Rmu_sc0007349.1_g000001

amidase C869.01

Design Primers CRISPR Design

Basic Information

Type: gene
Biological Identity
rosa_multiflora
Rmu_sc0007349.1
OG0000427
Physical Location & Seq
Forward (+)
9263 .. 11393
2131 bp
Loading structure...
UTR
Exon/CDS
Intron
Rmu_sc0007349.1_g000001.1.cds

Sequence Viewer

Length: 1575 bp
atggccaaaactccgttgatcagctcaaattcttgggttgttggtcactggcagtctggcactccattactgccattatcatccaaggctaactatcttctcctctcgtttttgcttctaatttgcattctatcatatggctcacaaaaactcacggccaaaaggcccaaaatcacaatcgaagaagctactgttgatgatctccagcttgctttcaagctcaacactctgacctcgaggcaacttgttctgttctacatggatgagattcaaaaactgaacaaaaaccttagggctgtgatagaggtgaaccctgatgctttatcccaagctgatatagctgacaaacaacgcagagaatgcaaccacacactgcctaagcttcatggcattcctattctgctcaaggataccatagcaaccaaggacaaactcaacaccacagctggatcgtttgccttgttggggtccattgttcctcgagatgcaggtgttgtttctaagctaaggaggtctggagctatcattttagggaaggctagcttgagtgaatggtgccatttcaggtcattcatagcaccttctggatggagcgccagaggttcccaagggatgaatccttacaattcatcggtggaagtttgtggatcaagtagtggacctgcaatatcagtagctgctaatatggtttctgtgacattagggacggagactgacagctccattatctgtccggctagctttaactctgtggtgggcatcaaacccactgttggtctcactagtcgagcaggagtcataccaatcagtccaagacaggacacagttgggccgatttgtaggactgtagcggatgctgttcacgtccttgatatcattgttggcattgataggaatgataatgcaacaagtgaagcatcaaagtacatcccagtgggaggctatggacagtttctcaagcataatggtctcgaaggaaagagattggggatagtaaatatgttatttgattcaatcaaggacgatgcttttctgaaacaaacttttgtgcagcattttaccacactgaggaaacaaggtgcaattttggttgacaatcttgaaatagccaatattgaagatgttctggattctcattcaagtggagaatttgatgctatattagctgaattcaaaatagccttgaatgagtacctaagtgacctagtgaaatctccagtgaagtccttaagggaggtgatagagttcaacaagaatcattcaaaggtggtgagtgtatctggtcacagacatggctcacaaaaactcacggccaaaaggcccaaaatcacaatcgaagaagctactgttgatgatctccagcttgctttcaagctcaacactctgactctggattctcattcaagtggagaatttgatgctatattagctgaattcaaaatagccttgaatgagtacctaagtgacctagtaaaatctccagtgaagtccttaagggaggtgattgagttcaacaagaatcattcaaaggtggtgagtgtatctggtcacagacctctttgctcatgcttttaa
Functional Annotation
Gene Ontology
Molecular Function
Biological Process
Cellular Component
GO:0003674 GO:0003824 GO:0016787 GO:0016810 GO:0016811
KEGG Pathways
Metabolic & Signaling
ko00330 ko00360 ko00380 ko00627 ko00643 ko01120 map00330 map00360 map00380 map00627 map00643 map01120
Pfam Domains
Protein Families
Amidase

Protein Analysis

524

Amino Acids

57.03

Weight (kDa)

8.61

Isoelectric Point (pI)

41.28

Instability Index
Protein Domains (Pfam)
No Pfam domains detected for this protein.
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: OG0000427)

