RLG00000028703

GRAM domain-containing protein

Basic Information

Type: gene
Biological Identity
rosa_laevigata
Chr6
Physical Location & Seq
Reverse (-)
26724629 .. 26732989
8361 bp
Loading structure...
UTR
Exon/CDS
Intron
RLM00000028703

Sequence Viewer

Length: 1938 bp
ATGGCCGTGGCCTCCGCCTCCCCTGACAGGATCCAAGTCACGCCGCAGCAGCAGCAGCACCCGGTCATCGAGCCCTCGCCGTCTAGATTCTCCACCGACTCCGACGCCGCCTCGTCTTCACCGCTCGTCTCCTGCTCCGCCGACACGCCTGATCGCTCCGATCCTTCCTTTGCTTCTCCCAACTCCGATCAGTCTGCTGCAAGATTGAGGAGTGAAGAGTATCGGCAGTTGTTTCGGCTTCCAGCTGAAGAAGCCCTCATCGAAGATTTCAATTGTGCATTCCAGGAGAATATACTTATTCAGGGTCATATGTACCTGTTTGTTCATTATATTTGCTTTTACTCAAATATATTTGGATATGAGACAAAGAAAATAATCCCATTCCAGGAAGTCACATGCGTAAGAAGGGCGAAGACAGCAGGGATCTTTCCTAATGCAATAGAAATTTTTGTTGGAGACAAGAAGTACTTCTTTGCGTCCTTTCTGTCCCGTGATGAAGCTTACAGAATCATAAATGATGGTTGGGCACAATATGTTGATGGTGCCAAAGTAACTACAGAACAACAGGAATCAATCACTGAGACTAAAAACCAAGAGAATGGAGTTGTAATTGAAAAAGTGGAGAGCTCGAATTGCTTAGTTAATGGATTGGATTCTACTGAAAGGAACACTAATACTCAAATTTCGAAAGATTCTGAGCTCCCACCCAAAGTTGAAGACGATTCTGTGTTGACATCGCGTCCAGAGCCCAAAGGTGTTATCAAAGAGGATATAGAACCAGTTCTGAATGGTGAACCTTCATCTTCAAGAGATACGTTGATATTCAAGGAAGAAGATTCTGATGCCCCAAACATTCCAGAATTTTATACAAAGGTTGCGGAGTCGCAGTTTCCGATAAAGGTTGAAGATTTCTTTAGTTTCTTTTACTCTGATGATGTGTTCATTGAATCATTCCATAACAGTTGTGGAGATAAAGAGTTCAGCTGCACTTCATGGCAGCCTCATGAAAAGTTTGGACATACCCGCGATGTTTCATTTCAGCATCCAATTAAAATCTACTTTGGTGCAAAATTTGGTAGCTGTAAGGAGCTCCAGAAATTTCGAGTACACAGAAACAGTCATTTAGTTATCGAGACATCACAAGAGATCAACGATGTGCCTTATGGGGATTATTTCCGGGTTGAGGGGCTTTGGAATGTGGAGAGAGATACTGATGGATCAAAGGAATGCTGCAATTTGAAGATTTACTTGAATGTGGCATTTTCAAAAAGAACTGTTTGGAAAGGAAAGATAGTGCAGTCTACGTTGGAAGAATGTCGAGAGGTTTTTGCTACCTGGATAAACATGGCACATGAATTGTTGAAGACGAAGAACCTTGAGAAACAAGAAGAGAGCATTCCTACGGTTAGCATGGTTCAGATATGTGAAGTTCATCCTACAGAGGAAGCAGAGACGGTTCAGTCTTCTGAAAAGTTTTACGAGCCTAGTCATCATACTAGGATACAACAGATATCTGCATCCATAAACCCTAAGCACAAAGTTAGTCTATTTCAAGAAAGTCCGAGTGATACTTACTCTCTTACCTCATGGTTGAGAGACTATATGATGAAATTCGGTTCATCTTTGAGGAATCAAAGCCATCTTCCATTGATCATGGTTATCATTGTTGCCGTGGTTTTCATAATGCAGTTGAGCATACTTGCACTACTAGCTAGACCCCAACACATCCATGTGCACTCTCCTGTGGACTATACAAGTGTCGTGGGCAGTGGTGTTGGCGGGAGATCTTCTGAAGTTATTACTTGGCTGGAGAAGCGAGTTCACAACCTTAAAGACGAGATGAACTTGGTTGAAGCTCGGCTTGAGAGCATGCGACATGAACATGCTCTCCTGAAAGCTCAACTTGAAGATTTCAAGCATCTTATTAAGCGTAAATAA

Protein Analysis

646

Amino Acids

73.61

Weight (kDa)

5.43

Isoelectric Point (pI)

59.3

Instability Index
Protein Domains (Pfam)
Domain Name Pfam ID Position E-value Description
GRAM PF02893 74 - 177 1.3e-24 GRAM domain
VASt PF16016 292 - 437 4.3e-35 VAD1 Analog of StAR-related lipid transfer domain
Hydrophobicity Profile (Kyte-Doolittle)
AI Structure Prediction Report
Calculating structure properties...

