MD01G1237200.v1.1

Phosphate-induced protein 1 conserved region

Design Primers CRISPR Design

Basic Information

Type: gene
Biological Identity
malus_domestica
Chr01
OG0005922
Physical Location & Seq
Forward (+)
32426009 .. 32427076
1068 bp
Loading structure...
UTR
Exon/CDS
Intron
MD01G1237200.v1.1.491

Sequence Viewer

Length: 1068 bp
ATGCCAAACGCTGGTGGTGCTTATCCAATTCCAATTCCAGACGCTGGTGGTGCAAATCCAATTCCAATTCCGGACGCTGGTGGTGCAGCACCTGCTGCTGGCGGCACCGTCCCAGCACCTGGAACTGCTGGCGCTAAACCCGGTGGTGTTAATCCAACTCCAACTCCAACTCCAAACGCTGGTGGTGCAGCACATGCTACCGGCGGCACCGTCCCAGCTCCTGGAGTTGGCGGACCCATCACATACCACGGAGGGCCTCTGATGACCGGCAACATTAACCTTTCCATCCTATTTTACGGTGAGTTCAAATCCGAACAGAGGGAAGTGATTCGGACTTTCCTGAGGTCCCTCGAGAACACGGAGAACGATCACGTCGCGGCGGTGCACAGGTGGTGGGACATCGTCGAGAGCTACCAATTGTTCGCTAGTCCAACCCCCGGTAAAGGTACCCCACCCAAACTCCGCATCAAGGTGGGGACCCAGCAGAGCGACGACAAGTACTCGATTGGAAAAGTGATAACCTATGAGTTCATCAAGAGACTGTTAAAGAAGGCCGACTCCGGAAAGCCCAACACTATTACCGTGATCTTTACCGGGAGCCAAGTGACGGTGCACAGCTTGTGCCGTGGCAGATGCTACGAGCATGGCTTGGTTGAGAACAGACCCTACATCGTCGTCGGCAACCCCGAAATCTTGTGCTCCGGGAGTTGCGCTTGGCCATTCCAGAGGCTGGACTACGGTACTCCTAATCAAGTGCCGGTGAAGCCCCCCAACGGCGACATGGGCATTGACGCCATGGTCGTCCACTTCGCCTCCGCCTTGGCTTCCGCCGTCACCAACCCATTCAACACTGGATTCTTCAAGCCGGGACCCAAGAGCGACCCCATCGAAGCCGTCACAGCCTGCGAAAACATCTTCGGAAGCGGCGCCGTCCCAGGCCAGACCGGCAAGGTTGAGGTCGACCCCGCTACCGGTGCCTCCTACAATGCCATCGGAGAGAATGGTATGAAGTTCATGCTCCCGGCCGTCTGGGACCCACGAACTTCTTCCTGCTGGACCTCCATGTAA
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
Phi_1

Protein Analysis

356

Amino Acids

37.04

Weight (kDa)

7.04

Isoelectric Point (pI)

