FvH4_6g01420

Neprosin

Design Primers CRISPR Design

Basic Information

Type: gene
Biological Identity
fragaria_vesca
Fvb6
OG0013071
Physical Location & Seq
Forward (+)
805929 .. 809969
4041 bp
Loading structure...
UTR
Exon/CDS
Intron
FvH4_6g01420.t2

Sequence Viewer

Length: 1335 bp
ATGAAAATGGATAGTCTCGTTCTACTTTTAGGCTTCCTTCTTCTCACTCTCTTCTCCCAAAGATTTTCGCTTCCGGTAGTGGTGGCCTTGGACCTGAAAAATACCACAAGGTACAATAAGCAAGTCAGCACCTTGAGACTTGAGAGAATTGAGAAGCACTTGAACAAGATCAACAAGCCTCCTGTGATGACTATACAGAGCCCAGATGGAGATATGATTGACTGTGTTCATAAAAGAAAGCAGCCAGCTCTGGATCACCCTCTCTTAAAAAACCACAAGATCCAGAAAGTGGCACCAGAAATGCCAAAAGTGATGAAGACGATGAAGCCATCTTCATCATCTTCTCAGAGTGAGAATGATACAGTGAAAGGCAAGACCGGTGGGATTGCATGGGGGCACCAAACGTGGCATACAAACGGGACGAGGTGTCCGAGAGGGACGGTTCCGATACGGCGGACTACAGTGAGTGATATTCTCAGAGCTAAGTCTTTGTTTGACTATGGCAAGAAAAGTCAACGACGCACCAATGTTAATGTGAATAGGCATGCTGATGCACCCGATGTGGTCAGTGGGAATGGCCATGAGCATGCGATTGCTTACTCCGGATCATCGAATGAAGTGTATGGTGCAAAAGCTACGATCAATGTTTGGACACCAGCAATCGAACAAATGAACGAGTTCAGTCTTTCGCAAATCTGGGTTCTGTCGGGATCATTCGATGGTACAGATCTCAACAGCATAGAAGCTGGATGGCAGGTCAGTCCGGAGCTCTATGGTGACAGCAGGCCTAGATTGTTCACCTATTGGACGACTGATTCATATCAGGCAACTGGATGCTACAACCTTCTGTGTTCTGGATTTGTGCAAACAAATAGTAGAATAGCCATCGGAGCTTCCATATCCCCGGTCTCATCGTATGCAAGCGACCAATTTGACGTTACTATCCTCATATGGAAGGATCCAAAGCTCGGAAATTGGTGGATGGGATATGGAGAGAATAATCTAGTAGGGTACTGGCCAGCAGCGCTATTCACACACTTAGCAGACAGAGCAACAATGGTGGAATGGGGTGGTGAGGTGGTCAATACAAGGTCCACCGGTGAGCATACCTCCACCCAAATGGGTTCTGGTCATTTTGCAGAAGACGGATTTGGGAAAGCAAGCTATTTCCGAAACCTAGAGATTGTAGACTCAGACAACAGCCTCAGTTCTGTGCGAGAAGTCTCAACCTTAGCTGAGAACACCAATTGTTACAACATCAAGAGCTCCTATAACAACGATTGGGGAACTTACTTCTACTATGGGGGGCCTGGAAGAAATCCAAAATGCCAATGA
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
Neprosin Neprosin_AP

Protein Analysis

445

Amino Acids

49.51

Weight (kDa)

8.7

Isoelectric Point (pI)

35.16

Instability Index
Protein Domains (Pfam)
Domain Name Pfam ID Position E-value Description
Neprosin_AP PF14365 64 - 200 2.1e-41 Neprosin activation peptide
Neprosin PF03080 225 - 437 2e-88 Neprosin
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: OG0013071)

Species Orthologous Gene IDs
arabidopsis_thaliana AT5G18460
fragaria_vesca FvH4_6g01420 FvH4_6g01420
malus_domestica MD04G1233600.v1.1
prunus_persica Prupe.6G353500_v2.0.a1 Prupe.6G353500_v2.0.a1
pyrus_communis pycom04g20750
rosa_chinensis RchiOBHm_Chr3g0449051
rosa_laevigata RLG00000025858
rosa_multiflora Rmu_co8511853.1_g000001 Rmu_sc0003116.1_g000008
rosa_roxburghii Rroxscaffold_6G00427160
rosa_rugosa Rorug02G0617700
rosa_samantha Rh3AG016100 Rh3BG016300 Rh3CG015300 Rh3DG016600
rosa_wichuraiana Rw3G001210

