Rh7BG259400

Vacuolar-processing enzyme-like

Design Primers CRISPR Design

Basic Information

Type: gene
Biological Identity
rosa_samantha
Chr7B
OG0001085
Physical Location & Seq
Forward (+)
23611880 .. 23617194
5315 bp
Loading structure...
UTR
Exon/CDS
Intron
Rh7BG259400.1

Sequence Viewer

Length: 1509 bp
ATGAGAGGGCAGTACTGTTCCAGTGGAGTATTTTTGGTATTGCTTACCATCTTTGGTGTGAGCAGTGCTCAACTTCCCAAGTTGATGCCACAAGAATTTCCGAAGGATTTCATTATTCATGTTGATAGGGGCTCTAGCAACAGTGACACAAATGGAACAAGATGGGCAGTTCTAGTTGCTGGGTCTATGGGTTATGGAAACTATAGGCATCAGGCTGATGTGTGTCATGCATATCAAATTTTGAAGAAAGGTGGACTGCAAGATGAAAACATCATTGTTTTCATGTACGATGACATTGCAAACAACCCTGAGAATCCAAGGCCTGGTGTCATCATCAACAAGCCCGACGGCGATGACGTCTACCATGGAGTTCCCAAGGATTACGCAGGAGAGAATGCTACTGCTGCTAATCTTTATGCTGTCATTCTTGGAGATAAATCTAATCTCTCTGGGGGTAGTGGCAAGGTTCTGAACAGCGGGCCAAACGACAATGTCTTTATCTATTACACCGACCATGGCGGGCCTGGGATAGTCGGGATGCCTGAAGGCGATTTCGTCTATGCCAATGATCTGGTTGATGTTCTGATAAAGAAGCATGAAGCTAATGCATACAAAAATATGGTATTTTACCTTGAAGCTTGTGAATCCGGGAGTATGTTCGAGGGTCTACTGCCGACAAATATAAGCATCTATGCAACCACAGCATCAAATGCAACTGAGAATAGTTGGGGAACGTACTGCCCTGATATGACTCCTCCCCCACCTCCTGAATATGACACTTGCTTGGGTGATTTGTATAGTATTTCTTGGATGGAGGACTGTGACTTACAAGATTTGCGCCAAGAGTCTCTGGAAGAGCAATATCAATCGGTTAAATGGAGAACAGTAAACAATACTTATGGTTCTCATGTTATGCAATATGGAAATATGTATCAAACCTCAGATTTAGTATCCTTTTACTTGGGTGCAAATGATAACTCTTCCTCCATTGAAAGCAAATCGCCAGAGTCAATCTTAGAAGTTGTTGACAATCGCGATGCAGATCTTCTTCATCTTTGGCACAAGTTCCAACGCGCACCCGCTGGCTCTGAAATGAAGCAGGAAGCTAAAAGGGTCCTAGATGATGAAATTTCAAGTAGGCAGCGCATTGACCACAACATAAACCAAATCGAACAACTTGTGTTGGGATATTCAAGTAACTCTGTTCGTCCTGGGGGGCAAGCTCTTGTTGATGATTGGGACTGCTTTAAGACAGTCGTGAAAACTTATGAGAAACATTGTGGGTCGTTGTCAAATTATGGTATGAAACACACACGAGTTTTTGCCAACATGTGCAATGCTGGGATCAGCGTGGAGCAAATCGAGTTGCACAAGGTTAAAAGTTGCTACTTTAGAATAGGGTCTTGGATCGACTTGGATCCCCCGGTTTTCACCCTAGATGTAGGGTTCCGGGATGGCAGACCGCCGCGAACGTTGAACATCACTCATCGGCAACATTTTAAAACGTGA
Functional Annotation
Gene Ontology
Molecular Function
Biological Process
Cellular Component
No ontology terms assigned.
KEGG Pathways
Metabolic & Signaling
ko04142 ko04612 map04142 map04612
Pfam Domains
Protein Families
Peptidase_C13

Protein Analysis

502

Amino Acids

56.06

Weight (kDa)

5.1

Isoelectric Point (pI)

