RLG00000024229

expansin-A9-like

Design Primers CRISPR Design

Basic Information

Type: gene
Biological Identity
rosa_laevigata
Chr5
OG0004238
Physical Location & Seq
Forward (+)
33323312 .. 33326042
2731 bp
Loading structure...
UTR
Exon/CDS
Intron
RLM00000024229

Sequence Viewer

Length: 1443 bp
ATGGCACCAGTCTTTCACGCCCTCTTGGGTTCATTACTACTCGTTCTTCCTTTGCTGCCTCATTATGTTCATGCAAATGCACGGAAAAACGTTGGTAAATATACAGGGGCTGGTTCACGCAGTCATGACAAGCTTCATGTAGTGGGTCGCTCCCACAGGCCAAGGTTCGCTACCGGCCCTTGGAAGCATGCTCACGCCACATTCTACGAAGGAGGCTCCGGTACATTTGGGGGAGCTTGCGGCTACGAAGATGTAGTTAAAGAAGGATATGGGTTGGACACAGCAGCATTGAGCAATGCATTGTTCAACAATGGGCAGGCTTGTGGTTCATGTTATGAAATCAAATGTGTGGAAAATCCTCAGTGGTGCAAACCAGGACAGAAGTCACTTTTTGTTACGGCCACCAACAATTGCCCCCCAAATAACCAGCAGTCCGCTGATAATGGAGGATGGTGCAATCCACCTCGTGAGCATTTCGATATAGCCAAGCCTTCATTCAGCAAAATTGCCGAGTACAAGGCCGGCATTACTCCTGTCATGTACCGCAGGGTTCCTTGCCAAAAGCAAGGAGGCATAAAGTTTACAATAACAGGGAATCCTTACTTCAACCAAGTGTTGGTCTGGAACGTCGCAGGTGCCGGAGATGTGACCAGCGTTGAAGTAAAGGGCCACAGGAACCTGAAATGGACACCGCTGAAGCGCCTTTGGGGTCAGAAGTGGGTGACTGACCAGAAAATGGTTGGTGAGAAACTGACCTTCCGAGTTAGGACGAGTGATGGAAGATCCTCAACCTCATGGCATATAATCCCTAACGACTGGCAGTTTGGCCAGACCTTCCAAGGCAAGAATTTCCCTCGACTCTATCTCCATCGTGGACCCGCCTCCTCCGCCGCCGTTAAATCTGCTCAGCCCGTCGCCGAACTAGGGAGAGACCATTGGGCCATGGACCGCCCCGGAAAGATTGGGATCCGAAACGGGGAAGGAGTTGACGGCGAAAGTGGGCTGACCGGGCTTGATTTTGCTGGGAGGGATGAACATGAGGCCAACTTGGCGGCGAGCGGCTGGGGTTGGGACGACGACGTATTGGGAGAAGGTGAAGTTGATGGTGGAGTTGAGGATGGAGAAGGAGGGGAGCTTAACGGAGTTGATGGAGATTTTGGGGTCTTTGGGTCAAACCCACCAAACCCAGAAACAATTTTTCAGAATCTAGAAGAACCCTTCTCTGCATTCTACAACATGTCTCCTCTTCTCCTAAATCTCTCATTAATGAGAACCCTCTCCAGCTGGTCTACCCTTTGCGACAACGACCCTAGTAGCCGGGGGTGTTGTTTGAATCAAAAAGTTTGGGGACAGGGTCGTGTGGTTGTGGTTGTGGCTATGGTGCTGATCGATGATTGCTCACCGACTCTATTGGACTATTGTTATGCTGCGCCGCCGCCTTGA
Functional Annotation
Gene Ontology
Molecular Function
Biological Process
Cellular Component
GO:0005575 GO:0005618 GO:0005623 GO:0006949 GO:0008150 GO:0009653 GO:0009987 GO:0016020 GO:0030312 GO:0032502 GO:0044464 GO:0048646 GO:0048856 GO:0071944
KEGG Pathways
Metabolic & Signaling
No pathways identified.
Pfam Domains
Protein Families
DPBB_1 Pollen_allerg_1

Protein Analysis

481

Amino Acids

52.29

Weight (kDa)

6.54

Isoelectric Point (pI)

