Rh2AG338500

Translation initiation factor eIF-2B subunit

Design Primers CRISPR Design

Basic Information

Type: gene
Biological Identity
rosa_samantha
Chr2A
OG0005877
Physical Location & Seq
Forward (+)
49034153 .. 49044133
9981 bp
Loading structure...
UTR
Exon/CDS
Intron
Rh2AG338500.1

Sequence Viewer

Length: 2139 bp
ATGGGGGCGCAGAAGAAAGGCGCGTCTAGGGCTTCAGAGGAACTGGCCCGTCTCCCACTCCAAGCCATCCTCTTAGCCGATAGTTTCACCACCAAATTCCGACCCATCACCCTTGAACGCCCCAAAGTGCTGTTGCCGTTAGTAAATGCCCCGATGATCAGTTACACATTAGCATGGCTTGAGTCTGCTGGCATTGAGGAGGTTTTTTTTTTCTGCTGTGCTCACTCCAAGCAAGTGATCAGTTATTTGGAGAATTCTGAGTGGTTTTCTCAACCAAACTTCAGGGTCACCACAATTGAGTTACATGATTCTGTTAGTGCCGGAGATGCTCTGCGCGTGATATATGAGCGCAATGTGATACATGGAGATTTTGTCCTAATCAGTGGGGATACCGTGAGCAACATGTCGCTTACTCAGGTGCTTCAAGAACATAAAGAGAGAAGAAAGAAAGATAGTAATGCCGCAATGACAATTGTTATTAAACGGTCAAAGCCTTCTCAAATCATTCACCAATGTCGACTGGGCACTGATGTGCTGTTTATGGCAATAGATCCCAACACAAAGCAGCTCTTATATTATGAGGACAAGGCAGACCATGCAAAAGGATCTATACATATAGATAAATTATTGCTTGCTGACAAATCTTCGATTTCTCTGCATAATGACAAACAGGATTGCTATATCGACATATGCTCTCCTGAAGTCCTCAGCCTTTTCACAGACAATTTTGACTATCACCATTTACGCTGTCATTTTTTAAAGGGATTGCTTGTTGATGATATTATGGGTTACAAAATATTCACACATGAAATTCACTCGACTTATGCGGCTAGGATTGACAACTTCAGAAGCTATGACACAATCAGTAGGGACATAATTCAGAGATGGGCTTACCCATTGGTGCCAGATGTTAAGTTTTTCAGAGATTCTTCAACGAAGCTGGAAAGACAGGGGATGTATCGAGTCTCAGAACTAGGGCAATCGCGCTCTGCACAGATTGGTCCTTTTACAGTCATTGGGAATGGCACCAAGATTGGGGACAACACCAAAATTTCAAATTCTGTTATTGGAGATGGGTGTTCTATTGGACCAAACGTTTCCATAGAAGGTTCCTATATATGGGATAATGTCACCATTGAAGATGGCTGCAAGATAAGGCATGCTATAGTATGTGATGGAGTGATAATGAAGTCTGGGGCAGTTTTGGAACCGGGTGTAGTTTTGTCTTTCAAGGTAGTGATAGGACAACAATTTATTGCTCCTTCATACTTAAAGGTATCTTTACTTCAACAACCAATAAAGCAAATTAGTGATGAGAAGCTAGAGTATGTTGACAATAGCACTGGTAATGCAGAAATTTCTTCTCTTACACATGCACTGGATATATCGAATGGGGAAATAGCGAACCAACAACAGGAGACACAGTGTTGGGCGACATCTGAGCTTGGTATTGGTGGGGCTGGTTATGTTTGGTCAATTTGTGAAGGAGGCCATGATGAAGGATGGAGGCATTCAGTTGCTCCCATTCCCACAGAGAAACTTGCTGAAGTTATACAAGCTGCAGAGGATGATCATGATGGCAGTCTCCCAACTTCAGATTCTGATTCTGATGAAGAGCCTAGAGACGACTCCTACTTTGAAAATGAGGTTGAAGCAACCTTCCTAAGAGCTGTGGATGAAAATATTAAAGTAGAGAATGTCGTTTTAGAAGTGAACTCACTGAGATTGTCATACAACAAGCAAACTGCAAACTGTGCTGGAGCTATATTTTATTCAATGATGAAATTGGCATTAGAATCTCCACATAATTCAGCAGGTGAATTGGTCCAAGCTACTATGAAGGTAATTACAAAATGGAAAGGACTGCTGAAGTATTACCTATCAGACATCGATGAAGAGATAGAAGTAATAATGAAATTTGAAGATATGTGTTTGGAGTCCGCTAAGGAGTTCTCCCCTGTGTTTGATAAGATACTCTATCAATTGTATGACCACACAATTATACGGGAAGAGGCTATTCTAAGGTGGGATGATGAAAAGAGAGATGCTGATGCATCAGATAGAATTTTTGTTAGGCAGGCTGAAGATTTTATCCAATGGTTGAGAGAGGCATCTGAAGAAGAGGATGAGGAAGAGTAG

