rmu_sc0009599.1_g000008
rosa_roxburghii Rroxscaffold_5G00369890
rosa_rugosa Rorug04G0223100
rosa_samantha Rh4AG280400 Rh4BG285900 Rh4CG301200 Rh4DG282900
rosa_wichuraiana Rw4G024350

Restriction Enzyme Sites

1 / 10
Enzyme Recognition Site Cut Count Positions (bp)
AanI TTATAA 1 cut(s) 914
AccB7I CCANNNNNTGG 1 cut(s) 285
AccII CGCG 1 cut(s) 1128
AciI CCGC 2 cut(s) 778, 1128
AclWI GGATC 2 cut(s) 583, 1216
AcoI YGGCCR 3 cut(s) 417, 1143, 1239
AcsI RAATTY 3 cut(s) 429, 500, 781
AcuI CTGAAG 1 cut(s) 915
AdeI CACNNNGTG 1 cut(s) 263
AfaI GTAC 1 cut(s) 348
AfiI CCNNNNNNNGG 3 cut(s) 285, 554, 1261
AflIII ACRYGT 1 cut(s) 1035
AgeI ACCGGT 1 cut(s) 396
AjnI CCWGG 3 cut(s) 40, 935, 1054
AjuI GAANNNNNNNTTGG 2 cut(s) 416, 448
AluBI AGCT 8 cut(s) 9, 28, 211, 377, 388, 663, 838, 859
AluI AGCT 8 cut(s) 9, 28, 211, 377, 388, 663, 838, 859
Alw21I GWGCWC 1 cut(s) 25
Alw26I GTCTC 3 cut(s) 236, 893, 971
Alw44I GTGCAC 1 cut(s) 21
AlwI GGATC 2 cut(s) 583, 1216
AlwNI CAGNNNCTG 3 cut(s) 103, 285, 1000
AoxI GGCC 4 cut(s) 417, 556, 1143, 1239
ApaLI GTGCAC 1 cut(s) 21
ApeKI GCWGC 4 cut(s) 88, 187, 377, 835
ApoI RAATTY 3 cut(s) 429, 500, 781
AsiGI ACCGGT 1 cut(s) 396
AspLEI GCGC 1 cut(s) 1069
AspS9I GGNCC 2 cut(s) 716, 1298
AsuC2I CCSGG 1 cut(s) 137
AsuHPI GGTGA 4 cut(s) 382, 448, 675, 735
AsuNHI GCTAGC 1 cut(s) 5
AvaII GGWCC 2 cut(s) 716, 1298
BaeGI GKGCMC 1 cut(s) 25
BalI TGGCCA 2 cut(s) 1145, 1241
BbsI GAAGAC 1 cut(s) 139
Bbv12I GWGCWC 1 cut(s) 25
BbvCI CCTCAGC 1 cut(s) 384
BbvI GCAGC 4 cut(s) 100, 199, 364, 847
BccI CCATC 3 cut(s) 631, 1025, 1276
BceAI ACGGC 1 cut(s) 354
BciT130I CCWGG 3 cut(s) 42, 937, 1056
BclI TGATCA 1 cut(s) 466
BcnI CCSGG 1 cut(s) 137
BcoDI GTCTC 3 cut(s) 236, 893, 971
BfaI CTAG 6 cut(s) 6, 456, 860, 947, 1031, 1250
BfmI CTRYAG 1 cut(s) 1155
BglII AGATCT 1 cut(s) 541
BisI GCNGC 4 cut(s) 89, 188, 378, 836
BlsI GCNGC 4 cut(s) 90, 189, 379, 837
Bme1390I CCNGG 4 cut(s) 42, 137, 937, 1056
Bme18I GGWCC 2 cut(s) 716, 1298
BmgT120I GGNCC 2 cut(s) 716, 1298
BmrFI CCNGG 4 cut(s) 42, 137, 937, 1056
BmrI ACTGGG 1 cut(s) 210
BmsI GCATC 3 cut(s) 746, 1105, 1283
BmtI GCTAGC 1 cut(s) 9
BmuI ACTGGG 1 cut(s) 210
BoxI GACNNNNGTC 1 cut(s) 288
BpiI GAAGAC 1 cut(s) 139
BpmI CTGGAG 3 cut(s) 307, 1077, 1086