30.12

Instability Index
Protein Domains (Pfam)
Domain Name Pfam ID Position E-value Description
EXPB1_D1 PF22514 66 - 188 1.7e-08 EXPB1-like domain 1
DPBB_1 PF03330 94 - 181 3.4e-20 Lytic transglycolase
Expansin_C PF01357 192 - 270 2.5e-23 Expansin C-terminal 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) 623
Acc36I ACCTGC 1 cut(s) 623
AccB1I GGYRCC 2 cut(s) 4, 635
AccB7I CCANNNNNTGG 2 cut(s) 616, 736
AccBSI CCGCTC 1 cut(s) 1059
AccI GTMKAC 1 cut(s) 1289
AclI AACGTT 1 cut(s) 90
AclWI GGATC 3 cut(s) 777, 961, 974
AcoI YGGCCR 2 cut(s) 399, 826
AcsI RAATTY 1 cut(s) 847
AcuI CTGAAG 1 cut(s) 716
AdeI CACNNNGTG 1 cut(s) 467
AfaI GTAC 3 cut(s) 223, 515, 542
AfiI CCNNNNNNNGG 5 cut(s) 180, 616, 736, 924, 976
AflIII ACRYGT 1 cut(s) 1236
AgsI TTSAA 4 cut(s) 307, 607, 659, 1333
AjnI CCWGG 1 cut(s) 373
AjuI GAANNNNNNNTTGG 2 cut(s) 479, 511
AluBI AGCT 4 cut(s) 133, 236, 1135, 1284
AluI AGCT 4 cut(s) 133, 236, 1135, 1284
Alw26I GTCTC 2 cut(s) 924, 1245
AlwI GGATC 3 cut(s) 777, 961, 974
AlwNI CAGNNNCTG 1 cut(s) 110
AoxI GGCC 8 cut(s) 158, 175, 399, 519, 667, 826, 939, 1041
ApeKI GCWGC 3 cut(s) 55, 284, 1427
ApoI RAATTY 1 cut(s) 847
ArsI GACNNNNNNTTYG 2 cut(s) 806, 838
AseI ATTAAT 1 cut(s) 1265
AspLEI GCGC 2 cut(s) 702, 1432
AspS9I GGNCC 5 cut(s) 176, 667, 875, 939, 946
AsuC2I CCSGG 3 cut(s) 954, 1009, 1319
AsuHPI GGTGA 4 cut(s) 733, 755, 1106, 1392
AvaII GGWCC 2 cut(s) 875, 946
BalI TGGCCA 1 cut(s) 828
BamHI GGATCC 1 cut(s) 966
BanI GGYRCC 2 cut(s) 4, 635
BauI CACGAG 1 cut(s) 465
BbvI GCAGC 3 cut(s) 42, 296, 1414
BccI CCATC 6 cut(s) 444, 770, 876, 1097, 1112, 1142
BceAI ACGGC 3 cut(s) 414, 878, 1006
BciT130I CCWGG 1 cut(s) 375
BcnI CCSGG 3 cut(s) 954, 1009, 1319
BcoDI GTCTC 2 cut(s) 924, 1245
BfaI CTAG 3 cut(s) 923, 1208, 1311
BfoI RGCGCY 1 cut(s) 703
BfuAI ACCTGC 1 cut(s) 623
BglI GCCNNNNNGGC 1 cut(s) 1049
BisI GCNGC 9 cut(s) 56, 241, 285, 891, 1053, 1060, 1428, 1433, 1436
BlpI GCTNAGC 1 cut(s) 906
BlsI GCNGC 9 cut(s) 57, 242, 286, 892, 1054, 1061, 1429, 1434, 1437
Bme1390I CCNGG 4 cut(s) 375, 954, 1009, 1319
Bme18I GGWCC 2 cut(s) 875, 946
BmgT120I GGNCC 5 cut(s) 176, 667, 875, 939, 946
BmiI GGNNCC 7 cut(s) 6, 217, 552, 637, 677, 877, 968
BmrFI CCNGG 4 cut(s) 375, 954, 1009, 1319
