Rh6DG484000

3-oxo-Delta(4,5)-steroid 5-beta-reductase-like

Design Primers CRISPR Design

Basic Information

Type: gene
Biological Identity
rosa_samantha
Chr6D
OG0000626
Physical Location & Seq
Reverse (-)
64674848 .. 64676527
1680 bp
Loading structure...
UTR
Exon/CDS
Intron
Rh6DG484000.1

Sequence Viewer

Length: 1173 bp
ATGAGCTGGTGGTGGGCCGGAGCCGTTGGCACTGCCAAGAGAAAATTAGAAGAAGATGAAGCGCCGCGAACGCCGCAGAGCGTGGGGTTAGTGATCGGCGTGACGGGAATAGTCGGCAACAGCCTCGCCGAGATCCTCCCTCTCTCCGACACCCCCGGCGGTCCATGGAAGGTCTACGGCGTCGCCCGCCGTCCCCGGCCCAACTGGAACGCCGATCACCCCGTCGAGTACATCCAGTGCGACGTCTCCGACGCCGAGGACGCCAAGGCCAAGCTCACGCCGTTGACTGACGTCACTCACCTCTTCTACGTCACCTGGTCCAGCCGATCCACCGAGGCCGAGAATTGCGAGGCCAACGCCGTGATGCTGCGCAACGTTTTGAGCTCCGTGATCCCAATCGCACCGAATCTGAAGCACATATGCCTCCAGACCGGAGGCAAACATTACATCGGGCCGTTCGAGTCGTGGGGGAAGATCGAGCCGCACGACACGCCGTTCACAGAGGATCTGCCCCGGTTGAGCTACCAGAATTTCTACTACACACTCGAAGATATATTGTTCGAAGAGGTGGAGAAGAAAGATGAGCTAACTTGGTCGGTGCACCGACCCGACACCATTTTCGGGTTTTCGCCTTATAGCTTGATGAACATTGTAGGCACTCTCTGTGTGTATGCTGCTATATGCAAACATGAGGGCTTGCCACTGAGATTTCCGGGAACCAAAGAGGCGTGGAATTGCTTGTCGGCAGCTTCGGATGCGGATTTGATAGCGGAGCAGCAGATATGGGCTGCAGTGGATCCTTATGCTCGAAACGAAGCCTTCAATATCAACAATGCGGATGTGTTCAGGTGGAAGCATTTCTGGAAGGTGTTGGCTGAGAATTTCGGGATAGAGGAATTCGGGTTTGAGGAGAAGGGGGAGAGGTTGAGCTTGGAGAAGTTGATGGAGGGGAAGAATGGAGTGTGGGAGGAGATAGTGAAGGAGAATCAGCTGCTGCCGACGAAGCTGGAGGAGGTTGGAGTGTGGTGGTTTGTGGATACTGTGTTTAGTGGGGAAAGTAAGTTTGGTTGTATGAACAAGAGTAAAGAACATGGGTTTGTGGGGTTTAGGAACTCCAGGAATTCCTTTGTTACTTGGATTGATAAAATGAAAGCTTACAAGCTTGTGCCTTGA

Protein Analysis

390

Amino Acids

44.25

Weight (kDa)

5.22

Isoelectric Point (pI)

39.99

Instability Index
Protein Domains (Pfam)
Domain Name Pfam ID Position E-value Description
PRISE PF22917 87 - 390 5.5e-29 PRISE-like Rossmann-fold 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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Orthologous Genes (Group: OG0000626)