Bpu10I CCTNAGC 2 cut(s) 384, 794
BpuEI CTTGAG 1 cut(s) 428
BpuMI CCSGG 1 cut(s) 137
Bsa29I ATCGAT 1 cut(s) 508
BsaJI CCNNGG 6 cut(s) 136, 559, 712, 757, 1166, 1301
BsaWI WCCGGW 1 cut(s) 396
Bsc4I CCNNNNNNNGG 3 cut(s) 285, 554, 1261
Bse118I RCCGGY 1 cut(s) 396
Bse1I ACTGG 4 cut(s) 205, 279, 290, 698
Bse3DI GCAATG 2 cut(s) 477, 1113
BseBI CCWGG 3 cut(s) 42, 937, 1056
BseCI ATCGAT 1 cut(s) 508
BseDI CCNNGG 6 cut(s) 136, 559, 712, 757, 1166, 1301
BseGI GGATG 4 cut(s) 582, 967, 1270, 1287
BseLI CCNNNNNNNGG 3 cut(s) 285, 554, 1261
BseMI GCAATG 2 cut(s) 477, 1113
BseMII CTCAG 5 cut(s) 95, 210, 231, 398, 992
BseNI ACTGG 4 cut(s) 205, 279, 290, 698
BseSI GKGCMC 1 cut(s) 25
BseXI GCAGC 4 cut(s) 100, 199, 364, 847
BseYI CCCAGC 1 cut(s) 9
BsgI GTGCAG 1 cut(s) 1251
Bsh1236I CGCG 1 cut(s) 1128
BshFI GGCC 4 cut(s) 419, 558, 1145, 1241
BshTI ACCGGT 1 cut(s) 396
BshVI ATCGAT 1 cut(s) 508
BsiHKAI GWGCWC 1 cut(s) 25
BsiSI CCGG 2 cut(s) 136, 397
BslFI GGGAC 1 cut(s) 869
BslI CCNNNNNNNGG 3 cut(s) 285, 554, 1261
BsmAI GTCTC 3 cut(s) 236, 893, 971
BsmFI GGGAC 1 cut(s) 869
BsmI GAATGC 2 cut(s) 565, 641
BsnI GGCC 4 cut(s) 419, 558, 1145, 1241
Bsp1286I GDGCHC 1 cut(s) 25
Bsp1407I TGTACA 1 cut(s) 346
Bsp143I GATC 8 cut(s) 466, 484, 505, 541, 575, 804, 1060, 1221
Bsp19I CCATGG 1 cut(s) 1166
BspACI CCGC 2 cut(s) 778, 1128
BspANI GGCC 4 cut(s) 419, 558, 1145, 1241
BspCNI CTCAG 5 cut(s) 96, 211, 232, 397, 993
BspDI ATCGAT 1 cut(s) 508
BspFNI CGCG 1 cut(s) 1128
BspHI TCATGA 1 cut(s) 807
BspOI GCTAGC 1 cut(s) 9
BspPI GGATC 2 cut(s) 583, 1216
BsrDI GCAATG 2 cut(s) 477, 1113
BsrFI RCCGGY 1 cut(s) 396
BsrGI TGTACA 1 cut(s) 346
BsrI ACTGG 4 cut(s) 205, 279, 290, 698
BssAI RCCGGY 1 cut(s) 396
BssECI CCNNGG 6 cut(s) 136, 559, 712, 757, 1166, 1301
BssMI GATC 8 cut(s) 466, 484, 505, 541, 575, 804, 1060, 1221
BssT1I CCWWGG 5 cut(s) 559, 712, 757, 1166, 1301
Bst2UI CCWGG 3 cut(s) 42, 937, 1056
Bst4CI ACNGT 3 cut(s) 247, 929, 1156
Bst6I CTCTTC 3 cut(s) 65, 865, 891
BstAPI GCANNNNNTGC 1 cut(s) 820
BstAUI TGTACA 1 cut(s) 346
BstC8I GCNNGC 2 cut(s) 7, 661
BstDEI CTNAG 6 cut(s) 104, 219, 240, 384, 794, 1001
BstDSI CCRYGG 1 cut(s) 1166
BstEII GGTNACC 1 cut(s) 108
BstF5I GGATG 4 cut(s) 582, 967, 1270, 1287