BoxI GACNNNNGTC 1 cut(s) 382
BpmI CTGGAG 1 cut(s) 1264
Bpu1102I GCTNAGC 1 cut(s) 906
BpuMI CCSGG 3 cut(s) 954, 1009, 1319
Bsa29I ATCGAT 1 cut(s) 1389
BsaBI GATNNNNATC 1 cut(s) 965
BsaI GGTCTC 1 cut(s) 924
BsaJI CCNNGG 6 cut(s) 161, 179, 838, 942, 952, 1318
BsaWI WCCGGW 1 cut(s) 218
BsaXI ACNNNNNCTCC 2 cut(s) 849, 879
Bsc4I CCNNNNNNNGG 5 cut(s) 180, 616, 736, 924, 976
Bse118I RCCGGY 2 cut(s) 173, 521
Bse1I ACTGG 2 cut(s) 8, 821
Bse3DI GCAATG 1 cut(s) 301
Bse8I GATNNNNATC 1 cut(s) 965
BseBI CCWGG 1 cut(s) 375
BseCI ATCGAT 1 cut(s) 1389
BseDI CCNNGG 6 cut(s) 161, 179, 838, 942, 952, 1318
BseGI GGATG 3 cut(s) 455, 1036, 1123
BseJI GATNNNNATC 1 cut(s) 965
BseLI CCNNNNNNNGG 5 cut(s) 180, 616, 736, 924, 976
BseMI GCAATG 1 cut(s) 301
BseMII CTCAG 2 cut(s) 374, 920
BseNI ACTGG 2 cut(s) 8, 821
BseRI GAGGAG 2 cut(s) 874, 1233
BseXI GCAGC 3 cut(s) 42, 296, 1414
BseYI CCCAGC 2 cut(s) 1022, 1062
BshFI GGCC 8 cut(s) 160, 177, 401, 521, 669, 828, 941, 1043
BshNI GGYRCC 2 cut(s) 4, 635
BshVI ATCGAT 1 cut(s) 1389
BsiSI CCGG 7 cut(s) 174, 219, 522, 639, 954, 1008, 1318
BslFI GGGAC 2 cut(s) 1085, 1362
BslI CCNNNNNNNGG 5 cut(s) 180, 616, 736, 924, 976
BsmAI GTCTC 2 cut(s) 924, 1245
BsmFI GGGAC 2 cut(s) 1085, 1362
BsmI GAATGC 1 cut(s) 1226
BsnI GGCC 8 cut(s) 160, 177, 401, 521, 669, 828, 941, 1043
Bso31I GGTCTC 1 cut(s) 924
Bsp143I GATC 3 cut(s) 782, 966, 1386
Bsp1720I GCTNAGC 1 cut(s) 906
Bsp19I CCATGG 1 cut(s) 942
BspANI GGCC 8 cut(s) 160, 177, 401, 521, 669, 828, 941, 1043
BspCNI CTCAG 2 cut(s) 373, 919
BspDI ATCGAT 1 cut(s) 1389
BspHI TCATGA 1 cut(s) 124
BspLI GGNNCC 7 cut(s) 6, 217, 552, 637, 677, 877, 968
BspMI ACCTGC 1 cut(s) 623
BspPI GGATC 3 cut(s) 777, 961, 974
BspT107I GGYRCC 2 cut(s) 4, 635
BspTNI GGTCTC 1 cut(s) 924
BsrBI CCGCTC 1 cut(s) 1059
BsrDI GCAATG 1 cut(s) 301
BsrFI RCCGGY 2 cut(s) 173, 521
BsrI ACTGG 2 cut(s) 8, 821
BssAI RCCGGY 2 cut(s) 173, 521
BssECI CCNNGG 6 cut(s) 161, 179, 838, 942, 952, 1318
BssMI GATC 3 cut(s) 782, 966, 1386
BssSI CACGAG 1 cut(s) 465
BssT1I CCWWGG 4 cut(s) 161, 179, 838, 942
Bst2BI CACGAG 1 cut(s) 465
Bst2UI CCWGG 1 cut(s) 375
Bst6I CTCTTC 1 cut(s) 1251
BstC8I GCNNGC 5 cut(s) 189, 238, 318, 523, 1057
BstDEI CTNAG 2 cut(s) 360, 906
BstDSI CCRYGG 1 cut(s) 942