Species Orthologous Gene IDs
arabidopsis_thaliana AT4G24220 AT4G24220
fragaria_vesca FvH4_2g04580 FvH4_2g04580 FvH4_2g04580 FvH4_4g06071 FvH4_5g25130
malus_domestica MD05G1094500.v1.1 MD05G1108300.v1.1 MD08G1088400.v1.1 MD15G1073200.v1.1 MD15G1073300.v1.1 MD15G1073400.v1.1 MD15G1073500.v1.1 MD15G1073600.v1.1
prunus_persica Prupe.1G426100_v2.0.a1 Prupe.1G426100_v2.0.a1 Prupe.1G426200_v2.0.a1 Prupe.1G426300_v2.0.a1 Prupe.1G426400_v2.0.a1 Prupe.1G426400_v2.0.a1 Prupe.8G139400_v2.0.a1 Prupe.8G139400_v2.0.a1
pyrus_communis pycom05g09350 pycom08g07190 pycom13g22780 pycom15g06840 pycom15g06850 pycom15g06870
rosa_chinensis RchiOBHm_Chr6g0254161 RchiOBHm_Chr6g0254181 RchiOBHm_Chr6g0254191 RchiOBHm_Chr6g0309171 RchiOBHm_Chr6g0309181 RchiOBHm_Chr6g0309191 RchiOBHm_Chr6g0309201
rosa_laevigata RLG00000010582 RLG00000010583 RLG00000014937 RLG00000014938
rosa_multiflora Rmu_co8060540.1_g000001 Rmu_co8332663.1_g000001 Rmu_sc0000010.1_g000017 Rmu_sc0003711.1_g000008 Rmu_sc0016971.1_g000003 Rmu_sc0020102.1_g000006 Rmu_sc0020102.1_g000008
rosa_roxburghii Rroxscaffold_7G00159570 Rroxscaffold_7G00159580 Rroxscaffold_7G00159590 Rroxscaffold_7G00211100
rosa_rugosa Rorug05G0550400 Rorug06G0370100 Rorug06G0370300
rosa_samantha Rh6AG066900 Rh6AG483200 Rh6AG483300 Rh6AG483400 Rh6BG059800 Rh6BG060000 Rh6BG492200 Rh6BG492300 Rh6BG492500 Rh6CG059700 Rh6CG059800 Rh6CG496900 Rh6CG497000 Rh6CG497200 Rh6CG497400 Rh6DG056700 Rh6DG056800 Rh6DG483800 Rh6DG484000
rosa_wichuraiana Rw6G005860 Rw6G005870 Rw6G042070 Rw6G042080 Rw6G042090