44.23

Instability Index
Protein Domains (Pfam)
Domain Name Pfam ID Position E-value Description
Peptidase_C13 PF01650 54 - 322 3.5e-114 Peptidase C13 family
Legum_prodom PF20985 373 - 462 2.8e-31 Legumain, prodomain
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)
AatII GACGTC 1 cut(s) 360
AccB7I CCANNNNNTGG 1 cut(s) 323
AccI GTMKAC 2 cut(s) 360, 667
AccII CGCG 3 cut(s) 1035, 1074, 1468
AciI CCGC 5 cut(s) 477, 519, 1080, 1463, 1466
AclI AACGTT 1 cut(s) 1472
AclWI GGATC 4 cut(s) 1352, 1412, 1415, 1425
AcsI RAATTY 3 cut(s) 95, 237, 1128
AcuI CTGAAG 1 cut(s) 564
AcyI GRCGYC 1 cut(s) 357
AfaI GTAC 3 cut(s) 14, 287, 737
AfiI CCNNNNNNNGG 1 cut(s) 323
AflIII ACRYGT 1 cut(s) 1329
AgsI TTSAA 6 cut(s) 244, 635, 992, 1134, 1194, 1477
AjnI CCWGG 3 cut(s) 322, 523, 1210
AjuI GAANNNNNNNTTGG 2 cut(s) 1387, 1419
AluBI AGCT 4 cut(s) 602, 638, 1106, 1223
AluI AGCT 4 cut(s) 602, 638, 1106, 1223
Alw21I GWGCWC 1 cut(s) 70
Alw26I GTCTC 1 cut(s) 852
AlwI GGATC 4 cut(s) 1352, 1412, 1415, 1425
AoxI GGCC 3 cut(s) 320, 479, 521
ApeKI GCWGC 2 cut(s) 404, 1141
ApoI RAATTY 3 cut(s) 95, 237, 1128
Asp700I GAANNNNTTC 1 cut(s) 107
AspLEI GCGC 3 cut(s) 840, 1076, 1146
AspS9I GGNCC 3 cut(s) 479, 521, 1114
AsuC2I CCSGG 3 cut(s) 649, 1424, 1451
AsuHPI GGTGA 2 cut(s) 800, 1423
AvaII GGWCC 1 cut(s) 1114
BamHI GGATCC 1 cut(s) 1417
BanII GRGCYC 1 cut(s) 134
BauI CACGAG 1 cut(s) 1314
Bbv12I GWGCWC 1 cut(s) 70
BbvI GCAGC 2 cut(s) 391, 1153
BccI CCATC 4 cut(s) 56, 156, 805, 1448
BceAI ACGGC 1 cut(s) 364
BcgI CGANNNNNNTGC 2 cut(s) 278, 312
BciT130I CCWGG 3 cut(s) 324, 525, 1212
BciVI GTATCC 1 cut(s) 961
BcnI CCSGG 3 cut(s) 649, 1424, 1451
BcoDI GTCTC 1 cut(s) 852
BfaI CTAG 4 cut(s) 135, 173, 1118, 1436
BfmI CTRYAG 1 cut(s) 202
BfuI GTATCC 1 cut(s) 961
BglII AGATCT 1 cut(s) 1042
BisI GCNGC 3 cut(s) 405, 1142, 1466
BlsI GCNGC 3 cut(s) 406, 1143, 1467
BmcAI AGTACT 1 cut(s) 14
Bme1390I CCNGG 6 cut(s) 324, 525, 649, 1212, 1424, 1451
Bme18I GGWCC 1 cut(s) 1114
BmgT120I GGNCC 3 cut(s) 479, 521, 1114
BmiI GGNNCC 3 cut(s) 1115, 1419, 1448
BmrFI CCNGG 6 cut(s) 324, 525, 649, 1212, 1424, 1451
BmsI GCATC 6 cut(s) 75, 217, 528, 696, 713, 1027
BplI GAGNNNNNCTC 2 cut(s) 52, 84
BpuMI CCSGG 3 cut(s) 649, 1424, 1451
BsaBI GATNNNNATC 1 cut(s) 1041
BsaHI GRCGYC 1 cut(s) 357