27.6

Instability Index
Protein Domains (Pfam)
Domain Name Pfam ID Position E-value Description
Phi_1 PF04674 80 - 353 1.3e-91 Phosphate-induced protein 1 conserved region
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) 100
AasI GACNNNNNNGTC 1 cut(s) 797
Acc36I ACCTGC 1 cut(s) 100
Acc65I GGTACC 1 cut(s) 446
AccB1I GGYRCC 5 cut(s) 104, 206, 446, 926, 974
AccB7I CCANNNNNTGG 6 cut(s) 11, 44, 119, 179, 221, 730
AccI GTMKAC 1 cut(s) 960
AccII CGCG 1 cut(s) 377
AccIII TCCGGA 2 cut(s) 70, 560
AcoI YGGCCR 2 cut(s) 716, 1023
AcyI GRCGYC 2 cut(s) 792, 927
AfaI GTAC 3 cut(s) 448, 500, 742
AgeI ACCGGT 1 cut(s) 971
AgsI TTSAA 3 cut(s) 307, 847, 862
AjiI CACGTC 1 cut(s) 373
AjnI CCWGG 3 cut(s) 118, 220, 934
AluBI AGCT 3 cut(s) 218, 411, 618
AluI AGCT 3 cut(s) 218, 411, 618
Alw21I GWGCWC 3 cut(s) 387, 615, 701
Alw26I GTCTC 1 cut(s) 532
Alw44I GTGCAC 2 cut(s) 383, 611
AlwNI CAGNNNCTG 5 cut(s) 44, 92, 119, 221, 730
Ama87I CYCGRG 1 cut(s) 350
Aor13HI TCCGGA 2 cut(s) 70, 560
AoxI GGCC 5 cut(s) 254, 552, 716, 937, 1023
ApaLI GTGCAC 2 cut(s) 383, 611
ApeKI GCWGC 3 cut(s) 86, 95, 188
AsiGI ACCGGT 1 cut(s) 971
Asp700I GAANNNNTTC 2 cut(s) 327, 1045
Asp718I GGTACC 1 cut(s) 446
AspLEI GCGC 3 cut(s) 134, 713, 929
AspS9I GGNCC 7 cut(s) 233, 254, 345, 477, 869, 1033, 1056
AsuC2I CCSGG 6 cut(s) 141, 438, 595, 703, 867, 1022
AsuHPI GGTGA 3 cut(s) 311, 772, 826
AvaI CYCGRG 1 cut(s) 350
AvaII GGWCC 6 cut(s) 233, 345, 477, 869, 1033, 1056
AxyI CCTNAGG 1 cut(s) 341
BaeGI GKGCMC 2 cut(s) 387, 615
BalI TGGCCA 1 cut(s) 718
BanI GGYRCC 5 cut(s) 104, 206, 446, 926, 974
Bbv12I GWGCWC 3 cut(s) 387, 615, 701
BbvI GCAGC 3 cut(s) 82, 98, 200
BccI CCATC 4 cut(s) 245, 293, 893, 998
BceAI ACGGC 6 cut(s) 609, 790, 815, 878, 914, 1010
BciT130I CCWGG 3 cut(s) 120, 222, 936
BcnI CCSGG 6 cut(s) 141, 438, 595, 703, 867, 1022
BcoDI GTCTC 1 cut(s) 532
BfaI CTAG 1 cut(s) 426
BfoI RGCGCY 2 cut(s) 135, 930
BfuAI ACCTGC 1 cut(s) 100
BglI GCCNNNNNGGC 1 cut(s) 945
BisI GCNGC 7 cut(s) 87, 96, 103, 189, 205, 378, 925
BlsI GCNGC 7 cut(s) 88, 97, 104, 190, 206, 379, 926
BmcAI AGTACT 1 cut(s) 500
Bme1390I CCNGG 9 cut(s) 120, 141, 222, 438, 595, 703, 867, 936, 1022
Bme18I GGWCC 6 cut(s) 233, 345, 477, 869, 1033, 1056
BmeT110I CYCGRG 1 cut(s) 350
BmgBI CACGTC 1 cut(s) 373
BmgT120I GGNCC 7 cut(s) 233, 254, 345, 477, 869, 1033, 1056
BmrFI CCNGG 9 cut(s) 120, 141, 222, 438, 595, 703, 867, 936, 1022