Species Orthologous Gene IDs
arabidopsis_thaliana AT4G34880 AT4G34880
fragaria_vesca FvH4_2g01710 FvH4_2g01720 FvH4_2g01722
malus_domestica MD05G1117000.v1.1 MD05G1117200.v1.1 MD10G1120200.v1.1 MD10G1120300.v1.1 MD10G1120400.v1.1
prunus_persica Prupe.8G161600_v2.0.a1 Prupe.8G161700_v2.0.a1 Prupe.8G161700_v2.0.a1 Prupe.8G161800_v2.0.a1 Prupe.8G161900_v2.0.a1
pyrus_communis pycom05g06760 pycom10g10360 pycom10g10370 pycom17g11700
rosa_chinensis RchiOBHm_Chr6g0244611 RchiOBHm_Chr6g0244621 RchiOBHm_Chr6g0244631 RchiOBHm_Chr6g0244671 RchiOBHm_Chr6g0244701 RchiOBHm_Chr6g0244721 RchiOBHm_Chr6g0244741 RchiOBHm_Chr6g0244751
rosa_laevigata RLG00000015331 RLG00000015332 RLG00000015333 RLG00000015334 RLG00000015336 RLG00000015338 RLG00000015339 RLG00000015340
rosa_multiflora Rmu_co8154324.1_g000001 Rmu_co8259027.1_g000001 Rmu_co8319583.1_g000001 Rmu_sc0002064.1_g000004 Rmu_sc0002522.1_g000021 Rmu_sc0002522.1_g000022 Rmu_sc0002522.1_g000023 Rmu_sc0002522.1_g000026 Rmu_sc0002522.1_g000027 Rmu_sc0006257.1_g000002 Rmu_sc0006635.1_g000001 Rmu_sc0006635.1_g000002 Rmu_sc0007349.1_g000001 Rmu_sc0011292.1_g000009 Rmu_sc0011292.1_g000010 Rmu_sc0039466.1_g000001
rosa_roxburghii Rroxscaffold_7G00215530 Rroxscaffold_7G00215550 Rroxscaffold_7G00215560 Rroxscaffold_7G00215570 Rroxscaffold_7G00215610 Rroxscaffold_7G00215620 Rroxscaffold_7G00215630 Rroxscaffold_7G00215640
rosa_rugosa Rorug05G0509900 Rorug05G0510000 Rorug05G0510000 Rorug05G0510100 Rorug05G0510100 Rorug05G0510200 Rorug05G0510300 Rorug05G0510400
rosa_samantha Rh6BG020500 Rh6BG020600 Rh6BG020700 Rh6BG020900 Rh6BG021000 Rh6BG021100 Rh6BG021200 Rh6CG018800 Rh6CG019000 Rh6CG019100 Rh6CG019200 Rh6CG019300 Rh6CG019400 Rh6CG019600 Rh6CG019800 Rh6DG019900 Rh6DG020100 Rh6DG020200 Rh6DG020400
rosa_wichuraiana Rw0G014670 Rw6G000010 Rw6G000020 Rw6G002100 Rw6G002110 Rw6G002120 Rw6G002140 Rw6G002150 Rw6G002160 Rw6G005110