BsaJI CCNNGG 7 cut(s) 317, 364, 375, 514, 524, 1211, 1422
BsaXI ACNNNNNCTCC 2 cut(s) 968, 998
Bsc4I CCNNNNNNNGG 1 cut(s) 323
Bse1I ACTGG 1 cut(s) 21
Bse3DI GCAATG 2 cut(s) 294, 1342
Bse8I GATNNNNATC 1 cut(s) 1041
BseBI CCWGG 3 cut(s) 324, 525, 1212
BseDI CCNNGG 7 cut(s) 317, 364, 375, 514, 524, 1211, 1422
BseGI GGATG 3 cut(s) 543, 816, 1459
BseJI GATNNNNATC 1 cut(s) 1041
BseLI CCNNNNNNNGG 1 cut(s) 323
BseMI GCAATG 2 cut(s) 294, 1342
BseMII CTCAG 3 cut(s) 300, 708, 954
BseNI ACTGG 1 cut(s) 21
BseRI GAGGAG 1 cut(s) 744
BseXI GCAGC 2 cut(s) 391, 1153
BseYI CCCAGC 2 cut(s) 179, 1340
Bsh1236I CGCG 3 cut(s) 1035, 1074, 1468
BshFI GGCC 3 cut(s) 322, 481, 523
BsiHKAI GWGCWC 1 cut(s) 70
BsiSI CCGG 3 cut(s) 648, 1424, 1450
BslFI GGGAC 1 cut(s) 1253
BslI CCNNNNNNNGG 1 cut(s) 323
BsmAI GTCTC 1 cut(s) 852
BsmFI GGGAC 1 cut(s) 1253
BsmI GAATGC 1 cut(s) 400
BsnI GGCC 3 cut(s) 322, 481, 523
Bsp1286I GDGCHC 2 cut(s) 70, 134
Bsp143I GATC 5 cut(s) 568, 1042, 1344, 1407, 1417
Bsp19I CCATGG 2 cut(s) 364, 514
Bsp68I TCGCGA 1 cut(s) 1035
BspACI CCGC 5 cut(s) 477, 519, 1080, 1463, 1466
BspANI GGCC 3 cut(s) 322, 481, 523
BspCNI CTCAG 3 cut(s) 301, 709, 953
BspFNI CGCG 3 cut(s) 1035, 1074, 1468
BspLI GGNNCC 3 cut(s) 1115, 1419, 1448
BspPI GGATC 4 cut(s) 1352, 1412, 1415, 1425
BspQI GCTCTTC 1 cut(s) 849
BsrDI GCAATG 2 cut(s) 294, 1342
BsrI ACTGG 1 cut(s) 21
BssECI CCNNGG 7 cut(s) 317, 364, 375, 514, 524, 1211, 1422
BssMI GATC 5 cut(s) 568, 1042, 1344, 1407, 1417
BssNI GRCGYC 1 cut(s) 357
BssSI CACGAG 1 cut(s) 1314
BssT1I CCWWGG 4 cut(s) 317, 364, 375, 514
Bst2BI CACGAG 1 cut(s) 1314
Bst2UI CCWGG 3 cut(s) 324, 525, 1212
Bst4CI ACNGT 5 cut(s) 17, 143, 821, 886, 1255
Bst6I CTCTTC 2 cut(s) 849, 985
BstACI GRCGYC 1 cut(s) 357
BstAPI GCANNNNNTGC 1 cut(s) 710
BstC8I GCNNGC 4 cut(s) 479, 521, 1084, 1221
BstDEI CTNAG 4 cut(s) 309, 717, 940, 1015
BstDSI CCRYGG 2 cut(s) 364, 514
BstF5I GGATG 3 cut(s) 543, 816, 1459
BstFNI CGCG 3 cut(s) 1035, 1074, 1468
BstHHI GCGC 3 cut(s) 840, 1076, 1146
BstKTI GATC 5 cut(s) 571, 1045, 1347, 1410, 1420
BstMAI GTCTC 1 cut(s) 852
BstMBI GATC 5 cut(s) 568, 1042, 1344, 1407, 1417
BstMWI GCNNNNNNNGC 3 cut(s) 404, 701, 710
BstNI CCWGG 3 cut(s) 324, 525, 1212
BstNSI RCATGY 1 cut(s) 1333
BstSCI CCNGG 6 cut(s) 322, 523, 647, 1210, 1422, 1449