BmsI GCATC 2 cut(s) 474, 623
BpmI CTGGAG 1 cut(s) 243
BpuMI CCSGG 6 cut(s) 141, 438, 595, 703, 867, 1022
BsaHI GRCGYC 2 cut(s) 792, 927
BsaJI CCNNGG 6 cut(s) 247, 436, 625, 795, 819, 934
BsaWI WCCGGW 3 cut(s) 70, 560, 971
BsaXI ACNNNNNCTCC 8 cut(s) 148, 154, 178, 184, 444, 474, 797, 827
Bse118I RCCGGY 5 cut(s) 200, 266, 757, 944, 971
Bse1I ACTGG 1 cut(s) 856
Bse21I CCTNAGG 1 cut(s) 341
BseAI TCCGGA 2 cut(s) 70, 560
BseBI CCWGG 3 cut(s) 120, 222, 936
BseDI CCNNGG 6 cut(s) 247, 436, 625, 795, 819, 934
BseGI GGATG 1 cut(s) 285
BseMII CTCAG 1 cut(s) 332
BseNI ACTGG 1 cut(s) 856
BseSI GKGCMC 2 cut(s) 387, 615
BseX3I CGGCCG 1 cut(s) 1023
BseXI GCAGC 3 cut(s) 82, 98, 200
BseYI CCCAGC 3 cut(s) 112, 214, 480
BsgI GTGCAG 2 cut(s) 105, 207
Bsh1236I CGCG 1 cut(s) 377
Bsh1285I CGRYCG 1 cut(s) 1026
BshFI GGCC 5 cut(s) 256, 554, 718, 939, 1025
BshNI GGYRCC 5 cut(s) 104, 206, 446, 926, 974
BshTI ACCGGT 1 cut(s) 971
BsiEI CGRYCG 1 cut(s) 1026
BsiHKAI GWGCWC 3 cut(s) 387, 615, 701
BsiHKCI CYCGRG 1 cut(s) 350
BslFI GGGAC 8 cut(s) 95, 197, 331, 410, 490, 882, 917, 1046
BsmAI GTCTC 1 cut(s) 532
BsmFI GGGAC 8 cut(s) 95, 197, 331, 410, 490, 882, 917, 1046
BsnI GGCC 5 cut(s) 256, 554, 718, 939, 1025
BsoBI CYCGRG 1 cut(s) 350
Bsp1286I GDGCHC 3 cut(s) 387, 615, 701
Bsp13I TCCGGA 2 cut(s) 70, 560
Bsp143I GATC 2 cut(s) 367, 585
Bsp19I CCATGG 1 cut(s) 795
BspANI GGCC 5 cut(s) 256, 554, 718, 939, 1025
BspCNI CTCAG 1 cut(s) 333
BspEI TCCGGA 2 cut(s) 70, 560
BspFNI CGCG 1 cut(s) 377
BspMI ACCTGC 1 cut(s) 100
BspT107I GGYRCC 5 cut(s) 104, 206, 446, 926, 974
BsrFI RCCGGY 5 cut(s) 200, 266, 757, 944, 971
BsrI ACTGG 1 cut(s) 856
BssAI RCCGGY 5 cut(s) 200, 266, 757, 944, 971
BssECI CCNNGG 6 cut(s) 247, 436, 625, 795, 819, 934
BssMI GATC 2 cut(s) 367, 585
BssNI GRCGYC 2 cut(s) 792, 927
BssT1I CCWWGG 2 cut(s) 795, 819
Bst2UI CCWGG 3 cut(s) 120, 222, 936
Bst4CI ACNGT 7 cut(s) 109, 211, 299, 543, 583, 610, 740
BstACI GRCGYC 2 cut(s) 792, 927
BstAPI GCANNNNNTGC 3 cut(s) 92, 95, 194
BstC8I GCNNGC 3 cut(s) 100, 130, 904
BstDEI CTNAG 1 cut(s) 341
BstDSI CCRYGG 3 cut(s) 247, 625, 795
BstF5I GGATG 1 cut(s) 285
BstFNI CGCG 1 cut(s) 377
BstH2I RGCGCY 2 cut(s) 135, 930
BstHHI GCGC 3 cut(s) 134, 713, 929
BstKTI GATC 2 cut(s) 370, 588
BstMAI GTCTC 1 cut(s) 532
BstMBI GATC 2 cut(s) 367, 585
BstMCI CGRYCG 1 cut(s) 1026
BstNI CCWGG 3 cut(s) 120, 222, 936