Restriction Enzyme Sites

1 / 10
Enzyme Recognition Site Cut Count Positions (bp)
AarI CACCTGC 1 cut(s) 479
AbsI CCTCGAGG 1 cut(s) 235
Acc36I ACCTGC 2 cut(s) 479, 670
AccB1I GGYRCC 1 cut(s) 555
AciI CCGC 1 cut(s) 851
AclWI GGATC 2 cut(s) 457, 655
AcoI YGGCCR 3 cut(s) 3, 156, 1311
AcsI RAATTY 5 cut(s) 28, 1148, 1169, 1412, 1433
AfaI GTAC 3 cut(s) 926, 1193, 1457
AfiI CCNNNNNNNGG 4 cut(s) 465, 773, 938, 1068
AflII CTTAAG 2 cut(s) 1228, 1492
AhlI ACTAGT 1 cut(s) 782
AjiI CACGTC 1 cut(s) 865
Alw26I GTCTC 3 cut(s) 704, 782, 974
AlwI GGATC 2 cut(s) 457, 655
AlwNI CAGNNNCTG 1 cut(s) 677
Ama87I CYCGRG 2 cut(s) 235, 480
AoxI GGCC 6 cut(s) 3, 156, 164, 830, 1311, 1319
ApeKI GCWGC 2 cut(s) 677, 1051
ApoI RAATTY 5 cut(s) 28, 1148, 1169, 1412, 1433
Asp700I GAANNNNTTC 1 cut(s) 1122
AspLEI GCGC 1 cut(s) 596
AspS9I GGNCC 5 cut(s) 165, 468, 659, 830, 1320
AsuHPI GGTGA 5 cut(s) 319, 1249, 1282, 1513, 1546
AsuNHI GCTAGC 2 cut(s) 539, 737
AvaI CYCGRG 2 cut(s) 235, 480
AvaII GGWCC 2 cut(s) 468, 659
AxyI CCTNAGG 1 cut(s) 290
BalI TGGCCA 1 cut(s) 5
BanI GGYRCC 1 cut(s) 555
BbvI GCAGC 2 cut(s) 664, 1063
BccI CCATC 1 cut(s) 582
BceAI ACGGC 2 cut(s) 171, 1326
BciVI GTATCC 1 cut(s) 403
BclI TGATCA 1 cut(s) 18
BcoDI GTCTC 3 cut(s) 704, 782, 974
BcuI ACTAGT 1 cut(s) 782
BfaI CTAG 5 cut(s) 540, 738, 783, 1205, 1469
BfmI CTRYAG 1 cut(s) 846
BfoI RGCGCY 1 cut(s) 597
BfrI CTTAAG 2 cut(s) 1228, 1492
BfuAI ACCTGC 2 cut(s) 479, 670
BfuI GTATCC 1 cut(s) 403
BisI GCNGC 2 cut(s) 678, 1052
BlsI GCNGC 2 cut(s) 679, 1053
Bme18I GGWCC 2 cut(s) 468, 659
BmeT110I CYCGRG 2 cut(s) 235, 480
BmgBI CACGTC 1 cut(s) 865
BmgT120I GGNCC 5 cut(s) 165, 468, 659, 830, 1320
BmiI GGNNCC 3 cut(s) 469, 557, 604
BmrI ACTGGG 1 cut(s) 926
BmsI GCATC 8 cut(s) 307, 475, 768, 844, 926, 1015, 1144, 1408
BmtI GCTAGC 2 cut(s) 543, 741
BmuI ACTGGG 1 cut(s) 926
BpmI CTGGAG 5 cut(s) 188, 537, 1200, 1343, 1464
Bpu10I CCTNAGC 2 cut(s) 378, 506
BpuEI CTTGAG 3 cut(s) 389, 565, 941
BsaI GGTCTC 2 cut(s) 782, 974
BsaJI CCNNGG 3 cut(s) 84, 423, 607
Bsc4I CCNNNNNNNGG 4 cut(s) 465, 773, 938, 1068
Bse1I ACTGG 4 cut(s) 53, 932, 1217, 1481
Bse21I CCTNAGG 1 cut(s) 290
BseDI CCNNGG 3 cut(s) 84, 423, 607
BseGI GGATG 6 cut(s) 80, 268, 593, 618, 859, 927
BseLI CCNNNNNNNGG 4 cut(s) 465, 773, 938, 1068
BseMII CTCAG 1 cut(s) 1058
BseNI ACTGG 4 cut(s) 53, 932, 1217, 1481
BseRI GAGGAG 1 cut(s) 92
BseXI GCAGC 2 cut(s) 664, 1063
BsgI GTGCAG 1 cut(s) 1070
BshFI GGCC 6 cut(s) 5, 158, 166, 832, 1313, 1321