BstSFI CTRYAG 1 cut(s) 202
BstUI CGCG 3 cut(s) 1035, 1074, 1468
BstV1I GCAGC 2 cut(s) 391, 1153
BstX2I RGATCY 2 cut(s) 1042, 1417
BstXI CCANNNNNNTGG 1 cut(s) 571
BstYI RGATCY 2 cut(s) 1042, 1417
BsuI GTATCC 1 cut(s) 961
BsuRI GGCC 3 cut(s) 322, 481, 523
BtgI CCRYGG 2 cut(s) 364, 514
BtgZI GCGATG 2 cut(s) 366, 1050
BtsCI GGATG 3 cut(s) 543, 816, 1459
BtsI GCAGTG 1 cut(s) 70
BtsIMutI CAGTG 3 cut(s) 28, 70, 148
BtuMI TCGCGA 1 cut(s) 1035
Cac8I GCNNGC 4 cut(s) 479, 521, 1084, 1221
CfoI GCGC 3 cut(s) 840, 1076, 1146
Cfr13I GGNCC 3 cut(s) 479, 521, 1114
Csp6I GTAC 3 cut(s) 13, 286, 736
CviAII CATG 8 cut(s) 119, 227, 283, 365, 515, 596, 908, 1330
CviQI GTAC 3 cut(s) 13, 286, 736
DdeI CTNAG 4 cut(s) 309, 717, 940, 1015
DpnI GATC 5 cut(s) 570, 1044, 1346, 1409, 1419
DpnII GATC 5 cut(s) 568, 1042, 1344, 1407, 1417
DraI TTTAAA 1 cut(s) 1501
Eam1104I CTCTTC 2 cut(s) 849, 985
EarI CTCTTC 2 cut(s) 849, 985
Eco130I CCWWGG 4 cut(s) 317, 364, 375, 514
Eco147I AGGCCT 1 cut(s) 322
Eco24I GRGCYC 1 cut(s) 134
Eco47I GGWCC 1 cut(s) 1114
Eco57I CTGAAG 1 cut(s) 564
EcoO109I RGGNCCY 1 cut(s) 1114
EcoRII CCWGG 3 cut(s) 322, 523, 1210
EcoT14I CCWWGG 4 cut(s) 317, 364, 375, 514
EcoT22I ATGCAT 2 cut(s) 232, 610
EcoT38I GRGCYC 1 cut(s) 134
ErhI CCWWGG 4 cut(s) 317, 364, 375, 514
FaeI CATG 8 cut(s) 122, 230, 286, 368, 518, 599, 911, 1333
FaqI GGGAC 1 cut(s) 1253
FatI CATG 8 cut(s) 118, 226, 282, 364, 514, 595, 907, 1329
FauI CCCGC 3 cut(s) 470, 512, 1087
FblI GTMKAC 2 cut(s) 360, 667
Fnu4HI GCNGC 3 cut(s) 405, 1142, 1466
FokI GGATG 3 cut(s) 550, 823, 1466
FriOI GRGCYC 1 cut(s) 134
Fsp4HI GCNGC 3 cut(s) 405, 1142, 1466
FspBI CTAG 4 cut(s) 135, 173, 1118, 1436
GlaI GCGC 3 cut(s) 839, 1075, 1145
GluI GCNGC 3 cut(s) 405, 1142, 1466
GsaI CCCAGC 2 cut(s) 183, 1344
HaeIII GGCC 3 cut(s) 322, 481, 523
HapII CCGG 3 cut(s) 648, 1424, 1450
HhaI GCGC 3 cut(s) 840, 1076, 1146
Hin1I GRCGYC 1 cut(s) 357
Hin1II CATG 8 cut(s) 122, 230, 286, 368, 518, 599, 911, 1333
Hin6I GCGC 3 cut(s) 838, 1074, 1144
HinP1I GCGC 3 cut(s) 838, 1074, 1144
HincII GTYRAC 1 cut(s) 1027
HindII GTYRAC 1 cut(s) 1027
HindIII AAGCTT 1 cut(s) 636
HinfI GANTC 5 cut(s) 313, 644, 751, 845, 1007
HpaII CCGG 3 cut(s) 648, 1424, 1450
HphI GGTGA 2 cut(s) 800, 1423