Restriction Enzyme Sites

1 / 10
Enzyme Recognition Site Cut Count Positions (bp)
AatII GACGTC 2 cut(s) 246, 294
Acc16I TGCGCA 1 cut(s) 371
AccI GTMKAC 1 cut(s) 174
AccII CGCG 1 cut(s) 67
AciI CCGC 8 cut(s) 65, 74, 159, 187, 482, 758, 770, 836
AclI AACGTT 1 cut(s) 375
AclWI GGATC 6 cut(s) 127, 321, 385, 513, 791, 804
AcsI RAATTY 4 cut(s) 529, 880, 896, 1120
AcuI CTGAAG 1 cut(s) 431
AcyI GRCGYC 5 cut(s) 180, 243, 252, 261, 291
AfaI GTAC 1 cut(s) 230
AfiI CCNNNNNNNGG 1 cut(s) 621
AgsI TTSAA 1 cut(s) 823
AjnI CCWGG 2 cut(s) 314, 1115
AjuI GAANNNNNNNTTGG 2 cut(s) 1047, 1079
Alw21I GWGCWC 2 cut(s) 386, 603
Alw26I GTCTC 1 cut(s) 250
Alw44I GTGCAC 1 cut(s) 599
AlwI GGATC 6 cut(s) 127, 321, 385, 513, 791, 804
AlwNI CAGNNNCTG 1 cut(s) 994
AoxI GGCC 6 cut(s) 15, 197, 267, 336, 351, 452
ApaLI GTGCAC 1 cut(s) 599
ApeKI GCWGC 7 cut(s) 367, 674, 746, 775, 788, 991, 994
ApoI RAATTY 4 cut(s) 529, 880, 896, 1120
ArsI GACNNNNNNTTYG 2 cut(s) 602, 634
Asp700I GAANNNNTTC 1 cut(s) 857
AspLEI GCGC 2 cut(s) 64, 372
AspS9I GGNCC 5 cut(s) 15, 161, 198, 318, 452
AsuC2I CCSGG 4 cut(s) 156, 196, 514, 714
AsuHPI GGTGA 3 cut(s) 209, 290, 304
AsuII TTCGAA 1 cut(s) 561
AvaII GGWCC 2 cut(s) 161, 318
BaeGI GKGCMC 1 cut(s) 603
BamHI GGATCC 1 cut(s) 796
BanII GRGCYC 1 cut(s) 386
Bbv12I GWGCWC 2 cut(s) 386, 603
BbvI GCAGC 7 cut(s) 354, 661, 758, 775, 787, 978, 981
BccI CCATC 1 cut(s) 937
BceAI ACGGC 7 cut(s) 8, 174, 193, 265, 344, 439, 478
BcgI CGANNNNNNTGC 2 cut(s) 106, 140
BciT130I CCWGG 2 cut(s) 316, 1117
BciVI GTATCC 1 cut(s) 1030
BcnI CCSGG 4 cut(s) 156, 196, 514, 714
BcoDI GTCTC 1 cut(s) 250
BfmI CTRYAG 1 cut(s) 789
BfoI RGCGCY 1 cut(s) 65
BfuI GTATCC 1 cut(s) 1030
Bme1390I CCNGG 6 cut(s) 156, 196, 316, 514, 714, 1117
Bme18I GGWCC 2 cut(s) 161, 318
BmgT120I GGNCC 5 cut(s) 15, 161, 198, 318, 452
BmiI GGNNCC 3 cut(s) 22, 718, 798
BmrFI CCNGG 6 cut(s) 156, 196, 316, 514, 714, 1117
BmsI GCATC 2 cut(s) 354, 745
BoxI GACNNNNGTC 1 cut(s) 290
BpmI CTGGAG 3 cut(s) 410, 1028, 1099
Bpu14I TTCGAA 1 cut(s) 561
BpuMI CCSGG 4 cut(s) 156, 196, 514, 714
BsaBI GATNNNNATC 1 cut(s) 395
BsaHI GRCGYC 5 cut(s) 180, 243, 252, 261, 291
BsaJI CCNNGG 7 cut(s) 154, 164, 194, 255, 264, 333, 512
BsaWI WCCGGW 1 cut(s) 431
Bsc4I CCNNNNNNNGG 1 cut(s) 621
Bse1I ACTGG 2 cut(s) 209, 235
Bse8I GATNNNNATC 1 cut(s) 395
BseBI CCWGG 2 cut(s) 316, 1117
BseDI CCNNGG 7 cut(s) 154, 164, 194, 255, 264, 333, 512
BseGI GGATG 3 cut(s) 231, 760, 844
BseJI GATNNNNATC 1 cut(s) 395
BseLI CCNNNNNNNGG 1 cut(s) 621
BseMII CTCAG 2 cut(s) 695, 867
BseNI ACTGG 2 cut(s) 209, 235
BseRI GAGGAG 3 cut(s) 923, 983, 1025