Restriction Enzyme Sites

1 / 10
Enzyme Recognition Site Cut Count Positions (bp)
Acc36I ACCTGC 1 cut(s) 745
AccB1I GGYRCC 2 cut(s) 292, 396
AccB7I CCANNNNNTGG 1 cut(s) 289
AccI GTMKAC 1 cut(s) 1188
AccIII TCCGGA 2 cut(s) 602, 763
AciI CCGC 1 cut(s) 454
AclWI GGATC 6 cut(s) 261, 274, 613, 718, 953, 966
AcoI YGGCCR 2 cut(s) 577, 1016
AfaI GTAC 3 cut(s) 113, 724, 1013
AfeI AGCGCT 1 cut(s) 1026
AfiI CCNNNNNNNGG 2 cut(s) 289, 968
AgeI ACCGGT 2 cut(s) 377, 1097
AgsI TTSAA 1 cut(s) 163
AhdI GACNNNNNGTC 1 cut(s) 426
AjnI CCWGG 1 cut(s) 1309
AjuI GAANNNNNNNTTGG 2 cut(s) 1134, 1166
Alw21I GWGCWC 2 cut(s) 771, 1268
Alw26I GTCTC 4 cut(s) 20, 130, 913, 1228
AlwI GGATC 6 cut(s) 261, 274, 613, 718, 953, 966
Aor13HI TCCGGA 2 cut(s) 602, 763
Aor51HI AGCGCT 1 cut(s) 1026
AoxI GGCC 5 cut(s) 84, 577, 785, 1016, 1307
ApeKI GCWGC 2 cut(s) 241, 1022
AsiGI ACCGGT 2 cut(s) 377, 1097
Asp700I GAANNNNTTC 1 cut(s) 677
AspLEI GCGC 1 cut(s) 1027
AspS9I GGNCC 3 cut(s) 91, 1092, 1307
AsuC2I CCSGG 1 cut(s) 905
AsuHPI GGTGA 5 cut(s) 248, 788, 790, 1085, 1112
AvaII GGWCC 2 cut(s) 91, 1092
BaeGI GKGCMC 1 cut(s) 399
BalI TGGCCA 2 cut(s) 579, 1018
BamHI GGATCC 1 cut(s) 958
BanI GGYRCC 2 cut(s) 292, 396
BanII GRGCYC 3 cut(s) 203, 771, 1268
BbsI GAAGAC 2 cut(s) 323, 1149
Bbv12I GWGCWC 2 cut(s) 771, 1268
BbvI GCAGC 2 cut(s) 253, 1034
BccI CCATC 6 cut(s) 200, 337, 713, 744, 893, 976
BceAI ACGGC 1 cut(s) 467
BciT130I CCWGG 1 cut(s) 1311
BcnI CCSGG 1 cut(s) 905
BcoDI GTCTC 4 cut(s) 20, 130, 913, 1228
BfaI CTAG 3 cut(s) 789, 1004, 1178
BfmI CTRYAG 1 cut(s) 459
BfoI RGCGCY 1 cut(s) 1028
BfuAI ACCTGC 1 cut(s) 745
BglII AGATCT 1 cut(s) 727
BisI GCNGC 2 cut(s) 242, 1023
BlsI GCNGC 2 cut(s) 243, 1024
Bme1390I CCNGG 2 cut(s) 905, 1311
Bme18I GGWCC 2 cut(s) 91, 1092
BmeRI GACNNNNNGTC 1 cut(s) 426
BmgT120I GGNCC 3 cut(s) 91, 1092, 1307
BmiI GGNNCC 5 cut(s) 294, 398, 444, 960, 1308
BmrFI CCNGG 2 cut(s) 905, 1311
BmsI GCATC 2 cut(s) 541, 824
BpiI GAAGAC 2 cut(s) 323, 1149
BplI GAGNNNNNCTC 2 cut(s) 1094, 1126
Bpu10I CCTNAGC 1 cut(s) 1231
BpuEI CTTGAG 2 cut(s) 154, 161
BpuMI CCSGG 1 cut(s) 905
BsaBI GATNNNNATC 1 cut(s) 819
BsaI GGTCTC 1 cut(s) 913
BsaJI CCNNGG 2 cut(s) 87, 903