Hpy166II GTNNAC 5 cut(s) 254, 361, 668, 889, 1027
Hpy188I TCNGA 5 cut(s) 102, 471, 585, 943, 1090
Hpy188III TCNNGA 5 cut(s) 535, 767, 851, 1034, 1258
Hpy8I GTNNAC 5 cut(s) 254, 361, 668, 889, 1027
Hpy99I CGWCG 1 cut(s) 350
HpyAV CCTTC 2 cut(s) 97, 539
HpyCH4III ACNGT 5 cut(s) 17, 143, 821, 886, 1255
HpyCH4IV ACGT 4 cut(s) 357, 734, 1472, 1505
HpyF10VI GCNNNNNNNGC 3 cut(s) 404, 701, 710
HpyF3I CTNAG 4 cut(s) 309, 717, 940, 1015
HpySE526I ACGT 4 cut(s) 357, 734, 1472, 1505
Hsp92I GRCGYC 1 cut(s) 357
Hsp92II CATG 8 cut(s) 122, 230, 286, 368, 518, 599, 911, 1333
HspAI GCGC 3 cut(s) 838, 1074, 1144
Kzo9I GATC 5 cut(s) 568, 1042, 1344, 1407, 1417
LguI GCTCTTC 1 cut(s) 849
LmnI GCTCC 1 cut(s) 1354
Lsp1109I GCAGC 2 cut(s) 391, 1153
LweI GCATC 6 cut(s) 75, 217, 528, 696, 713, 1027
MaeI CTAG 4 cut(s) 135, 173, 1118, 1436
MaeII ACGT 4 cut(s) 357, 734, 1472, 1505
MaeIII GTNAC 3 cut(s) 143, 821, 1196
MalI GATC 5 cut(s) 570, 1044, 1346, 1409, 1419
MboI GATC 5 cut(s) 568, 1042, 1344, 1407, 1417
MboII GAAGA 5 cut(s) 256, 866, 972, 1037, 1040
MflI RGATCY 2 cut(s) 1042, 1417
MhlI GDGCHC 2 cut(s) 70, 134
MluCI AATT 4 cut(s) 95, 237, 1128, 1294
MlyI GAGTC 3 cut(s) 745, 854, 1016
MmeI TCCRAC 1 cut(s) 1093
MnlI CCTC 6 cut(s) 655, 765, 774, 808, 949, 994
Mph1103I ATGCAT 2 cut(s) 232, 610
MroXI GAANNNNTTC 1 cut(s) 107
MseI TTAA 4 cut(s) 873, 1248, 1377, 1500
MspA1I CMGCKG 2 cut(s) 477, 1082
MspI CCGG 3 cut(s) 648, 1424, 1450
MspR9I CCNGG 6 cut(s) 324, 525, 649, 1212, 1424, 1451
Mva1269I GAATGC 1 cut(s) 400
MvaI CCWGG 3 cut(s) 324, 525, 1212
MvnI CGCG 3 cut(s) 1035, 1074, 1468
MwoI GCNNNNNNNGC 3 cut(s) 404, 701, 710
NciI CCSGG 3 cut(s) 649, 1424, 1451
NcoI CCATGG 2 cut(s) 364, 514
NdeII GATC 5 cut(s) 568, 1042, 1344, 1407, 1417
NlaIII CATG 8 cut(s) 122, 230, 286, 368, 518, 599, 911, 1333
NlaIV GGNNCC 3 cut(s) 1115, 1419, 1448
NmuCI GTSAC 2 cut(s) 143, 821
NruI TCGCGA 1 cut(s) 1035
NsiI ATGCAT 2 cut(s) 232, 610
NspI RCATGY 1 cut(s) 1333
PceI AGGCCT 1 cut(s) 322
PciI ACATGT 1 cut(s) 1329
PciSI GCTCTTC 1 cut(s) 849
PcsI WCGNNNNNNNCGW 1 cut(s) 354
PctI GAATGC 1 cut(s) 400
PdmI GAANNNNTTC 1 cut(s) 107
PfeI GAWTC 2 cut(s) 313, 644
PflFI GACNNNGTC 1 cut(s) 491
PflMI CCANNNNNTGG 1 cut(s) 323
PfoI TCCNGGA 2 cut(s) 647, 1449
PkrI GCNGC 3 cut(s) 406, 1143, 1467