BseSI GKGCMC 1 cut(s) 603
BseXI GCAGC 7 cut(s) 354, 661, 758, 775, 787, 978, 981
Bsh1236I CGCG 1 cut(s) 67
BshFI GGCC 6 cut(s) 17, 199, 269, 338, 353, 454
BsiHKAI GWGCWC 2 cut(s) 386, 603
BsiSI CCGG 6 cut(s) 18, 156, 196, 432, 514, 713
BslFI GGGAC 1 cut(s) 177
BslI CCNNNNNNNGG 1 cut(s) 621
BsmAI GTCTC 1 cut(s) 250
BsmBI CGTCTC 1 cut(s) 250
BsmFI GGGAC 1 cut(s) 177
BsnI GGCC 6 cut(s) 17, 199, 269, 338, 353, 454
Bsp119I TTCGAA 1 cut(s) 561
Bsp1286I GDGCHC 2 cut(s) 386, 603
Bsp143I GATC 8 cut(s) 93, 132, 214, 326, 390, 474, 505, 796
Bsp19I CCATGG 1 cut(s) 164
BspACI CCGC 8 cut(s) 65, 74, 159, 187, 482, 758, 770, 836
BspANI GGCC 6 cut(s) 17, 199, 269, 338, 353, 454
BspCNI CTCAG 2 cut(s) 696, 868
BspFNI CGCG 1 cut(s) 67
BspLI GGNNCC 3 cut(s) 22, 718, 798
BspMAI CTGCAG 1 cut(s) 793
BspPI GGATC 6 cut(s) 127, 321, 385, 513, 791, 804
BspT104I TTCGAA 1 cut(s) 561
BsrI ACTGG 2 cut(s) 209, 235
BssECI CCNNGG 7 cut(s) 154, 164, 194, 255, 264, 333, 512
BssMI GATC 8 cut(s) 93, 132, 214, 326, 390, 474, 505, 796
BssNI GRCGYC 5 cut(s) 180, 243, 252, 261, 291
BssT1I CCWWGG 2 cut(s) 164, 264
Bst2UI CCWGG 2 cut(s) 316, 1117
Bst4CI ACNGT 1 cut(s) 1042
Bst6I CTCTTC 2 cut(s) 308, 558
BstACI GRCGYC 5 cut(s) 180, 243, 252, 261, 291
BstBI TTCGAA 1 cut(s) 561
BstC8I GCNNGC 2 cut(s) 187, 698
BstDEI CTNAG 2 cut(s) 704, 876
BstDSI CCRYGG 1 cut(s) 164
BstF5I GGATG 3 cut(s) 231, 760, 844
BstFNI CGCG 1 cut(s) 67
BstH2I RGCGCY 1 cut(s) 65
BstHHI GCGC 2 cut(s) 64, 372
BstKTI GATC 8 cut(s) 96, 135, 217, 329, 393, 477, 508, 799
BstMAI GTCTC 1 cut(s) 250
BstMBI GATC 8 cut(s) 93, 132, 214, 326, 390, 474, 505, 796
BstMWI GCNNNNNNNGC 7 cut(s) 70, 73, 186, 260, 490, 755, 1003
BstNI CCWGG 2 cut(s) 316, 1117
BstPAI GACNNNNGTC 1 cut(s) 290
BstSCI CCNGG 6 cut(s) 154, 194, 314, 512, 712, 1115
BstSFI CTRYAG 1 cut(s) 789
BstSLI GKGCMC 1 cut(s) 603
BstUI CGCG 1 cut(s) 67
BstV1I GCAGC 7 cut(s) 354, 661, 758, 775, 787, 978, 981
BstX2I RGATCY 3 cut(s) 132, 505, 796
BstYI RGATCY 3 cut(s) 132, 505, 796
BsuI GTATCC 1 cut(s) 1030
BsuRI GGCC 6 cut(s) 17, 199, 269, 338, 353, 454
BtgI CCRYGG 1 cut(s) 164
BtsCI GGATG 3 cut(s) 231, 760, 844
BtsI GCAGTG 2 cut(s) 30, 798
BtsIMutI CAGTG 4 cut(s) 30, 242, 701, 798
Cac8I GCNNGC 2 cut(s) 187, 698
CaiI CAGNNNCTG 1 cut(s) 994
CfoI GCGC 2 cut(s) 64, 372
Cfr13I GGNCC 5 cut(s) 15, 161, 198, 318, 452
CseI GACGC 3 cut(s) 169, 260, 269
CsiI ACCWGGT 1 cut(s) 314
Csp6I GTAC 1 cut(s) 229
CviAII CATG 3 cut(s) 165, 689, 1091
CviQI GTAC 1 cut(s) 229
DdeI CTNAG 2 cut(s) 704, 876
DpnI GATC 8 cut(s) 95, 134, 216, 328, 392, 476, 507, 798