BsaWI WCCGGW 5 cut(s) 73, 377, 602, 763, 1097
Bsc4I CCNNNNNNNGG 2 cut(s) 289, 968
Bse118I RCCGGY 2 cut(s) 377, 1097
Bse1I ACTGG 2 cut(s) 835, 1019
Bse8I GATNNNNATC 1 cut(s) 819
BseAI TCCGGA 2 cut(s) 602, 763
BseBI CCWGG 1 cut(s) 1311
BseDI CCNNGG 2 cut(s) 87, 903
BseGI GGATG 3 cut(s) 755, 839, 987
BseJI GATNNNNATC 1 cut(s) 819
BseLI CCNNNNNNNGG 2 cut(s) 289, 968
BseMII CTCAG 5 cut(s) 359, 490, 1206, 1219, 1227
BseNI ACTGG 2 cut(s) 835, 1019
BseSI GKGCMC 1 cut(s) 399
BseXI GCAGC 2 cut(s) 253, 1034
BshFI GGCC 5 cut(s) 86, 579, 787, 1018, 1309
BshNI GGYRCC 2 cut(s) 292, 396
BshTI ACCGGT 2 cut(s) 377, 1097
BsiHKAI GWGCWC 2 cut(s) 771, 1268
BsiSI CCGG 6 cut(s) 74, 378, 603, 764, 905, 1098
BslFI GGGAC 2 cut(s) 433, 451
BslI CCNNNNNNNGG 2 cut(s) 289, 968
BsmAI GTCTC 4 cut(s) 20, 130, 913, 1228
BsmFI GGGAC 2 cut(s) 433, 451
BsnI GGCC 5 cut(s) 86, 579, 787, 1018, 1309
Bso31I GGTCTC 1 cut(s) 913
Bsp1286I GDGCHC 4 cut(s) 203, 399, 771, 1268
Bsp13I TCCGGA 2 cut(s) 602, 763
Bsp143I GATC 8 cut(s) 168, 253, 279, 605, 639, 710, 727, 958
BspACI CCGC 1 cut(s) 454
BspANI GGCC 5 cut(s) 86, 579, 787, 1018, 1309
BspCNI CTCAG 5 cut(s) 358, 489, 1205, 1218, 1228
BspEI TCCGGA 2 cut(s) 602, 763
BspLI GGNNCC 5 cut(s) 294, 398, 444, 960, 1308
BspMI ACCTGC 1 cut(s) 745
BspPI GGATC 6 cut(s) 261, 274, 613, 718, 953, 966
BspT107I GGYRCC 2 cut(s) 292, 396
BspTNI GGTCTC 1 cut(s) 913
BsrFI RCCGGY 2 cut(s) 377, 1097
BsrI ACTGG 2 cut(s) 835, 1019
BssAI RCCGGY 2 cut(s) 377, 1097
BssECI CCNNGG 2 cut(s) 87, 903
BssMI GATC 8 cut(s) 168, 253, 279, 605, 639, 710, 727, 958
BssT1I CCWWGG 1 cut(s) 87
Bst2UI CCWGG 1 cut(s) 1311
Bst4CI ACNGT 4 cut(s) 224, 364, 442, 463
Bst6I CTCTTC 1 cut(s) 56
BstC8I GCNNGC 7 cut(s) 246, 546, 588, 785, 922, 1020, 1162
BstDEI CTNAG 8 cut(s) 345, 476, 483, 1039, 1192, 1205, 1231, 1236
BstF5I GGATG 3 cut(s) 755, 839, 987
BstH2I RGCGCY 1 cut(s) 1028
BstHHI GCGC 1 cut(s) 1027
BstKTI GATC 8 cut(s) 171, 256, 282, 608, 642, 713, 730, 961
BstMAI GTCTC 4 cut(s) 20, 130, 913, 1228
BstMBI GATC 8 cut(s) 168, 253, 279, 605, 639, 710, 727, 958
BstMWI GCNNNNNNNGC 3 cut(s) 890, 1024, 1049
BstNI CCWGG 1 cut(s) 1311
BstNSI RCATGY 2 cut(s) 548, 590
BstSCI CCNGG 2 cut(s) 903, 1309