BstNSI RCATGY 1 cut(s) 197
BstSCI CCNGG 9 cut(s) 118, 139, 220, 436, 593, 701, 865, 934, 1020
BstSLI GKGCMC 2 cut(s) 387, 615
BstUI CGCG 1 cut(s) 377
BstV1I GCAGC 3 cut(s) 82, 98, 200
BstZI CGGCCG 1 cut(s) 1023
Bsu36I CCTNAGG 1 cut(s) 341
BsuRI GGCC 5 cut(s) 256, 554, 718, 939, 1025
BtgI CCRYGG 3 cut(s) 247, 625, 795
BtrI CACGTC 1 cut(s) 373
BtsCI GGATG 1 cut(s) 285
BtsIMutI CAGTG 1 cut(s) 849
BveI ACCTGC 1 cut(s) 100
Cac8I GCNNGC 3 cut(s) 100, 130, 904
CaiI CAGNNNCTG 5 cut(s) 44, 92, 119, 221, 730
CfoI GCGC 3 cut(s) 134, 713, 929
Cfr10I RCCGGY 5 cut(s) 200, 266, 757, 944, 971
Cfr13I GGNCC 7 cut(s) 233, 254, 345, 477, 869, 1033, 1056
CseI GACGC 3 cut(s) 50, 83, 800
Csp6I GTAC 3 cut(s) 447, 499, 741
CspAI ACCGGT 1 cut(s) 971
CviAII CATG 6 cut(s) 194, 644, 781, 796, 1015, 1063
CviQI GTAC 3 cut(s) 447, 499, 741
DdeI CTNAG 1 cut(s) 341
DinI GGCGCC 1 cut(s) 928
DpnI GATC 2 cut(s) 369, 587
DpnII GATC 2 cut(s) 367, 585
DrdI GACNNNNNNGTC 1 cut(s) 797
DseDI GACNNNNNNGTC 1 cut(s) 797
EaeI YGGCCR 2 cut(s) 716, 1023
EagI CGGCCG 1 cut(s) 1023
EciI GGCGGA 3 cut(s) 246, 805, 817
EclXI CGGCCG 1 cut(s) 1023
Eco130I CCWWGG 2 cut(s) 795, 819
Eco47I GGWCC 6 cut(s) 233, 345, 477, 869, 1033, 1056
Eco52I CGGCCG 1 cut(s) 1023
Eco81I CCTNAGG 1 cut(s) 341
Eco88I CYCGRG 1 cut(s) 350
EcoO109I RGGNCCY 5 cut(s) 254, 345, 477, 869, 1033
EcoRII CCWGG 3 cut(s) 118, 220, 934
EcoT14I CCWWGG 2 cut(s) 795, 819
EgeI GGCGCC 1 cut(s) 928
EheI GGCGCC 1 cut(s) 928
ErhI CCWWGG 2 cut(s) 795, 819
FaeI CATG 6 cut(s) 197, 647, 784, 799, 1018, 1066
FaiI YATR 9 cut(s) 195, 244, 525, 645, 782, 797, 1007, 1016, 1064
FaqI GGGAC 8 cut(s) 95, 197, 331, 410, 490, 882, 917, 1046
FatI CATG 6 cut(s) 193, 643, 780, 795, 1014, 1062
FauI CCCGC 1 cut(s) 973
FblI GTMKAC 1 cut(s) 960
Fnu4HI GCNGC 7 cut(s) 87, 96, 103, 189, 205, 378, 925
FokI GGATG 1 cut(s) 272
Fsp4HI GCNGC 7 cut(s) 87, 96, 103, 189, 205, 378, 925
FspBI CTAG 1 cut(s) 426
GlaI GCGC 3 cut(s) 133, 712, 928
GluI GCNGC 7 cut(s) 87, 96, 103, 189, 205, 378, 925
GsaI CCCAGC 3 cut(s) 116, 218, 484
GsuI CTGGAG 1 cut(s) 243
HaeII RGCGCY 2 cut(s) 135, 930
HaeIII GGCC 5 cut(s) 256, 554, 718, 939, 1025
HgaI GACGC 3 cut(s) 50, 83, 800
HhaI GCGC 3 cut(s) 134, 713, 929
Hin1I GRCGYC 2 cut(s) 792, 927
Hin1II CATG 6 cut(s) 197, 647, 784, 799, 1018, 1066
Hin6I GCGC 3 cut(s) 132, 711, 927
HinP1I GCGC 3 cut(s) 132, 711, 927