DpnII GATC 8 cut(s) 93, 132, 214, 326, 390, 474, 505, 796
Eam1104I CTCTTC 2 cut(s) 308, 558
EarI CTCTTC 2 cut(s) 308, 558
Ecl136II GAGCTC 1 cut(s) 384
Eco130I CCWWGG 2 cut(s) 164, 264
Eco24I GRGCYC 1 cut(s) 386
Eco47I GGWCC 2 cut(s) 161, 318
Eco53kI GAGCTC 1 cut(s) 384
Eco57I CTGAAG 1 cut(s) 431
EcoICRI GAGCTC 1 cut(s) 384
EcoRI GAATTC 2 cut(s) 896, 1120
EcoRII CCWGG 2 cut(s) 314, 1115
EcoT14I CCWWGG 2 cut(s) 164, 264
EcoT38I GRGCYC 1 cut(s) 386
ErhI CCWWGG 2 cut(s) 164, 264
Esp3I CGTCTC 1 cut(s) 250
FaeI CATG 3 cut(s) 168, 692, 1094
FaqI GGGAC 1 cut(s) 177
FatI CATG 3 cut(s) 164, 688, 1090
FauI CCCGC 1 cut(s) 194
FauNDI CATATG 1 cut(s) 419
FblI GTMKAC 1 cut(s) 174
FokI GGATG 3 cut(s) 218, 767, 851
FriOI GRGCYC 1 cut(s) 386
FspI TGCGCA 1 cut(s) 371
GlaI GCGC 2 cut(s) 63, 371
GsuI CTGGAG 3 cut(s) 410, 1028, 1099
HaeII RGCGCY 1 cut(s) 65
HaeIII GGCC 6 cut(s) 17, 199, 269, 338, 353, 454
HapII CCGG 6 cut(s) 18, 156, 196, 432, 514, 713
HgaI GACGC 3 cut(s) 169, 260, 269
HhaI GCGC 2 cut(s) 64, 372
Hin1I GRCGYC 5 cut(s) 180, 243, 252, 261, 291
Hin1II CATG 3 cut(s) 168, 692, 1094
Hin6I GCGC 2 cut(s) 62, 370
HinP1I GCGC 2 cut(s) 62, 370
HincII GTYRAC 1 cut(s) 285
HindII GTYRAC 1 cut(s) 285
HindIII AAGCTT 2 cut(s) 1152, 1160
HinfI GANTC 3 cut(s) 406, 461, 985
HpaII CCGG 6 cut(s) 18, 156, 196, 432, 514, 713
HphI GGTGA 3 cut(s) 209, 290, 304
Hpy166II GTNNAC 4 cut(s) 175, 285, 498, 601
Hpy188I TCNGA 4 cut(s) 148, 250, 411, 754
Hpy188III TCNNGA 3 cut(s) 427, 862, 886
Hpy8I GTNNAC 4 cut(s) 175, 285, 498, 601
Hpy99I CGWCG 5 cut(s) 185, 227, 245, 254, 1003
HpyAV CCTTC 5 cut(s) 163, 829, 859, 907, 973
HpyCH4III ACNGT 1 cut(s) 1042
HpyCH4IV ACGT 4 cut(s) 243, 291, 309, 375
HpyCH4V TGCA 3 cut(s) 601, 684, 791
HpyF10VI GCNNNNNNNGC 7 cut(s) 70, 73, 186, 260, 490, 755, 1003
HpyF3I CTNAG 2 cut(s) 704, 876
HpySE526I ACGT 4 cut(s) 243, 291, 309, 375
Hsp92I GRCGYC 5 cut(s) 180, 243, 252, 261, 291
Hsp92II CATG 3 cut(s) 168, 692, 1094
HspAI GCGC 2 cut(s) 62, 370
Kzo9I GATC 8 cut(s) 93, 132, 214, 326, 390, 474, 505, 796
LmnI GCTCC 3 cut(s) 20, 389, 772
Lsp1109I GCAGC 7 cut(s) 354, 661, 758, 775, 787, 978, 981
LweI GCATC 2 cut(s) 354, 745
MabI ACCWGGT 1 cut(s) 314
MaeII ACGT 4 cut(s) 243, 291, 309, 375
MaeIII GTNAC 4 cut(s) 100, 292, 310, 1129
MalI GATC 8 cut(s) 95, 134, 216, 328, 392, 476, 507, 798
MboI GATC 8 cut(s) 93, 132, 214, 326, 390, 474, 505, 796
MboII GAAGA 8 cut(s) 62, 65, 295, 484, 560, 575, 586, 964
MflI RGATCY 3 cut(s) 132, 505, 796
MhlI GDGCHC 2 cut(s) 386, 603
MluCI AATT 7 cut(s) 44, 343, 529, 733, 880, 896, 1120
MlyI GAGTC 1 cut(s) 470