Protein Analysis

712

Amino Acids

80.24

Weight (kDa)

4.89

Isoelectric Point (pI)

44.16

Instability Index
Protein Domains (Pfam)
Domain Name Pfam ID Position E-value Description
NTP_transferase PF00483 22 - 160 1.6e-11 Nucleotidyl transferase
LbH_EIF2B PF25084 316 - 418 4.3e-39 EIF2B subunit epsilon LbH domain
GMPPB_C PF25087 329 - 413 2.8e-10 GMPPB C-terminal domain
Hexapep PF00132 337 - 370 8.7e-08 Bacterial transferase hexapeptide (six repeats)
Hexapep_GlmU PF24894 338 - 401 7.6e-06 GlmU-like, C-terminal hexapeptide
W2 PF02020 633 - 711 4.1e-17 eIF4-gamma/eIF5/eIF2-epsilon
Hydrophobicity Profile (Kyte-Doolittle)
AI Structure Prediction Report
Calculating structure properties...

Genomic Context

Gene Family Tree

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Restriction Enzyme Sites

1 / 10
Enzyme Recognition Site Cut Count Positions (bp)
AarI CACCTGC 1 cut(s) 1805
Acc36I ACCTGC 1 cut(s) 1805
AccB1I GGYRCC 2 cut(s) 901, 1025
AccI GTMKAC 1 cut(s) 517
AccII CGCG 3 cut(s) 23, 336, 985
AciI CCGC 3 cut(s) 462, 827, 1941
AclI AACGTT 1 cut(s) 1095
AclWI GGATC 2 cut(s) 545, 613
AcsI RAATTY 8 cut(s) 95, 253, 810, 1050, 1057, 1356, 1916, 2064
AcuI CTGAAG 9 cut(s) 18, 265, 720, 829, 1566, 1578, 1889, 2103, 2136
AfiI CCNNNNNNNGG 3 cut(s) 1035, 1119, 1414
AflIII ACRYGT 1 cut(s) 402
AjuI GAANNNNNNNTTGG 2 cut(s) 1582, 1614
AleI CACNNNNGTG 1 cut(s) 530
AluBI AGCT 9 cut(s) 568, 852, 940, 1321, 1444, 1559, 1670, 1763, 1832
AluI AGCT 9 cut(s) 568, 852, 940, 1321, 1444, 1559, 1670, 1763, 1832
Alw21I GWGCWC 1 cut(s) 223
Alw26I GTCTC 5 cut(s) 56, 970, 1412, 1589, 1617
AlwI GGATC 2 cut(s) 545, 613
AlwNI CAGNNNCTG 1 cut(s) 1601
AoxI GGCC 2 cut(s) 45, 1489
ApeKI GCWGC 3 cut(s) 565, 1146, 1559
ApoI RAATTY 8 cut(s) 95, 253, 810, 1050, 1057, 1356, 1916, 2064
ArsI GACNNNNNNTTYG 2 cut(s) 1619, 1651
AspLEI GCGC 5 cut(s) 10, 23, 336, 351, 987
AspS9I GGNCC 4 cut(s) 46, 1001, 1088, 1825
AsuC2I CCSGG 1 cut(s) 1212
AsuHPI GGTGA 7 cut(s) 79, 100, 280, 500, 728, 1123, 1829
AvaII GGWCC 3 cut(s) 1001, 1088, 1825
BaeGI GKGCMC 1 cut(s) 527
BanI GGYRCC 2 cut(s) 901, 1025
BarI GAAGNNNNNNTAC 2 cut(s) 1269, 1301
Bbv12I GWGCWC 1 cut(s) 223
BbvCI CCTCAGC 1 cut(s) 707
BbvI GCAGC 3 cut(s) 577, 1133, 1546