BstFNI CGCG 1 cut(s) 1128
BstHHI GCGC 1 cut(s) 1069
BstKTI GATC 8 cut(s) 469, 487, 508, 544, 578, 807, 1063, 1224
BstMAI GTCTC 3 cut(s) 236, 893, 971
BstMBI GATC 8 cut(s) 466, 484, 505, 541, 575, 804, 1060, 1221
BstMWI GCNNNNNNNGC 5 cut(s) 665, 820, 1075, 1238, 1312
BstNI CCWGG 3 cut(s) 42, 937, 1056
BstNSI RCATGY 1 cut(s) 1039
BstPAI GACNNNNGTC 1 cut(s) 288
BstPI GGTNACC 1 cut(s) 108
BstSCI CCNGG 4 cut(s) 40, 135, 935, 1054
BstSFI CTRYAG 1 cut(s) 1155
BstSLI GKGCMC 1 cut(s) 25
BstUI CGCG 1 cut(s) 1128
BstV1I GCAGC 4 cut(s) 100, 199, 364, 847
BstV2I GAAGAC 1 cut(s) 139
BstX2I RGATCY 3 cut(s) 541, 575, 1221
BstXI CCANNNNNNTGG 1 cut(s) 713
BstYI RGATCY 3 cut(s) 541, 575, 1221
Bsu15I ATCGAT 1 cut(s) 508
BsuRI GGCC 4 cut(s) 419, 558, 1145, 1241
BsuTUI ATCGAT 1 cut(s) 508
BtgI CCRYGG 1 cut(s) 1166
BtsCI GGATG 4 cut(s) 582, 967, 1270, 1287
BtsIMutI CAGTG 1 cut(s) 286
Cac8I GCNNGC 2 cut(s) 7, 661
CaiI CAGNNNCTG 3 cut(s) 103, 285, 1000
CciI TCATGA 1 cut(s) 807
CfoI GCGC 1 cut(s) 1069
Cfr10I RCCGGY 1 cut(s) 396
Cfr13I GGNCC 2 cut(s) 716, 1298
ClaI ATCGAT 1 cut(s) 508
CseI GACGC 1 cut(s) 401
Csp6I GTAC 1 cut(s) 347
CspAI ACCGGT 1 cut(s) 396
CviAII CATG 6 cut(s) 306, 808, 1036, 1064, 1167, 1317
CviQI GTAC 1 cut(s) 347
DdeI CTNAG 6 cut(s) 104, 219, 240, 384, 794, 1001
DpnI GATC 8 cut(s) 468, 486, 507, 543, 577, 806, 1062, 1223
DpnII GATC 8 cut(s) 466, 484, 505, 541, 575, 804, 1060, 1221
DraI TTTAAA 1 cut(s) 526
DraIII CACNNNGTG 1 cut(s) 263
EaeI YGGCCR 3 cut(s) 417, 1143, 1239
Eam1104I CTCTTC 3 cut(s) 65, 865, 891
EarI CTCTTC 3 cut(s) 65, 865, 891
Eco130I CCWWGG 5 cut(s) 559, 712, 757, 1166, 1301
Eco47I GGWCC 2 cut(s) 716, 1298
Eco57I CTGAAG 1 cut(s) 915
Eco91I GGTNACC 1 cut(s) 108
EcoO65I GGTNACC 1 cut(s) 108
EcoRII CCWGG 3 cut(s) 40, 935, 1054
EcoT14I CCWWGG 5 cut(s) 559, 712, 757, 1166, 1301
ErhI CCWWGG 5 cut(s) 559, 712, 757, 1166, 1301
FaeI CATG 6 cut(s) 309, 811, 1039, 1067, 1170, 1320
FaqI GGGAC 1 cut(s) 869
FatI CATG 6 cut(s) 305, 807, 1035, 1063, 1166, 1316
FauI CCCGC 1 cut(s) 1121
FbaI TGATCA 1 cut(s) 466
Fnu4HI GCNGC 4 cut(s) 89, 188, 378, 836
FokI GGATG 4 cut(s) 569, 974, 1277, 1294
Fsp4HI GCNGC 4 cut(s) 89, 188, 378, 836
FspBI CTAG 6 cut(s) 6, 456, 860, 947, 1031, 1250