BstF5I GGATG 3 cut(s) 455, 1036, 1123
BstH2I RGCGCY 1 cut(s) 703
BstHHI GCGC 2 cut(s) 702, 1432
BstKTI GATC 3 cut(s) 785, 969, 1389
BstMAI GTCTC 2 cut(s) 924, 1245
BstMBI GATC 3 cut(s) 782, 966, 1386
BstMWI GCNNNNNNNGC 3 cut(s) 887, 1009, 1049
BstNI CCWGG 1 cut(s) 375
BstNSI RCATGY 2 cut(s) 191, 1240
BstPAI GACNNNNGTC 1 cut(s) 382
BstSCI CCNGG 4 cut(s) 373, 952, 1007, 1317
BstV1I GCAGC 3 cut(s) 42, 296, 1414
BstX2I RGATCY 2 cut(s) 782, 966
BstYI RGATCY 2 cut(s) 782, 966
Bsu15I ATCGAT 1 cut(s) 1389
BsuRI GGCC 8 cut(s) 160, 177, 401, 521, 669, 828, 941, 1043
BsuTUI ATCGAT 1 cut(s) 1389
BtgI CCRYGG 1 cut(s) 942
BtsCI GGATG 3 cut(s) 455, 1036, 1123
BtsIMutI CAGTG 1 cut(s) 368
BveI ACCTGC 1 cut(s) 623
Cac8I GCNNGC 5 cut(s) 189, 238, 318, 523, 1057
CaiI CAGNNNCTG 1 cut(s) 110
CciI TCATGA 1 cut(s) 124
CfoI GCGC 2 cut(s) 702, 1432
Cfr10I RCCGGY 2 cut(s) 173, 521
Cfr13I GGNCC 5 cut(s) 176, 667, 875, 939, 946
ClaI ATCGAT 1 cut(s) 1389
Csp6I GTAC 3 cut(s) 222, 514, 541
CviQI GTAC 3 cut(s) 222, 514, 541
DdeI CTNAG 2 cut(s) 360, 906
DpnI GATC 3 cut(s) 784, 968, 1388
DpnII GATC 3 cut(s) 782, 966, 1386
DraIII CACNNNGTG 1 cut(s) 467
EaeI YGGCCR 2 cut(s) 399, 826
Eam1104I CTCTTC 1 cut(s) 1251
EarI CTCTTC 1 cut(s) 1251
EciI GGCGGA 1 cut(s) 877
Eco130I CCWWGG 4 cut(s) 161, 179, 838, 942
Eco31I GGTCTC 1 cut(s) 924
Eco47I GGWCC 2 cut(s) 875, 946
Eco57I CTGAAG 1 cut(s) 716
EcoRII CCWGG 1 cut(s) 373
EcoT14I CCWWGG 4 cut(s) 161, 179, 838, 942
EcoT22I ATGCAT 1 cut(s) 301
ErhI CCWWGG 4 cut(s) 161, 179, 838, 942
FaqI GGGAC 2 cut(s) 1085, 1362
FauI CCCGC 1 cut(s) 886
FblI GTMKAC 1 cut(s) 1289
Fnu4HI GCNGC 9 cut(s) 56, 241, 285, 891, 1053, 1060, 1428, 1433, 1436
FokI GGATG 3 cut(s) 462, 1043, 1130
Fsp4HI GCNGC 9 cut(s) 56, 241, 285, 891, 1053, 1060, 1428, 1433, 1436
FspBI CTAG 3 cut(s) 923, 1208, 1311
GlaI GCGC 2 cut(s) 701, 1431
GluI GCNGC 9 cut(s) 56, 241, 285, 891, 1053, 1060, 1428, 1433, 1436
GsaI CCCAGC 2 cut(s) 1026, 1066
GsuI CTGGAG 1 cut(s) 1264
HaeII RGCGCY 1 cut(s) 703
HaeIII GGCC 8 cut(s) 160, 177, 401, 521, 669, 828, 941, 1043
HapII CCGG 7 cut(s) 174, 219, 522, 639, 954, 1008, 1318
HhaI GCGC 2 cut(s) 702, 1432
Hin6I GCGC 2 cut(s) 700, 1430
HinP1I GCGC 2 cut(s) 700, 1430
HincII GTYRAC 1 cut(s) 988
HindII GTYRAC 1 cut(s) 988