BccI CCATC 8 cut(s) 74, 113, 879, 1067, 1136, 1169, 1497, 1571
BceAI ACGGC 1 cut(s) 121
BcgI CGANNNNNNTGC 2 cut(s) 637, 671
BciVI GTATCC 1 cut(s) 382
BclI TGATCA 3 cut(s) 156, 237, 1570
BcnI CCSGG 1 cut(s) 1212
BcoDI GTCTC 5 cut(s) 56, 970, 1412, 1589, 1617
BfaI CTAG 5 cut(s) 27, 831, 974, 1322, 1620
BfmI CTRYAG 2 cut(s) 1164, 1560
BfuAI ACCTGC 1 cut(s) 1805
BfuI GTATCC 1 cut(s) 382
BisI GCNGC 5 cut(s) 462, 566, 828, 1147, 1560
BlsI GCNGC 5 cut(s) 463, 567, 829, 1148, 1561
Bme1390I CCNGG 1 cut(s) 1212
Bme18I GGWCC 3 cut(s) 1001, 1088, 1825
BmgT120I GGNCC 4 cut(s) 46, 1001, 1088, 1825
BmiI GGNNCC 4 cut(s) 903, 1027, 1111, 1209
BmrFI CCNGG 1 cut(s) 1212
BmrI ACTGGG 1 cut(s) 530
BmsI GCATC 5 cut(s) 316, 2035, 2041, 2063, 2120
BmuI ACTGGG 1 cut(s) 530
BpmI CTGGAG 1 cut(s) 1779
Bpu10I CCTNAGC 2 cut(s) 707, 1944
BpuEI CTTGAG 1 cut(s) 200
BpuMI CCSGG 1 cut(s) 1212
Bsa29I ATCGAT 1 cut(s) 1890
BsaXI ACNNNNNCTCC 4 cut(s) 357, 387, 1409, 1439
Bsc4I CCNNNNNNNGG 3 cut(s) 1035, 1119, 1414
Bse1I ACTGG 4 cut(s) 48, 525, 1348, 1383
Bse3DI GCAATG 2 cut(s) 358, 471
BseCI ATCGAT 1 cut(s) 1890
BseGI GGATG 7 cut(s) 66, 960, 1508, 1573, 1681, 2035, 2131
BseLI CCNNNNNNNGG 3 cut(s) 1035, 1119, 1414
BseMI GCAATG 2 cut(s) 358, 471
BseMII CTCAG 6 cut(s) 249, 428, 721, 981, 1431, 1712
BseNI ACTGG 4 cut(s) 48, 525, 1348, 1383
BseRI GAGGAG 1 cut(s) 212
BseSI GKGCMC 1 cut(s) 527
BseXI GCAGC 3 cut(s) 577, 1133, 1546
BsgI GTGCAG 1 cut(s) 975
Bsh1236I CGCG 3 cut(s) 23, 336, 985
BshFI GGCC 2 cut(s) 47, 1491
BshNI GGYRCC 2 cut(s) 901, 1025
BshVI ATCGAT 1 cut(s) 1890
BsiHKAI GWGCWC 1 cut(s) 223
BsiSI CCGG 2 cut(s) 321, 1211
BslFI GGGAC 2 cut(s) 884, 1052
BslI CCNNNNNNNGG 3 cut(s) 1035, 1119, 1414
BsmAI GTCTC 5 cut(s) 56, 970, 1412, 1589, 1617
BsmBI CGTCTC 2 cut(s) 56, 1617
BsmFI GGGAC 2 cut(s) 884, 1052
BsmI GAATGC 1 cut(s) 1510
BsnI GGCC 2 cut(s) 47, 1491
Bsp1286I GDGCHC 2 cut(s) 223, 527
Bsp143I GATC 5 cut(s) 156, 237, 550, 605, 1570
BspACI CCGC 3 cut(s) 462, 827, 1941
BspANI GGCC 2 cut(s) 47, 1491
BspCNI CTCAG 6 cut(s) 250, 427, 720, 980, 1432, 1713
BspDI ATCGAT 1 cut(s) 1890
BspFNI CGCG 3 cut(s) 23, 336, 985
BspHI TCATGA 1 cut(s) 1573
BspLI GGNNCC 4 cut(s) 903, 1027, 1111, 1209
BspMAI CTGCAG 1 cut(s) 1564