GlaI GCGC 1 cut(s) 1068
GluI GCNGC 4 cut(s) 89, 188, 378, 836
GsaI CCCAGC 1 cut(s) 13
GsuI CTGGAG 3 cut(s) 307, 1077, 1086
HaeIII GGCC 4 cut(s) 419, 558, 1145, 1241
HapII CCGG 2 cut(s) 136, 397
HgaI GACGC 1 cut(s) 401
HhaI GCGC 1 cut(s) 1069
Hin1II CATG 6 cut(s) 309, 811, 1039, 1067, 1170, 1320
Hin6I GCGC 1 cut(s) 1067
HinP1I GCGC 1 cut(s) 1067
HincII GTYRAC 1 cut(s) 1041
HindII GTYRAC 1 cut(s) 1041
HpaII CCGG 2 cut(s) 136, 397
HphI GGTGA 4 cut(s) 382, 448, 675, 735
Hpy166II GTNNAC 2 cut(s) 23, 1041
Hpy188I TCNGA 6 cut(s) 105, 220, 315, 804, 1009, 1311
Hpy188III TCNNGA 4 cut(s) 407, 592, 603, 808
Hpy8I GTNNAC 2 cut(s) 23, 1041
Hpy99I CGWCG 1 cut(s) 340
HpyAV CCTTC 3 cut(s) 418, 1235, 1265
HpyCH4III ACNGT 3 cut(s) 247, 929, 1156
HpyCH4V TGCA 8 cut(s) 23, 443, 482, 737, 1078, 1118, 1232, 1274
HpyF10VI GCNNNNNNNGC 5 cut(s) 665, 820, 1075, 1238, 1312
HpyF3I CTNAG 6 cut(s) 104, 219, 240, 384, 794, 1001
Hsp92II CATG 6 cut(s) 309, 811, 1039, 1067, 1170, 1320
HspAI GCGC 1 cut(s) 1067
Ksp22I TGATCA 1 cut(s) 466
Kzo9I GATC 8 cut(s) 466, 484, 505, 541, 575, 804, 1060, 1221
LmnI GCTCC 3 cut(s) 374, 832, 856
Lsp1109I GCAGC 4 cut(s) 100, 199, 364, 847
LweI GCATC 3 cut(s) 746, 1105, 1283
MaeI CTAG 6 cut(s) 6, 456, 860, 947, 1031, 1250
MaeIII GTNAC 3 cut(s) 108, 152, 281
MalI GATC 8 cut(s) 468, 486, 507, 543, 577, 806, 1062, 1223
MboI GATC 8 cut(s) 466, 484, 505, 541, 575, 804, 1060, 1221
MfeI CAATTG 2 cut(s) 534, 969
MflI RGATCY 3 cut(s) 541, 575, 1221
MhlI GDGCHC 1 cut(s) 25
MlsI TGGCCA 2 cut(s) 1145, 1241
MluCI AATT 8 cut(s) 115, 429, 494, 500, 534, 720, 781, 969
MluNI TGGCCA 2 cut(s) 1145, 1241
MlyI GAGTC 4 cut(s) 153, 298, 895, 1181
MmeI TCCRAC 2 cut(s) 248, 352
Mox20I TGGCCA 2 cut(s) 1145, 1241
MscI TGGCCA 2 cut(s) 1145, 1241
MseI TTAA 1 cut(s) 525
Msp20I TGGCCA 2 cut(s) 1145, 1241
MspI CCGG 2 cut(s) 136, 397
MspR9I CCNGG 4 cut(s) 42, 137, 937, 1056
MunI CAATTG 2 cut(s) 534, 969
Mva1269I GAATGC 2 cut(s) 565, 641
MvaI CCWGG 3 cut(s) 42, 937, 1056
MvnI CGCG 1 cut(s) 1128
MwoI GCNNNNNNNGC 5 cut(s) 665, 820, 1075, 1238, 1312
NciI CCSGG 1 cut(s) 137
NcoI CCATGG 1 cut(s) 1166
NdeII GATC 8 cut(s) 466, 484, 505, 541, 575, 804, 1060, 1221
NheI GCTAGC 1 cut(s) 5