BshNI GGYRCC 1 cut(s) 555
BsiHKCI CYCGRG 2 cut(s) 235, 480
BsiSI CCGG 1 cut(s) 734
BslFI GGGAC 1 cut(s) 718
BslI CCNNNNNNNGG 4 cut(s) 465, 773, 938, 1068
BsmAI GTCTC 3 cut(s) 704, 782, 974
BsmFI GGGAC 1 cut(s) 718
BsmI GAATGC 3 cut(s) 126, 365, 390
BsnI GGCC 6 cut(s) 5, 158, 166, 832, 1313, 1321
Bso31I GGTCTC 2 cut(s) 782, 974
BsoBI CYCGRG 2 cut(s) 235, 480
Bsp143I GATC 5 cut(s) 18, 199, 449, 647, 1354
BspACI CCGC 1 cut(s) 851
BspANI GGCC 6 cut(s) 5, 158, 166, 832, 1313, 1321
BspCNI CTCAG 1 cut(s) 1059
BspLI GGNNCC 3 cut(s) 469, 557, 604
BspMI ACCTGC 2 cut(s) 479, 670
BspOI GCTAGC 2 cut(s) 543, 741
BspPI GGATC 2 cut(s) 457, 655
BspT107I GGYRCC 1 cut(s) 555
BspTI CTTAAG 2 cut(s) 1228, 1492
BspTNI GGTCTC 2 cut(s) 782, 974
BsrI ACTGG 4 cut(s) 53, 932, 1217, 1481
BssECI CCNNGG 3 cut(s) 84, 423, 607
BssMI GATC 5 cut(s) 18, 199, 449, 647, 1354
BssT1I CCWWGG 3 cut(s) 84, 423, 607
Bst4CI ACNGT 6 cut(s) 193, 772, 826, 847, 951, 1348
BstAFI CTTAAG 2 cut(s) 1228, 1492
BstAPI GCANNNNNTGC 1 cut(s) 360
BstC8I GCNNGC 4 cut(s) 210, 541, 739, 1365
BstDEI CTNAG 7 cut(s) 290, 378, 501, 506, 1067, 1196, 1460
BstF5I GGATG 6 cut(s) 80, 268, 593, 618, 859, 927
BstH2I RGCGCY 1 cut(s) 597
BstHHI GCGC 1 cut(s) 596
BstKTI GATC 5 cut(s) 21, 202, 452, 650, 1357
BstMAI GTCTC 3 cut(s) 704, 782, 974
BstMBI GATC 5 cut(s) 18, 199, 449, 647, 1354
BstMWI GCNNNNNNNGC 4 cut(s) 338, 360, 1163, 1427
BstSFI CTRYAG 1 cut(s) 846
BstV1I GCAGC 2 cut(s) 664, 1063
Bsu36I CCTNAGG 1 cut(s) 290
BsuI GTATCC 1 cut(s) 403
BsuRI GGCC 6 cut(s) 5, 158, 166, 832, 1313, 1321
BtrI CACGTC 1 cut(s) 865
BtsCI GGATG 6 cut(s) 80, 268, 593, 618, 859, 927
BtsI GCAGTG 1 cut(s) 371
BtsIMutI CAGTG 7 cut(s) 46, 371, 768, 939, 1064, 1224, 1488
BveI ACCTGC 2 cut(s) 479, 670
Cac8I GCNNGC 4 cut(s) 210, 541, 739, 1365
CaiI CAGNNNCTG 1 cut(s) 677
CfoI GCGC 1 cut(s) 596
Cfr13I GGNCC 5 cut(s) 165, 468, 659, 830, 1320
Csp6I GTAC 3 cut(s) 925, 1192, 1456
CviAII CATG 4 cut(s) 259, 386, 1292, 1566
CviQI GTAC 3 cut(s) 925, 1192, 1456
DdeI CTNAG 7 cut(s) 290, 378, 501, 506, 1067, 1196, 1460
DpnI GATC 5 cut(s) 20, 201, 451, 649, 1356
DpnII GATC 5 cut(s) 18, 199, 449, 647, 1354
EaeI YGGCCR 3 cut(s) 3, 156, 1311
Eco130I CCWWGG 3 cut(s) 84, 423, 607
Eco31I GGTCTC 2 cut(s) 782, 974
Eco32I GATATC 1 cut(s) 874
Eco47I GGWCC 2 cut(s) 468, 659
Eco81I CCTNAGG 1 cut(s) 290
Eco88I CYCGRG 2 cut(s) 235, 480
EcoRI GAATTC 2 cut(s) 1169, 1433
EcoRV GATATC 1 cut(s) 874
EcoT14I CCWWGG 3 cut(s) 84, 423, 607
ErhI CCWWGG 3 cut(s) 84, 423, 607