BspMI ACCTGC 1 cut(s) 1805
BspPI GGATC 2 cut(s) 545, 613
BspQI GCTCTTC 1 cut(s) 1608
BspT107I GGYRCC 2 cut(s) 901, 1025
BsrDI GCAATG 2 cut(s) 358, 471
BsrI ACTGG 4 cut(s) 48, 525, 1348, 1383
BssMI GATC 5 cut(s) 156, 237, 550, 605, 1570
Bst4CI ACNGT 5 cut(s) 394, 486, 1012, 1425, 1754
Bst6I CTCTTC 5 cut(s) 1608, 1890, 2004, 2115, 2127
BstAPI GCANNNNNTGC 2 cut(s) 596, 1754
BstC8I GCNNGC 4 cut(s) 190, 633, 1161, 2079
BstEII GGTNACC 1 cut(s) 286
BstF5I GGATG 7 cut(s) 66, 960, 1508, 1573, 1681, 2035, 2131
BstFNI CGCG 3 cut(s) 23, 336, 985
BstHHI GCGC 5 cut(s) 10, 23, 336, 351, 987
BstKTI GATC 5 cut(s) 159, 240, 553, 608, 1573
BstMAI GTCTC 5 cut(s) 56, 970, 1412, 1589, 1617
BstMBI GATC 5 cut(s) 156, 237, 550, 605, 1570
BstMWI GCNNNNNNNGC 4 cut(s) 29, 326, 596, 1754
BstNSI RCATGY 3 cut(s) 406, 1163, 1376
BstPI GGTNACC 1 cut(s) 286
BstSCI CCNGG 1 cut(s) 1210
BstSFI CTRYAG 2 cut(s) 1164, 1560
BstSLI GKGCMC 1 cut(s) 527
BstUI CGCG 3 cut(s) 23, 336, 985
BstV1I GCAGC 3 cut(s) 577, 1133, 1546
BstX2I RGATCY 2 cut(s) 550, 605
BstYI RGATCY 2 cut(s) 550, 605
Bsu15I ATCGAT 1 cut(s) 1890
BsuI GTATCC 1 cut(s) 382
BsuRI GGCC 2 cut(s) 47, 1491
BsuTUI ATCGAT 1 cut(s) 1890
BtsCI GGATG 7 cut(s) 66, 960, 1508, 1573, 1681, 2035, 2131
BtsIMutI CAGTG 6 cut(s) 388, 525, 1341, 1376, 1430, 1718
BveI ACCTGC 1 cut(s) 1805
Cac8I GCNNGC 4 cut(s) 190, 633, 1161, 2079
CaiI CAGNNNCTG 1 cut(s) 1601
CciI TCATGA 1 cut(s) 1573
CfoI GCGC 5 cut(s) 10, 23, 336, 351, 987
Cfr13I GGNCC 4 cut(s) 46, 1001, 1088, 1825
ClaI ATCGAT 1 cut(s) 1890
CseI GACGC 1 cut(s) 12
DpnI GATC 5 cut(s) 158, 239, 552, 607, 1572
DpnII GATC 5 cut(s) 156, 237, 550, 605, 1570
DraI TTTAAA 1 cut(s) 759
Eam1104I CTCTTC 5 cut(s) 1608, 1890, 2004, 2115, 2127
EarI CTCTTC 5 cut(s) 1608, 1890, 2004, 2115, 2127
Eco47I GGWCC 3 cut(s) 1001, 1088, 1825
Eco57I CTGAAG 9 cut(s) 18, 265, 720, 829, 1566, 1578, 1889, 2103, 2136
Eco91I GGTNACC 1 cut(s) 286
EcoO65I GGTNACC 1 cut(s) 286
EcoRI GAATTC 1 cut(s) 253
EcoT22I ATGCAT 1 cut(s) 2056
Esp3I CGTCTC 2 cut(s) 56, 1617
FalI AAGNNNNNCTT 2 cut(s) 554, 586
FaqI GGGAC 2 cut(s) 884, 1052
FauNDI CATATG 1 cut(s) 689
FbaI TGATCA 3 cut(s) 156, 237, 1570
FblI GTMKAC 1 cut(s) 517
Fnu4HI GCNGC 5 cut(s) 462, 566, 828, 1147, 1560