HindIII AAGCTT 1 cut(s) 131
HinfI GANTC 5 cut(s) 595, 858, 1204, 1333, 1405
HpaII CCGG 7 cut(s) 174, 219, 522, 639, 954, 1008, 1318
HphI GGTGA 4 cut(s) 733, 755, 1106, 1392
Hpy166II GTNNAC 5 cut(s) 116, 582, 875, 988, 1290
Hpy188I TCNGA 4 cut(s) 714, 761, 971, 1203
Hpy188III TCNNGA 4 cut(s) 125, 467, 622, 1208
Hpy8I GTNNAC 5 cut(s) 116, 582, 875, 988, 1290
Hpy99I CGWCG 4 cut(s) 632, 917, 1079, 1082
HpyAV CCTTC 9 cut(s) 203, 257, 501, 766, 844, 974, 1085, 1118, 1228
HpyCH4IV ACGT 3 cut(s) 90, 627, 1080
HpyCH4V TGCA 6 cut(s) 74, 80, 299, 369, 456, 1226
HpyF10VI GCNNNNNNNGC 3 cut(s) 887, 1009, 1049
HpyF3I CTNAG 2 cut(s) 360, 906
HpySE526I ACGT 3 cut(s) 90, 627, 1080
HspAI GCGC 2 cut(s) 700, 1430
KroI GCCGGC 1 cut(s) 521
KroNI GCCGGC 1 cut(s) 523
Kzo9I GATC 3 cut(s) 782, 966, 1386
LmnI GCTCC 4 cut(s) 155, 221, 233, 1132
Lsp1109I GCAGC 3 cut(s) 42, 296, 1414
MaeI CTAG 3 cut(s) 923, 1208, 1311
MaeII ACGT 3 cut(s) 90, 627, 1080
MaeIII GTNAC 4 cut(s) 384, 394, 646, 721
MalI GATC 3 cut(s) 784, 968, 1388
MbiI CCGCTC 1 cut(s) 1059
MboI GATC 3 cut(s) 782, 966, 1386
MboII GAAGA 5 cut(s) 38, 260, 792, 1223, 1238
MfeI CAATTG 1 cut(s) 409
MflI RGATCY 2 cut(s) 782, 966
MlsI TGGCCA 1 cut(s) 828
MluCI AATT 4 cut(s) 409, 504, 847, 1194
MluNI TGGCCA 1 cut(s) 828
MlyI GAGTC 2 cut(s) 852, 1399
MmeI TCCRAC 1 cut(s) 255
Mox20I TGGCCA 1 cut(s) 828
Mph1103I ATGCAT 1 cut(s) 301
MroNI GCCGGC 1 cut(s) 521
MscI TGGCCA 1 cut(s) 828
MseI TTAA 4 cut(s) 258, 897, 1137, 1265
MslI CAYNNNNRTG 1 cut(s) 75
Msp20I TGGCCA 1 cut(s) 828
MspA1I CMGCKG 3 cut(s) 437, 694, 1284
MspI CCGG 7 cut(s) 174, 219, 522, 639, 954, 1008, 1318
MspR9I CCNGG 4 cut(s) 375, 954, 1009, 1319
MunI CAATTG 1 cut(s) 409
Mva1269I GAATGC 1 cut(s) 1226
MvaI CCWGG 1 cut(s) 375
MwoI GCNNNNNNNGC 3 cut(s) 887, 1009, 1049
NaeI GCCGGC 1 cut(s) 523
NciI CCSGG 3 cut(s) 954, 1009, 1319
NcoI CCATGG 1 cut(s) 942
NdeII GATC 3 cut(s) 782, 966, 1386
NgoMIV GCCGGC 1 cut(s) 521
NlaIV GGNNCC 7 cut(s) 6, 217, 552, 637, 677, 877, 968
NmeAIII GCCGAG 1 cut(s) 535
NmuCI GTSAC 3 cut(s) 384, 646, 721
NsiI ATGCAT 1 cut(s) 301
NspI RCATGY 2 cut(s) 191, 1240
PaeI GCATGC 1 cut(s) 191
PagI TCATGA 1 cut(s) 124
PaqCI CACCTGC 1 cut(s) 623
PciI ACATGT 1 cut(s) 1236
PctI GAATGC 1 cut(s) 1226