FaeI CATG 4 cut(s) 262, 389, 1295, 1569
FalI AAGNNNNNCTT 2 cut(s) 527, 559
FaqI GGGAC 1 cut(s) 718
FatI CATG 4 cut(s) 258, 385, 1291, 1565
FauNDI CATATG 1 cut(s) 136
FbaI TGATCA 1 cut(s) 18
Fnu4HI GCNGC 2 cut(s) 678, 1052
FokI GGATG 6 cut(s) 67, 275, 600, 625, 866, 914
Fsp4HI GCNGC 2 cut(s) 678, 1052
FspBI CTAG 5 cut(s) 540, 738, 783, 1205, 1469
GlaI GCGC 1 cut(s) 595
GluI GCNGC 2 cut(s) 678, 1052
GsuI CTGGAG 5 cut(s) 188, 537, 1200, 1343, 1464
HaeII RGCGCY 1 cut(s) 597
HaeIII GGCC 6 cut(s) 5, 158, 166, 832, 1313, 1321
HapII CCGG 1 cut(s) 734
HhaI GCGC 1 cut(s) 596
Hin1II CATG 4 cut(s) 262, 389, 1295, 1569
Hin6I GCGC 1 cut(s) 594
HinP1I GCGC 1 cut(s) 594
HincII GTYRAC 1 cut(s) 1093
HindII GTYRAC 1 cut(s) 1093
HindIII AAGCTT 1 cut(s) 380
HinfI GANTC 9 cut(s) 268, 616, 795, 1010, 1130, 1255, 1387, 1394, 1519
HpaII CCGG 1 cut(s) 734
HphI GGTGA 5 cut(s) 319, 1249, 1282, 1513, 1546
Hpy166II GTNNAC 4 cut(s) 310, 659, 862, 1093
Hpy188I TCNGA 3 cut(s) 231, 1035, 1386
Hpy188III TCNNGA 7 cut(s) 482, 516, 585, 971, 1100, 1127, 1391
Hpy8I GTNNAC 4 cut(s) 310, 659, 862, 1093
HpyAV CCTTC 3 cut(s) 529, 591, 968
HpyCH4III ACNGT 6 cut(s) 193, 772, 826, 847, 951, 1348
HpyCH4IV ACGT 1 cut(s) 864
HpyCH4V TGCA 7 cut(s) 126, 363, 488, 665, 905, 1051, 1082
HpyF10VI GCNNNNNNNGC 4 cut(s) 338, 360, 1163, 1427
HpyF3I CTNAG 7 cut(s) 290, 378, 501, 506, 1067, 1196, 1460
HpySE526I ACGT 1 cut(s) 864
Hsp92II CATG 4 cut(s) 262, 389, 1295, 1569
HspAI GCGC 1 cut(s) 594
Ksp22I TGATCA 1 cut(s) 18
Kzo9I GATC 5 cut(s) 18, 199, 449, 647, 1354
LmnI GCTCC 3 cut(s) 518, 591, 725
Lsp1109I GCAGC 2 cut(s) 664, 1063
LweI GCATC 8 cut(s) 307, 475, 768, 844, 926, 1015, 1144, 1408
MaeI CTAG 5 cut(s) 540, 738, 783, 1205, 1469
MaeII ACGT 1 cut(s) 864
MaeIII GTNAC 6 cut(s) 44, 694, 1199, 1283, 1463, 1547
MalI GATC 5 cut(s) 20, 201, 451, 649, 1356
MboI GATC 5 cut(s) 18, 199, 449, 647, 1354
MboII GAAGA 4 cut(s) 89, 194, 1130, 1349
MlsI TGGCCA 1 cut(s) 5
MluCI AATT 8 cut(s) 28, 120, 625, 1083, 1148, 1169, 1412, 1433
MluNI TGGCCA 1 cut(s) 5
MlyI GAGTC 2 cut(s) 804, 1381
Mox20I TGGCCA 1 cut(s) 5
MroXI GAANNNNTTC 1 cut(s) 1122
MscI TGGCCA 1 cut(s) 5
MseI TTAA 4 cut(s) 744, 1229, 1493, 1573
MslI CAYNNNNRTG 4 cut(s) 932, 1140, 1290, 1404
Msp20I TGGCCA 1 cut(s) 5
MspA1I CMGCKG 1 cut(s) 446
MspCI CTTAAG 2 cut(s) 1228, 1492
MspI CCGG 1 cut(s) 734
Mva1269I GAATGC 3 cut(s) 126, 365, 390
MwoI GCNNNNNNNGC 4 cut(s) 338, 360, 1163, 1427
NdeI CATATG 1 cut(s) 136