FokI GGATG 6 cut(s) 53, 967, 1515, 1580, 1688, 2042
Fsp4HI GCNGC 5 cut(s) 462, 566, 828, 1147, 1560
FspBI CTAG 5 cut(s) 27, 831, 974, 1322, 1620
GlaI GCGC 5 cut(s) 9, 22, 335, 350, 986
GluI GCNGC 5 cut(s) 462, 566, 828, 1147, 1560
GsuI CTGGAG 1 cut(s) 1779
HaeIII GGCC 2 cut(s) 47, 1491
HapII CCGG 2 cut(s) 321, 1211
HgaI GACGC 1 cut(s) 12
HhaI GCGC 5 cut(s) 10, 23, 336, 351, 987
Hin6I GCGC 5 cut(s) 8, 21, 334, 349, 985
HinP1I GCGC 5 cut(s) 8, 21, 334, 349, 985
HincII GTYRAC 2 cut(s) 518, 1333
HindII GTYRAC 2 cut(s) 518, 1333
HinfI GANTC 9 cut(s) 182, 308, 926, 963, 1598, 1604, 1628, 1796, 1937
HpaII CCGG 2 cut(s) 321, 1211
HphI GGTGA 7 cut(s) 79, 100, 280, 500, 728, 1123, 1829
Hpy166II GTNNAC 3 cut(s) 518, 1333, 1714
Hpy188III TCNNGA 3 cut(s) 425, 698, 1574
Hpy8I GTNNAC 3 cut(s) 518, 1333, 1714
HpyAV CCTTC 7 cut(s) 504, 1100, 1272, 1478, 1493, 1669, 1834
HpyCH4III ACNGT 5 cut(s) 394, 486, 1012, 1425, 1754
HpyCH4IV ACGT 1 cut(s) 1095
HpyCH4V TGCA 9 cut(s) 599, 658, 992, 1149, 1352, 1376, 1562, 1748, 2054
HpyF10VI GCNNNNNNNGC 4 cut(s) 29, 326, 596, 1754
HpySE526I ACGT 1 cut(s) 1095
HspAI GCGC 5 cut(s) 8, 21, 334, 349, 985
Ksp22I TGATCA 3 cut(s) 156, 237, 1570
Kzo9I GATC 5 cut(s) 156, 237, 550, 605, 1570
LguI GCTCTTC 1 cut(s) 1608
LmnI GCTCC 3 cut(s) 1264, 1525, 1760
Lsp1109I GCAGC 3 cut(s) 577, 1133, 1546
LweI GCATC 5 cut(s) 316, 2035, 2041, 2063, 2120
MaeI CTAG 5 cut(s) 27, 831, 974, 1322, 1620
MaeII ACGT 1 cut(s) 1095
MaeIII GTNAC 5 cut(s) 161, 286, 300, 788, 1129
MalI GATC 5 cut(s) 158, 239, 552, 607, 1572
MboI GATC 5 cut(s) 156, 237, 550, 605, 1570
MfeI CAATTG 3 cut(s) 294, 471, 1982
MflI RGATCY 2 cut(s) 550, 605
MhlI GDGCHC 2 cut(s) 223, 527
MlyI GAGTC 4 cut(s) 191, 972, 1622, 1946
MmeI TCCRAC 1 cut(s) 124
Mph1103I ATGCAT 1 cut(s) 2056
MseI TTAA 5 cut(s) 480, 758, 912, 1271, 1686
MslI CAYNNNNRTG 2 cut(s) 172, 530
MspI CCGG 2 cut(s) 321, 1211
MspR9I CCNGG 1 cut(s) 1212
MunI CAATTG 3 cut(s) 294, 471, 1982
Mva1269I GAATGC 1 cut(s) 1510
MvnI CGCG 3 cut(s) 23, 336, 985
MwoI GCNNNNNNNGC 4 cut(s) 29, 326, 596, 1754
NciI CCSGG 1 cut(s) 1212
NdeI CATATG 1 cut(s) 689
NdeII GATC 5 cut(s) 156, 237, 550, 605, 1570
NlaIV GGNNCC 4 cut(s) 903, 1027, 1111, 1209
NmuCI GTSAC 2 cut(s) 286, 1129
NsiI ATGCAT 1 cut(s) 2056