BstSFI CTRYAG 1 cut(s) 459
BstSLI GKGCMC 1 cut(s) 399
BstV1I GCAGC 2 cut(s) 253, 1034
BstV2I GAAGAC 2 cut(s) 323, 1149
BstX2I RGATCY 3 cut(s) 279, 727, 958
BstXI CCANNNNNNTGG 1 cut(s) 1120
BstYI RGATCY 3 cut(s) 279, 727, 958
BsuRI GGCC 5 cut(s) 86, 579, 787, 1018, 1309
BtsCI GGATG 3 cut(s) 755, 839, 987
BtsIMutI CAGTG 3 cut(s) 369, 468, 574
BveI ACCTGC 1 cut(s) 745
Cac8I GCNNGC 7 cut(s) 246, 546, 588, 785, 922, 1020, 1162
CfoI GCGC 1 cut(s) 1027
Cfr10I RCCGGY 2 cut(s) 377, 1097
Cfr13I GGNCC 3 cut(s) 91, 1092, 1307
CseI GACGC 1 cut(s) 528
Csp6I GTAC 3 cut(s) 112, 723, 1012
CspAI ACCGGT 2 cut(s) 377, 1097
CspCI CAANNNNNGTGG 4 cut(s) 361, 396, 1041, 1076
CviAII CATG 4 cut(s) 390, 545, 581, 587
CviQI GTAC 3 cut(s) 112, 723, 1012
DdeI CTNAG 8 cut(s) 345, 476, 483, 1039, 1192, 1205, 1231, 1236
DpnI GATC 8 cut(s) 170, 255, 281, 607, 641, 712, 729, 960
DpnII GATC 8 cut(s) 168, 253, 279, 605, 639, 710, 727, 958
DriI GACNNNNNGTC 1 cut(s) 426
EaeI YGGCCR 2 cut(s) 577, 1016
Eam1104I CTCTTC 1 cut(s) 56
Eam1105I GACNNNNNGTC 1 cut(s) 426
EarI CTCTTC 1 cut(s) 56
EciI GGCGGA 1 cut(s) 469
Ecl136II GAGCTC 2 cut(s) 769, 1266
Eco130I CCWWGG 1 cut(s) 87
Eco147I AGGCCT 1 cut(s) 787
Eco24I GRGCYC 3 cut(s) 203, 771, 1268
Eco31I GGTCTC 1 cut(s) 913
Eco47I GGWCC 2 cut(s) 91, 1092
Eco47III AGCGCT 1 cut(s) 1026
Eco53kI GAGCTC 2 cut(s) 769, 1266
EcoICRI GAGCTC 2 cut(s) 769, 1266
EcoO109I RGGNCCY 1 cut(s) 1307
EcoRII CCWGG 1 cut(s) 1309
EcoT14I CCWWGG 1 cut(s) 87
EcoT38I GRGCYC 3 cut(s) 203, 771, 1268
ErhI CCWWGG 1 cut(s) 87
FaeI CATG 4 cut(s) 393, 548, 584, 590
FaqI GGGAC 2 cut(s) 433, 451
FatI CATG 4 cut(s) 389, 544, 580, 586
FauNDI CATATG 1 cut(s) 950
FblI GTMKAC 1 cut(s) 1188
Fnu4HI GCNGC 2 cut(s) 242, 1023
FokI GGATG 3 cut(s) 762, 846, 994
FriOI GRGCYC 3 cut(s) 203, 771, 1268
Fsp4HI GCNGC 2 cut(s) 242, 1023
FspBI CTAG 3 cut(s) 789, 1004, 1178
GlaI GCGC 1 cut(s) 1026
GluI GCNGC 2 cut(s) 242, 1023
HaeII RGCGCY 1 cut(s) 1028
HaeIII GGCC 5 cut(s) 86, 579, 787, 1018, 1309
HapII CCGG 6 cut(s) 74, 378, 603, 764, 905, 1098
HgaI GACGC 1 cut(s) 528
HhaI GCGC 1 cut(s) 1027
Hin1II CATG 4 cut(s) 393, 548, 584, 590
Hin6I GCGC 1 cut(s) 1025