HincII GTYRAC 1 cut(s) 961
HindII GTYRAC 1 cut(s) 961
HinfI GANTC 3 cut(s) 328, 557, 855
HphI GGTGA 3 cut(s) 311, 772, 826
Hpy166II GTNNAC 4 cut(s) 385, 613, 805, 961
Hpy188I TCNGA 5 cut(s) 261, 313, 333, 920, 995
Hpy188III TCNNGA 8 cut(s) 38, 71, 340, 352, 406, 535, 561, 724
Hpy8I GTNNAC 4 cut(s) 385, 613, 805, 961
Hpy99I CGWCG 5 cut(s) 377, 407, 494, 677, 680
HpyAV CCTTC 1 cut(s) 544
HpyCH4III ACNGT 7 cut(s) 109, 211, 299, 543, 583, 610, 740
HpyCH4IV ACGT 1 cut(s) 372
HpyCH4V TGCA 5 cut(s) 53, 86, 188, 385, 613
HpyF3I CTNAG 1 cut(s) 341
HpySE526I ACGT 1 cut(s) 372
Hsp92I GRCGYC 2 cut(s) 792, 927
Hsp92II CATG 6 cut(s) 197, 647, 784, 799, 1018, 1066
HspAI GCGC 3 cut(s) 132, 711, 927
KasI GGCGCC 1 cut(s) 926
KflI GGGWCCC 3 cut(s) 477, 869, 1033
Kpn2I TCCGGA 2 cut(s) 70, 560
KpnI GGTACC 1 cut(s) 450
Kzo9I GATC 2 cut(s) 367, 585
LmnI GCTCC 4 cut(s) 223, 597, 704, 1023
Lsp1109I GCAGC 3 cut(s) 82, 98, 200
LweI GCATC 2 cut(s) 474, 623
MaeI CTAG 1 cut(s) 426
MaeII ACGT 1 cut(s) 372
MaeIII GTNAC 3 cut(s) 604, 832, 895
MalI GATC 2 cut(s) 369, 587
MboI GATC 2 cut(s) 367, 585
MboII GAAGA 3 cut(s) 850, 907, 1038
MfeI CAATTG 1 cut(s) 416
MhlI GDGCHC 3 cut(s) 387, 615, 701
MlsI TGGCCA 1 cut(s) 718
MluCI AATT 5 cut(s) 27, 33, 60, 66, 416
MluNI TGGCCA 1 cut(s) 718
Mly113I GGCGCC 1 cut(s) 927
MlyI GAGTC 1 cut(s) 551
MmeI TCCRAC 4 cut(s) 179, 185, 191, 455
MnlI CCTC 9 cut(s) 245, 267, 312, 336, 359, 720, 823, 949, 988
Mox20I TGGCCA 1 cut(s) 718
MroI TCCGGA 2 cut(s) 70, 560
MroXI GAANNNNTTC 2 cut(s) 327, 1045
MscI TGGCCA 1 cut(s) 718
MseI TTAA 3 cut(s) 150, 276, 545
MslI CAYNNNNRTG 1 cut(s) 470
Msp20I TGGCCA 1 cut(s) 718
MspR9I CCNGG 9 cut(s) 120, 141, 222, 438, 595, 703, 867, 936, 1022
MunI CAATTG 1 cut(s) 416
MvaI CCWGG 3 cut(s) 120, 222, 936
MvnI CGCG 1 cut(s) 377
NarI GGCGCC 1 cut(s) 927
NciI CCSGG 6 cut(s) 141, 438, 595, 703, 867, 1022
NcoI CCATGG 1 cut(s) 795
NdeII GATC 2 cut(s) 367, 585
NlaIII CATG 6 cut(s) 197, 647, 784, 799, 1018, 1066
NmuCI GTSAC 3 cut(s) 604, 832, 895
NspI RCATGY 1 cut(s) 197
PaeR7I CTCGAG 1 cut(s) 350
PaqCI CACCTGC 1 cut(s) 100
PcsI WCGNNNNNNNCGW 2 cut(s) 684, 798
PdmI GAANNNNTTC 2 cut(s) 327, 1045
PfeI GAWTC 2 cut(s) 328, 855
PflFI GACNNNGTC 1 cut(s) 401
PflMI CCANNNNNTGG 6 cut(s) 11, 44, 119, 179, 221, 730
PfoI TCCNGGA 2 cut(s) 220, 701
PinAI ACCGGT 1 cut(s) 971
PkrI GCNGC 7 cut(s) 88, 97, 104, 190, 206, 379, 926