NdeII GATC 5 cut(s) 18, 199, 449, 647, 1354
NheI GCTAGC 2 cut(s) 539, 737
NlaIII CATG 4 cut(s) 262, 389, 1295, 1569
NlaIV GGNNCC 3 cut(s) 469, 557, 604
NmuCI GTSAC 6 cut(s) 44, 694, 1199, 1283, 1463, 1547
PaeR7I CTCGAG 2 cut(s) 235, 480
PaqCI CACCTGC 1 cut(s) 479
PctI GAATGC 3 cut(s) 126, 365, 390
PdmI GAANNNNTTC 1 cut(s) 1122
PfeI GAWTC 7 cut(s) 268, 616, 1010, 1130, 1255, 1394, 1519
PkrI GCNGC 2 cut(s) 679, 1053
PleI GAGTC 2 cut(s) 803, 1381
PpsI GAGTC 2 cut(s) 803, 1381
PspN4I GGNNCC 3 cut(s) 469, 557, 604
PspPI GGNCC 5 cut(s) 165, 468, 659, 830, 1320
PspXI VCTCGAGB 1 cut(s) 235
PstNI CAGNNNCTG 1 cut(s) 677
PvuII CAGCTG 1 cut(s) 446
RsaI GTAC 3 cut(s) 926, 1193, 1457
RsaNI GTAC 3 cut(s) 925, 1192, 1456
RseI CAYNNNNRTG 4 cut(s) 932, 1140, 1290, 1404
SaqAI TTAA 4 cut(s) 744, 1229, 1493, 1573
SatI GCNGC 2 cut(s) 678, 1052
Sau3AI GATC 5 cut(s) 18, 199, 449, 647, 1354
Sau96I GGNCC 5 cut(s) 165, 468, 659, 830, 1320
SchI GAGTC 2 cut(s) 804, 1381
SfaNI GCATC 8 cut(s) 307, 475, 768, 844, 926, 1015, 1144, 1408
SfcI CTRYAG 1 cut(s) 846
Sfr274I CTCGAG 2 cut(s) 235, 480
SinI GGWCC 2 cut(s) 468, 659
SlaI CTCGAG 2 cut(s) 235, 480
SmiMI CAYNNNNRTG 4 cut(s) 932, 1140, 1290, 1404
SmlI CTYRAG 7 cut(s) 235, 404, 480, 544, 956, 1228, 1492
SmoI CTYRAG 7 cut(s) 235, 404, 480, 544, 956, 1228, 1492
SpeI ACTAGT 1 cut(s) 782
Sse9I AATT 8 cut(s) 28, 120, 625, 1083, 1148, 1169, 1412, 1433
SsiI CCGC 1 cut(s) 851
SspI AATATT 1 cut(s) 1114
SspMI CTAG 5 cut(s) 540, 738, 783, 1205, 1469
StyI CCWWGG 3 cut(s) 84, 423, 607
TaaI ACNGT 6 cut(s) 193, 772, 826, 847, 951, 1348
TaiI ACGT 1 cut(s) 867
TaqI TCGA 6 cut(s) 180, 236, 481, 787, 972, 1335
TasI AATT 8 cut(s) 28, 120, 625, 1083, 1148, 1169, 1412, 1433
TatI WGTACW 1 cut(s) 924
TfiI GAWTC 7 cut(s) 268, 616, 1010, 1130, 1255, 1394, 1519
Tru1I TTAA 4 cut(s) 744, 1229, 1493, 1573
Tru9I TTAA 4 cut(s) 744, 1229, 1493, 1573
TscAI CASTG 7 cut(s) 53, 378, 775, 939, 1071, 1224, 1488
TseFI GTSAC 6 cut(s) 44, 694, 1199, 1283, 1463, 1547
TseI GCWGC 2 cut(s) 677, 1051
Tsp45I GTSAC 6 cut(s) 44, 694, 1199, 1283, 1463, 1547
TspDTI ATGAA 4 cut(s) 374, 562, 618, 629
TspGWI ACGGA 1 cut(s) 722
TspRI CASTG 7 cut(s) 53, 378, 775, 939, 1071, 1224, 1488
Vha464I CTTAAG 2 cut(s) 1228, 1492
VpaK11BI GGWCC 2 cut(s) 468, 659
XapI RAATTY 5 cut(s) 28, 1148, 1169, 1412, 1433
XhoI CTCGAG 2 cut(s) 235, 480
XmnI GAANNNNTTC 1 cut(s) 1122
XspI CTAG 5 cut(s) 540, 738, 783, 1205, 1469
Using CommOnly database (standard laboratory enzymes). Scanned on CDS sequence.