HinP1I GCGC 1 cut(s) 1025
HincII GTYRAC 1 cut(s) 515
HindII GTYRAC 1 cut(s) 515
HinfI GANTC 2 cut(s) 815, 1190
HpaII CCGG 6 cut(s) 74, 378, 603, 764, 905, 1098
HphI GGTGA 5 cut(s) 248, 788, 790, 1085, 1112
Hpy166II GTNNAC 4 cut(s) 515, 798, 1095, 1189
Hpy188I TCNGA 8 cut(s) 348, 432, 447, 479, 890, 971, 1172, 1195
Hpy188III TCNNGA 7 cut(s) 251, 283, 603, 708, 764, 855, 1261
Hpy8I GTNNAC 4 cut(s) 515, 798, 1095, 1189
Hpy99I CGWCG 1 cut(s) 522
HpyAV CCTTC 3 cut(s) 47, 854, 949
HpyCH4III ACNGT 4 cut(s) 224, 364, 442, 463
HpyCH4IV ACGT 2 cut(s) 404, 936
HpyCH4V TGCA 6 cut(s) 389, 554, 629, 865, 920, 1139
HpyF10VI GCNNNNNNNGC 3 cut(s) 890, 1024, 1049
HpyF3I CTNAG 8 cut(s) 345, 476, 483, 1039, 1192, 1205, 1231, 1236
HpySE526I ACGT 2 cut(s) 404, 936
Hsp92II CATG 4 cut(s) 393, 548, 584, 590
HspAI GCGC 1 cut(s) 1025
Kpn2I TCCGGA 2 cut(s) 602, 763
Kzo9I GATC 8 cut(s) 168, 253, 279, 605, 639, 710, 727, 958
LmnI GCTCC 3 cut(s) 766, 890, 1271
Lsp1109I GCAGC 2 cut(s) 253, 1034
LweI GCATC 2 cut(s) 541, 824
MaeI CTAG 3 cut(s) 789, 1004, 1178
MaeII ACGT 2 cut(s) 404, 936
MaeIII GTNAC 3 cut(s) 776, 937, 1250
MalI GATC 8 cut(s) 170, 255, 281, 607, 641, 712, 729, 960
MboI GATC 8 cut(s) 168, 253, 279, 605, 639, 710, 727, 958
MboII GAAGA 7 cut(s) 32, 43, 324, 328, 333, 1154, 1326
MfeI CAATTG 1 cut(s) 1246
MflI RGATCY 3 cut(s) 279, 727, 958
MhlI GDGCHC 4 cut(s) 203, 399, 771, 1268
MlsI TGGCCA 2 cut(s) 579, 1018
MluCI AATT 4 cut(s) 147, 929, 973, 1246
MluNI TGGCCA 2 cut(s) 579, 1018
MlyI GAGTC 1 cut(s) 1184
MnlI CCTC 8 cut(s) 189, 270, 417, 428, 956, 1069, 1120, 1214
Mox20I TGGCCA 2 cut(s) 579, 1018
MroI TCCGGA 2 cut(s) 602, 763
MroXI GAANNNNTTC 1 cut(s) 677
MscI TGGCCA 2 cut(s) 579, 1018
MseI TTAA 2 cut(s) 266, 531
MslI CAYNNNNRTG 3 cut(s) 549, 585, 1118
Msp20I TGGCCA 2 cut(s) 579, 1018
MspI CCGG 6 cut(s) 74, 378, 603, 764, 905, 1098
MspR9I CCNGG 2 cut(s) 905, 1311
MunI CAATTG 1 cut(s) 1246
MvaI CCWGG 1 cut(s) 1311
MwoI GCNNNNNNNGC 3 cut(s) 890, 1024, 1049
NciI CCSGG 1 cut(s) 905
NdeI CATATG 1 cut(s) 950
NdeII GATC 8 cut(s) 168, 253, 279, 605, 639, 710, 727, 958
NlaIII CATG 4 cut(s) 393, 548, 584, 590
NlaIV GGNNCC 5 cut(s) 294, 398, 444, 960, 1308