PleI GAGTC 1 cut(s) 551
PluTI GGCGCC 1 cut(s) 930
PpsI GAGTC 1 cut(s) 551
PpuMI RGGWCCY 4 cut(s) 345, 477, 869, 1033
Psp5II RGGWCCY 4 cut(s) 345, 477, 869, 1033
Psp6I CCWGG 3 cut(s) 118, 220, 934
PspFI CCCAGC 3 cut(s) 112, 214, 480
PspGI CCWGG 3 cut(s) 118, 220, 934
PspPI GGNCC 7 cut(s) 233, 254, 345, 477, 869, 1033, 1056
PspPPI RGGWCCY 4 cut(s) 345, 477, 869, 1033
PsrI GAACNNNNNNTAC 2 cut(s) 650, 682
PstNI CAGNNNCTG 5 cut(s) 44, 92, 119, 221, 730
PsyI GACNNNGTC 1 cut(s) 401
RsaI GTAC 3 cut(s) 448, 500, 742
RsaNI GTAC 3 cut(s) 447, 499, 741
RseI CAYNNNNRTG 1 cut(s) 470
SalI GTCGAC 1 cut(s) 959
SaqAI TTAA 3 cut(s) 150, 276, 545
SatI GCNGC 7 cut(s) 87, 96, 103, 189, 205, 378, 925
Sau3AI GATC 2 cut(s) 367, 585
Sau96I GGNCC 7 cut(s) 233, 254, 345, 477, 869, 1033, 1056
ScaI AGTACT 1 cut(s) 500
SchI GAGTC 1 cut(s) 551
ScrFI CCNGG 9 cut(s) 120, 141, 222, 438, 595, 703, 867, 936, 1022
SduI GDGCHC 3 cut(s) 387, 615, 701
SfaNI GCATC 2 cut(s) 474, 623
SfoI GGCGCC 1 cut(s) 928
Sfr274I CTCGAG 1 cut(s) 350
SinI GGWCC 6 cut(s) 233, 345, 477, 869, 1033, 1056
SlaI CTCGAG 1 cut(s) 350
SmiMI CAYNNNNRTG 1 cut(s) 470
SmlI CTYRAG 1 cut(s) 350
SmoI CTYRAG 1 cut(s) 350
Sse9I AATT 5 cut(s) 27, 33, 60, 66, 416
SspDI GGCGCC 1 cut(s) 926
SspMI CTAG 1 cut(s) 426
StyD4I CCNGG 9 cut(s) 118, 139, 220, 436, 593, 701, 865, 934, 1020
StyI CCWWGG 2 cut(s) 795, 819
TaaI ACNGT 7 cut(s) 109, 211, 299, 543, 583, 610, 740
TaiI ACGT 1 cut(s) 375
TaqI TCGA 5 cut(s) 351, 405, 503, 888, 960
TasI AATT 5 cut(s) 27, 33, 60, 66, 416
TatI WGTACW 1 cut(s) 498
TauI GCSGC 4 cut(s) 105, 207, 380, 927
TfiI GAWTC 2 cut(s) 328, 855
Tru1I TTAA 3 cut(s) 150, 276, 545
Tru9I TTAA 3 cut(s) 150, 276, 545
TscAI CASTG 1 cut(s) 856
TseFI GTSAC 3 cut(s) 604, 832, 895
TseI GCWGC 3 cut(s) 86, 95, 188
Tsp45I GTSAC 3 cut(s) 604, 832, 895
TspDTI ATGAA 3 cut(s) 520, 1003, 1022
TspGWI ACGGA 2 cut(s) 264, 374
TspRI CASTG 1 cut(s) 856
Tth111I GACNNNGTC 1 cut(s) 401
Van91I CCANNNNNTGG 6 cut(s) 11, 44, 119, 179, 221, 730
VneI GTGCAC 2 cut(s) 383, 611
VpaK11BI GGWCC 6 cut(s) 233, 345, 477, 869, 1033, 1056
XceI RCATGY 1 cut(s) 197
XcmI CCANNNNNNNNNTGG 1 cut(s) 778
XhoI CTCGAG 1 cut(s) 350
XmiI GTMKAC 1 cut(s) 960
XmnI GAANNNNTTC 2 cut(s) 327, 1045
XspI CTAG 1 cut(s) 426
ZrmI AGTACT 1 cut(s) 500
Using CommOnly database (standard laboratory enzymes). Scanned on CDS sequence.