NlaIII CATG 6 cut(s) 309, 811, 1039, 1067, 1170, 1320
NmeAIII GCCGAG 1 cut(s) 445
NmuCI GTSAC 1 cut(s) 281
NspI RCATGY 1 cut(s) 1039
PagI TCATGA 1 cut(s) 807
PciI ACATGT 1 cut(s) 1035
PctI GAATGC 2 cut(s) 565, 641
PfeI GAWTC 6 cut(s) 82, 182, 275, 940, 1010, 1211
PflMI CCANNNNNTGG 1 cut(s) 285
PfoI TCCNGGA 1 cut(s) 935
PinAI ACCGGT 1 cut(s) 396
PkrI GCNGC 4 cut(s) 90, 189, 379, 837
PleI GAGTC 4 cut(s) 152, 297, 895, 1180
PpsI GAGTC 4 cut(s) 152, 297, 895, 1180
PscI ACATGT 1 cut(s) 1035
PshAI GACNNNNGTC 1 cut(s) 288
PsiI TTATAA 1 cut(s) 914
Psp6I CCWGG 3 cut(s) 40, 935, 1054
PspEI GGTNACC 1 cut(s) 108
PspFI CCCAGC 1 cut(s) 9
PspGI CCWGG 3 cut(s) 40, 935, 1054
PspPI GGNCC 2 cut(s) 716, 1298
PstNI CAGNNNCTG 3 cut(s) 103, 285, 1000
PsuI RGATCY 3 cut(s) 541, 575, 1221
RsaI GTAC 1 cut(s) 348
RsaNI GTAC 1 cut(s) 347
SaqAI TTAA 1 cut(s) 525
SatI GCNGC 4 cut(s) 89, 188, 378, 836
Sau3AI GATC 8 cut(s) 466, 484, 505, 541, 575, 804, 1060, 1221
Sau96I GGNCC 2 cut(s) 716, 1298
SchI GAGTC 4 cut(s) 153, 298, 895, 1181
ScrFI CCNGG 4 cut(s) 42, 137, 937, 1056
SduI GDGCHC 1 cut(s) 25
SfaNI GCATC 3 cut(s) 746, 1105, 1283
SfcI CTRYAG 1 cut(s) 1155
SinI GGWCC 2 cut(s) 716, 1298
SmlI CTYRAG 1 cut(s) 407
SmoI CTYRAG 1 cut(s) 407
Sse9I AATT 8 cut(s) 115, 429, 494, 500, 534, 720, 781, 969
SsiI CCGC 2 cut(s) 778, 1128
SspI AATATT 1 cut(s) 987
SspMI CTAG 6 cut(s) 6, 456, 860, 947, 1031, 1250
StyD4I CCNGG 4 cut(s) 40, 135, 935, 1054
StyI CCWWGG 5 cut(s) 559, 712, 757, 1166, 1301
TaaI ACNGT 3 cut(s) 247, 929, 1156
TaqI TCGA 1 cut(s) 508
TasI AATT 8 cut(s) 115, 429, 494, 500, 534, 720, 781, 969
TatI WGTACW 1 cut(s) 346
TfiI GAWTC 6 cut(s) 82, 182, 275, 940, 1010, 1211
Tru1I TTAA 1 cut(s) 525
Tru9I TTAA 1 cut(s) 525
TscAI CASTG 1 cut(s) 286
TseFI GTSAC 1 cut(s) 281
TseI GCWGC 4 cut(s) 88, 187, 377, 835
Tsp45I GTSAC 1 cut(s) 281
TspDTI ATGAA 5 cut(s) 20, 38, 180, 466, 824
TspRI CASTG 1 cut(s) 286
Van91I CCANNNNNTGG 1 cut(s) 285
VneI GTGCAC 1 cut(s) 21
VpaK11BI GGWCC 2 cut(s) 716, 1298
XapI RAATTY 3 cut(s) 429, 500, 781
XceI RCATGY 1 cut(s) 1039
XcmI CCANNNNNNNNNTGG 1 cut(s) 593
XspI CTAG 6 cut(s) 6, 456, 860, 947, 1031, 1250
Using CommOnly database (standard laboratory enzymes). Scanned on CDS sequence.