PdiI GCCGGC 1 cut(s) 523
PfeI GAWTC 3 cut(s) 595, 1204, 1333
PflFI GACNNNGTC 1 cut(s) 1353
PflMI CCANNNNNTGG 2 cut(s) 616, 736
PkrI GCNGC 9 cut(s) 57, 242, 286, 892, 1054, 1061, 1429, 1434, 1437
PleI GAGTC 2 cut(s) 852, 1399
PpsI GAGTC 2 cut(s) 852, 1399
PscI ACATGT 1 cut(s) 1236
PshAI GACNNNNGTC 1 cut(s) 382
PshBI ATTAAT 1 cut(s) 1265
Psp1406I AACGTT 1 cut(s) 90
Psp6I CCWGG 1 cut(s) 373
PspFI CCCAGC 2 cut(s) 1022, 1062
PspGI CCWGG 1 cut(s) 373
PspN4I GGNNCC 7 cut(s) 6, 217, 552, 637, 677, 877, 968
PspPI GGNCC 5 cut(s) 176, 667, 875, 939, 946
PstNI CAGNNNCTG 1 cut(s) 110
PsuI RGATCY 2 cut(s) 782, 966
PsyI GACNNNGTC 1 cut(s) 1353
PvuII CAGCTG 1 cut(s) 1284
RsaI GTAC 3 cut(s) 223, 515, 542
RsaNI GTAC 3 cut(s) 222, 514, 541
RseI CAYNNNNRTG 1 cut(s) 75
SaqAI TTAA 4 cut(s) 258, 897, 1137, 1265
SatI GCNGC 9 cut(s) 56, 241, 285, 891, 1053, 1060, 1428, 1433, 1436
Sau3AI GATC 3 cut(s) 782, 966, 1386
Sau96I GGNCC 5 cut(s) 176, 667, 875, 939, 946
SchI GAGTC 2 cut(s) 852, 1399
ScrFI CCNGG 4 cut(s) 375, 954, 1009, 1319
SinI GGWCC 2 cut(s) 875, 946
SmiMI CAYNNNNRTG 1 cut(s) 75
SphI GCATGC 1 cut(s) 191
Sse9I AATT 4 cut(s) 409, 504, 847, 1194
SspMI CTAG 3 cut(s) 923, 1208, 1311
StyD4I CCNGG 4 cut(s) 373, 952, 1007, 1317
StyI CCWWGG 4 cut(s) 161, 179, 838, 942
TaiI ACGT 3 cut(s) 93, 630, 1083
TaqI TCGA 3 cut(s) 477, 856, 1389
TasI AATT 4 cut(s) 409, 504, 847, 1194
TatI WGTACW 1 cut(s) 513
TauI GCSGC 6 cut(s) 243, 893, 1055, 1062, 1435, 1438
TfiI GAWTC 3 cut(s) 595, 1204, 1333
Tru1I TTAA 4 cut(s) 258, 897, 1137, 1265
Tru9I TTAA 4 cut(s) 258, 897, 1137, 1265
TscAI CASTG 1 cut(s) 368
TseFI GTSAC 3 cut(s) 384, 646, 721
TseI GCWGC 3 cut(s) 55, 284, 1427
Tsp45I GTSAC 3 cut(s) 384, 646, 721
TspDTI ATGAA 7 cut(s) 21, 59, 125, 318, 351, 483, 1047
TspGWI ACGGA 2 cut(s) 97, 1155
TspRI CASTG 1 cut(s) 368
Tth111I GACNNNGTC 1 cut(s) 1353
Van91I CCANNNNNTGG 2 cut(s) 616, 736
VpaK11BI GGWCC 2 cut(s) 875, 946
VspI ATTAAT 1 cut(s) 1265
XapI RAATTY 1 cut(s) 847
XbaI TCTAGA 1 cut(s) 1207
XceI RCATGY 2 cut(s) 191, 1240
XcmI CCANNNNNNNNNTGG 1 cut(s) 737
XmiI GTMKAC 1 cut(s) 1289
XspI CTAG 3 cut(s) 923, 1208, 1311
Zsp2I ATGCAT 1 cut(s) 301
Using CommOnly database (standard laboratory enzymes). Scanned on CDS sequence.