MmeI TCCRAC 3 cut(s) 171, 273, 997
MroXI GAANNNNTTC 1 cut(s) 857
MspA1I CMGCKG 1 cut(s) 991
MspI CCGG 6 cut(s) 18, 156, 196, 432, 514, 713
MspR9I CCNGG 6 cut(s) 156, 196, 316, 514, 714, 1117
MvaI CCWGG 2 cut(s) 316, 1117
MvnI CGCG 1 cut(s) 67
MwoI GCNNNNNNNGC 7 cut(s) 70, 73, 186, 260, 490, 755, 1003
NciI CCSGG 4 cut(s) 156, 196, 514, 714
NcoI CCATGG 1 cut(s) 164
NdeI CATATG 1 cut(s) 419
NdeII GATC 8 cut(s) 93, 132, 214, 326, 390, 474, 505, 796
NlaIII CATG 3 cut(s) 168, 692, 1094
NlaIV GGNNCC 3 cut(s) 22, 718, 798
NmeAIII GCCGAG 3 cut(s) 154, 280, 364
NmuCI GTSAC 3 cut(s) 100, 292, 310
NsbI TGCGCA 1 cut(s) 371
NspV TTCGAA 1 cut(s) 561
PcsI WCGNNNNNNNCGW 2 cut(s) 456, 483
PdmI GAANNNNTTC 1 cut(s) 857
PfeI GAWTC 2 cut(s) 406, 985
PfoI TCCNGGA 2 cut(s) 712, 1115
PleI GAGTC 1 cut(s) 469
PpsI GAGTC 1 cut(s) 469
PshAI GACNNNNGTC 1 cut(s) 290
Psp124BI GAGCTC 1 cut(s) 386
Psp1406I AACGTT 1 cut(s) 375
Psp6I CCWGG 2 cut(s) 314, 1115
PspGI CCWGG 2 cut(s) 314, 1115
PspN4I GGNNCC 3 cut(s) 22, 718, 798
PspPI GGNCC 5 cut(s) 15, 161, 198, 318, 452
PstI CTGCAG 1 cut(s) 793
PstNI CAGNNNCTG 1 cut(s) 994
PsuI RGATCY 3 cut(s) 132, 505, 796
PvuII CAGCTG 1 cut(s) 991
RsaI GTAC 1 cut(s) 230
RsaNI GTAC 1 cut(s) 229
SacI GAGCTC 1 cut(s) 386
Sau3AI GATC 8 cut(s) 93, 132, 214, 326, 390, 474, 505, 796
Sau96I GGNCC 5 cut(s) 15, 161, 198, 318, 452
SchI GAGTC 1 cut(s) 470
ScrFI CCNGG 6 cut(s) 156, 196, 316, 514, 714, 1117
SduI GDGCHC 2 cut(s) 386, 603
SexAI ACCWGGT 1 cut(s) 314
SfaNI GCATC 2 cut(s) 354, 745
SfcI CTRYAG 1 cut(s) 789
SfuI TTCGAA 1 cut(s) 561
SinI GGWCC 2 cut(s) 161, 318
Sse9I AATT 7 cut(s) 44, 343, 529, 733, 880, 896, 1120
SsiI CCGC 8 cut(s) 65, 74, 159, 187, 482, 758, 770, 836
SstI GAGCTC 1 cut(s) 386
StyD4I CCNGG 6 cut(s) 154, 194, 314, 512, 712, 1115
StyI CCWWGG 2 cut(s) 164, 264
TaaI ACNGT 1 cut(s) 1042
TaiI ACGT 4 cut(s) 246, 294, 312, 378
TaqI TCGA 6 cut(s) 225, 459, 477, 546, 561, 808
TasI AATT 7 cut(s) 44, 343, 529, 733, 880, 896, 1120
TatI WGTACW 1 cut(s) 228
TauI GCSGC 3 cut(s) 67, 76, 484
TfiI GAWTC 2 cut(s) 406, 985
TscAI CASTG 4 cut(s) 37, 242, 708, 798
TseFI GTSAC 3 cut(s) 100, 292, 310
TseI GCWGC 7 cut(s) 367, 674, 746, 775, 788, 991, 994
Tsp45I GTSAC 3 cut(s) 100, 292, 310
TspDTI ATGAA 4 cut(s) 72, 659, 1088, 1163
TspGWI ACGGA 1 cut(s) 376
TspRI CASTG 4 cut(s) 37, 242, 708, 798
VneI GTGCAC 1 cut(s) 599
VpaK11BI GGWCC 2 cut(s) 161, 318
XapI RAATTY 4 cut(s) 529, 880, 896, 1120
XmiI GTMKAC 1 cut(s) 174
XmnI GAANNNNTTC 1 cut(s) 857
ZraI GACGTC 2 cut(s) 244, 292
Using CommOnly database (standard laboratory enzymes). Scanned on CDS sequence.