NmuCI GTSAC 1 cut(s) 776
NspI RCATGY 2 cut(s) 548, 590
PaeI GCATGC 2 cut(s) 548, 590
PceI AGGCCT 1 cut(s) 787
PcsI WCGNNNNNNNCGW 1 cut(s) 428
PdmI GAANNNNTTC 1 cut(s) 677
PfeI GAWTC 1 cut(s) 815
PflMI CCANNNNNTGG 1 cut(s) 289
PinAI ACCGGT 2 cut(s) 377, 1097
PkrI GCNGC 2 cut(s) 243, 1024
PleI GAGTC 1 cut(s) 1184
PpsI GAGTC 1 cut(s) 1184
Psp124BI GAGCTC 2 cut(s) 771, 1268
Psp6I CCWGG 1 cut(s) 1309
PspGI CCWGG 1 cut(s) 1309
PspN4I GGNNCC 5 cut(s) 294, 398, 444, 960, 1308
PspPI GGNCC 3 cut(s) 91, 1092, 1307
PsuI RGATCY 3 cut(s) 279, 727, 958
RsaI GTAC 3 cut(s) 113, 724, 1013
RsaNI GTAC 3 cut(s) 112, 723, 1012
RseI CAYNNNNRTG 3 cut(s) 549, 585, 1118
SacI GAGCTC 2 cut(s) 771, 1268
SaqAI TTAA 2 cut(s) 266, 531
SatI GCNGC 2 cut(s) 242, 1023
Sau3AI GATC 8 cut(s) 168, 253, 279, 605, 639, 710, 727, 958
Sau96I GGNCC 3 cut(s) 91, 1092, 1307
SchI GAGTC 1 cut(s) 1184
ScrFI CCNGG 2 cut(s) 905, 1311
SduI GDGCHC 4 cut(s) 203, 399, 771, 1268
SfaNI GCATC 2 cut(s) 541, 824
SfcI CTRYAG 1 cut(s) 459
SgrAI CRCCGGYG 1 cut(s) 1097
SinI GGWCC 2 cut(s) 91, 1092
SmiMI CAYNNNNRTG 3 cut(s) 549, 585, 1118
SmlI CTYRAG 2 cut(s) 133, 140
SmoI CTYRAG 2 cut(s) 133, 140
SphI GCATGC 2 cut(s) 548, 590
Sse9I AATT 4 cut(s) 147, 929, 973, 1246
SseBI AGGCCT 1 cut(s) 787
SsiI CCGC 1 cut(s) 454
SspMI CTAG 3 cut(s) 789, 1004, 1178
SstI GAGCTC 2 cut(s) 771, 1268
StuI AGGCCT 1 cut(s) 787
StyD4I CCNGG 2 cut(s) 903, 1309
StyI CCWWGG 1 cut(s) 87
TaaI ACNGT 4 cut(s) 224, 364, 442, 463
TaiI ACGT 2 cut(s) 407, 939
TaqI TCGA 3 cut(s) 611, 663, 717
TasI AATT 4 cut(s) 147, 929, 973, 1246
TfiI GAWTC 1 cut(s) 815
Tru1I TTAA 2 cut(s) 266, 531
Tru9I TTAA 2 cut(s) 266, 531
TscAI CASTG 3 cut(s) 369, 468, 574
TseFI GTSAC 1 cut(s) 776
TseI GCWGC 2 cut(s) 241, 1022
Tsp45I GTSAC 1 cut(s) 776
TspDTI ATGAA 8 cut(s) 17, 218, 324, 329, 338, 630, 686, 807
TspGWI ACGGA 1 cut(s) 1161
TspRI CASTG 3 cut(s) 369, 468, 574
Van91I CCANNNNNTGG 1 cut(s) 289
VpaK11BI GGWCC 2 cut(s) 91, 1092
XceI RCATGY 2 cut(s) 548, 590
XcmI CCANNNNNNNNNTGG 1 cut(s) 1124
XmiI GTMKAC 1 cut(s) 1188
XmnI GAANNNNTTC 1 cut(s) 677
XspI CTAG 3 cut(s) 789, 1004, 1178
Using CommOnly database (standard laboratory enzymes). Scanned on CDS sequence.