PleI GAGTC 3 cut(s) 745, 853, 1015
PpsI GAGTC 3 cut(s) 745, 853, 1015
PpuMI RGGWCCY 1 cut(s) 1114
PscI ACATGT 1 cut(s) 1329
Psp1406I AACGTT 1 cut(s) 1472
Psp5II RGGWCCY 1 cut(s) 1114
Psp6I CCWGG 3 cut(s) 322, 523, 1210
PspFI CCCAGC 2 cut(s) 179, 1340
PspGI CCWGG 3 cut(s) 322, 523, 1210
PspN4I GGNNCC 3 cut(s) 1115, 1419, 1448
PspPI GGNCC 3 cut(s) 479, 521, 1114
PspPPI RGGWCCY 1 cut(s) 1114
PsuI RGATCY 2 cut(s) 1042, 1417
PsyI GACNNNGTC 1 cut(s) 491
RruI TCGCGA 1 cut(s) 1035
RsaI GTAC 3 cut(s) 14, 287, 737
RsaNI GTAC 3 cut(s) 13, 286, 736
SapI GCTCTTC 1 cut(s) 849
SaqAI TTAA 4 cut(s) 873, 1248, 1377, 1500
SatI GCNGC 3 cut(s) 405, 1142, 1466
Sau3AI GATC 5 cut(s) 568, 1042, 1344, 1407, 1417
Sau96I GGNCC 3 cut(s) 479, 521, 1114
ScaI AGTACT 1 cut(s) 14
SchI GAGTC 3 cut(s) 745, 854, 1016
ScrFI CCNGG 6 cut(s) 324, 525, 649, 1212, 1424, 1451
SduI GDGCHC 2 cut(s) 70, 134
SfaNI GCATC 6 cut(s) 75, 217, 528, 696, 713, 1027
SfcI CTRYAG 1 cut(s) 202
SinI GGWCC 1 cut(s) 1114
Sse9I AATT 4 cut(s) 95, 237, 1128, 1294
SseBI AGGCCT 1 cut(s) 322
SsiI CCGC 5 cut(s) 477, 519, 1080, 1463, 1466
SspMI CTAG 4 cut(s) 135, 173, 1118, 1436
StuI AGGCCT 1 cut(s) 322
StyD4I CCNGG 6 cut(s) 322, 523, 647, 1210, 1422, 1449
StyI CCWWGG 4 cut(s) 317, 364, 375, 514
TaaI ACNGT 5 cut(s) 17, 143, 821, 886, 1255
TaiI ACGT 4 cut(s) 360, 737, 1475, 1508
TaqI TCGA 4 cut(s) 660, 1170, 1362, 1410
TasI AATT 4 cut(s) 95, 237, 1128, 1294
TatI WGTACW 1 cut(s) 12
TauI GCSGC 1 cut(s) 1468
TfiI GAWTC 2 cut(s) 313, 644
Tru1I TTAA 4 cut(s) 873, 1248, 1377, 1500
Tru9I TTAA 4 cut(s) 873, 1248, 1377, 1500
TscAI CASTG 3 cut(s) 28, 70, 148
TseFI GTSAC 2 cut(s) 143, 821
TseI GCWGC 2 cut(s) 404, 1141
Tsp45I GTSAC 2 cut(s) 143, 821
TspDTI ATGAA 9 cut(s) 100, 107, 271, 279, 612, 1040, 1109, 1140, 1319
TspRI CASTG 3 cut(s) 28, 70, 148
Tth111I GACNNNGTC 1 cut(s) 491
Van91I CCANNNNNTGG 1 cut(s) 323
VpaK11BI GGWCC 1 cut(s) 1114
XapI RAATTY 3 cut(s) 95, 237, 1128
XceI RCATGY 1 cut(s) 1333
XcmI CCANNNNNNNNNTGG 1 cut(s) 521
XmiI GTMKAC 2 cut(s) 360, 667
XmnI GAANNNNTTC 1 cut(s) 107
XspI CTAG 4 cut(s) 135, 173, 1118, 1436
ZraI GACGTC 1 cut(s) 358
ZrmI AGTACT 1 cut(s) 14
Zsp2I ATGCAT 2 cut(s) 232, 610
Using CommOnly database (standard laboratory enzymes). Scanned on CDS sequence.