NspI RCATGY 3 cut(s) 406, 1163, 1376
OliI CACNNNNGTG 1 cut(s) 530
PaeI GCATGC 1 cut(s) 1163
PagI TCATGA 1 cut(s) 1573
PaqCI CACCTGC 1 cut(s) 1805
PciI ACATGT 1 cut(s) 402
PciSI GCTCTTC 1 cut(s) 1608
PctI GAATGC 1 cut(s) 1510
PfeI GAWTC 5 cut(s) 308, 926, 1598, 1604, 1796
PkrI GCNGC 5 cut(s) 463, 567, 829, 1148, 1561
PleI GAGTC 4 cut(s) 190, 971, 1622, 1945
PpsI GAGTC 4 cut(s) 190, 971, 1622, 1945
PscI ACATGT 1 cut(s) 402
Psp1406I AACGTT 1 cut(s) 1095
PspEI GGTNACC 1 cut(s) 286
PspN4I GGNNCC 4 cut(s) 903, 1027, 1111, 1209
PspPI GGNCC 4 cut(s) 46, 1001, 1088, 1825
PstI CTGCAG 1 cut(s) 1564
PstNI CAGNNNCTG 1 cut(s) 1601
PsuI RGATCY 2 cut(s) 550, 605
RseI CAYNNNNRTG 2 cut(s) 172, 530
SalI GTCGAC 1 cut(s) 516
SapI GCTCTTC 1 cut(s) 1608
SaqAI TTAA 5 cut(s) 480, 758, 912, 1271, 1686
SatI GCNGC 5 cut(s) 462, 566, 828, 1147, 1560
Sau3AI GATC 5 cut(s) 156, 237, 550, 605, 1570
Sau96I GGNCC 4 cut(s) 46, 1001, 1088, 1825
SchI GAGTC 4 cut(s) 191, 972, 1622, 1946
ScrFI CCNGG 1 cut(s) 1212
SduI GDGCHC 2 cut(s) 223, 527
SfaNI GCATC 5 cut(s) 316, 2035, 2041, 2063, 2120
SfcI CTRYAG 2 cut(s) 1164, 1560
SinI GGWCC 3 cut(s) 1001, 1088, 1825
SmiMI CAYNNNNRTG 2 cut(s) 172, 530
SmlI CTYRAG 1 cut(s) 179
SmoI CTYRAG 1 cut(s) 179
SphI GCATGC 1 cut(s) 1163
SsiI CCGC 3 cut(s) 462, 827, 1941
SspI AATATT 2 cut(s) 798, 1684
SspMI CTAG 5 cut(s) 27, 831, 974, 1322, 1620
StyD4I CCNGG 1 cut(s) 1210
TaaI ACNGT 5 cut(s) 394, 486, 1012, 1425, 1754
TaiI ACGT 1 cut(s) 1098
TaqI TCGA 7 cut(s) 517, 647, 684, 818, 961, 1388, 1890
TauI GCSGC 2 cut(s) 464, 830
TfiI GAWTC 5 cut(s) 308, 926, 1598, 1604, 1796
Tru1I TTAA 5 cut(s) 480, 758, 912, 1271, 1686
Tru9I TTAA 5 cut(s) 480, 758, 912, 1271, 1686
TscAI CASTG 6 cut(s) 388, 532, 1348, 1383, 1430, 1725
TseFI GTSAC 2 cut(s) 286, 1129
TseI GCWGC 3 cut(s) 565, 1146, 1559
Tsp45I GTSAC 2 cut(s) 286, 1129
TspRI CASTG 6 cut(s) 388, 532, 1348, 1383, 1430, 1725
VpaK11BI GGWCC 3 cut(s) 1001, 1088, 1825
XapI RAATTY 8 cut(s) 95, 253, 810, 1050, 1057, 1356, 1916, 2064
XceI RCATGY 3 cut(s) 406, 1163, 1376
XmiI GTMKAC 1 cut(s) 517
XspI CTAG 5 cut(s) 27, 831, 974, 1322, 1620
Zsp2I ATGCAT 1 cut(s) 2056
Using CommOnly database (standard laboratory enzymes). Scanned on CDS sequence.