Genomic Context

Gene Family Tree

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Publication-ready

Restriction Enzyme Sites

1 / 10
Enzyme Recognition Site Cut Count Positions (bp)
AasI GACNNNNNNGTC 1 cut(s) 1459
AccB1I GGYRCC 1 cut(s) 542
AccBSI CCGCTC 1 cut(s) 124
AccI GTMKAC 1 cut(s) 1301
AccII CGCG 2 cut(s) 739, 1026
AciI CCGC 8 cut(s) 15, 44, 108, 122, 138, 878, 1024, 1779
AclWI GGATC 5 cut(s) 25, 38, 155, 431, 1225
AcoI YGGCCR 1 cut(s) 3
AcsI RAATTY 6 cut(s) 444, 681, 860, 1070, 1097, 1610
AcuI CTGAAG 2 cut(s) 267, 1812
AcyI GRCGYC 1 cut(s) 105
AfaI GTAC 3 cut(s) 314, 467, 1107
AfiI CCNNNNNNNGG 3 cut(s) 27, 385, 1183
AhdI GACNNNNNGTC 1 cut(s) 738
AjnI CCWGG 3 cut(s) 282, 384, 1334
AjuI GAANNNNNNNTTGG 2 cut(s) 435, 467
AloI GAACNNNNNNTCC 2 cut(s) 1872, 1904
Alw21I GWGCWC 4 cut(s) 629, 702, 1092, 1737
Alw26I GTCTC 7 cut(s) 133, 356, 450, 575, 1127, 1445, 1590
Alw44I GTGCAC 1 cut(s) 1733
AlwI GGATC 5 cut(s) 25, 38, 155, 431, 1225
AoxI GGCC 2 cut(s) 3, 9
ApaLI GTGCAC 1 cut(s) 1733
ApeKI GCWGC 8 cut(s) 46, 49, 52, 55, 197, 984, 997, 1230
ApoI RAATTY 6 cut(s) 444, 681, 860, 1070, 1097, 1610
Asp700I GAANNNNTTC 2 cut(s) 467, 780
AsuC2I CCSGG 2 cut(s) 62, 1178
AsuHPI GGTGA 2 cut(s) 111, 803
AsuII TTCGAA 1 cut(s) 686
BaeGI GKGCMC 2 cut(s) 529, 1737
BamHI GGATCC 1 cut(s) 30
BanI GGYRCC 1 cut(s) 542
BanII GRGCYC 5 cut(s) 75, 629, 702, 750, 1092
BbsI GAAGAC 5 cut(s) 108, 419, 723, 1370, 1455
Bbv12I GWGCWC 4 cut(s) 629, 702, 1092, 1737
BbvI GCAGC 8 cut(s) 58, 61, 64, 67, 184, 971, 1009, 1217
BccI CCATC 4 cut(s) 512, 533, 1208, 1647
BceAI ACGGC 2 cut(s) 64, 1655
BcgI CGANNNNNNTGC 4 cut(s) 176, 210, 215, 249
BciT130I CCWGG 3 cut(s) 284, 386, 1336
BciVI GTATCC 1 cut(s) 1494
BclI TGATCA 1 cut(s) 1650
BcnI CCSGG 2 cut(s) 62, 1178
BcoDI GTCTC 7 cut(s) 133, 356, 450, 575, 1127, 1445, 1590
BfaI CTAG 5 cut(s) 84, 1485, 1497, 1709, 1713
BfmI CTRYAG 2 cut(s) 555, 1437
BfuI GTATCC 1 cut(s) 1494
BglII AGATCT 1 cut(s) 1784
BmcAI AGTACT 1 cut(s) 467
Bme1390I CCNGG 5 cut(s) 62, 284, 386, 1178, 1336
BmeRI GACNNNNNGTC 1 cut(s) 738
BmiI GGNNCC 2 cut(s) 32, 544
BmrFI CCNGG 5 cut(s) 62, 284, 386, 1178, 1336
BmsI GCATC 4 cut(s) 832, 1051, 1526, 1927
BpiI GAAGAC 5 cut(s) 108, 419, 723, 1370, 1455
BpmI CTGGAG 2 cut(s) 1076, 1829
Bpu10I CCTNAGC 1 cut(s) 1530
Bpu14I TTCGAA 1 cut(s) 686
BpuEI CTTGAG 2 cut(s) 1397, 1883
BpuMI CCSGG 2 cut(s) 62, 1178
BsaHI GRCGYC 1 cut(s) 105
BsaJI CCNNGG 2 cut(s) 6, 1671
BsaXI ACNNNNNCTCC 4 cut(s) 872, 902, 1872, 1902
Bsc4I CCNNNNNNNGG 3 cut(s) 27, 385, 1183
Bse1I ACTGG 1 cut(s) 779
BseBI CCWGG 3 cut(s) 284, 386, 1336
BseDI CCNNGG 2 cut(s) 6, 1671
BseGI GGATG 4 cut(s) 1042, 1432, 1517, 1725
BseLI CCNNNNNNNGG 3 cut(s) 27, 385, 1183
BseMII CTCAG 2 cut(s) 570, 687
BseNI ACTGG 1 cut(s) 779
BseRI GAGGAG 1 cut(s) 223
BseSI GKGCMC 2 cut(s) 529, 1737
BseXI GCAGC 8 cut(s) 58, 61, 64, 67, 184, 971, 1009, 1217
BsgI GTGCAG 2 cut(s) 970, 1316
Bsh1236I CGCG 2 cut(s) 739, 1026
BshFI GGCC 2 cut(s) 5, 11
BshNI GGYRCC 1 cut(s) 542
BsiHKAI GWGCWC 4 cut(s) 629, 702, 1092, 1737
BsiSI CCGG 2 cut(s) 62, 1177
BslFI GGGAC 1 cut(s) 472
BslI CCNNNNNNNGG 3 cut(s) 27, 385, 1183
BsmAI GTCTC 7 cut(s) 133, 356, 450, 575, 1127, 1445, 1590
BsmBI CGTCTC 2 cut(s) 133, 1445
BsmFI GGGAC 1 cut(s) 472
BsmI GAATGC 3 cut(s) 278, 1232, 1395
BsnI GGCC 2 cut(s) 5, 11
Bsp119I TTCGAA 1 cut(s) 686
Bsp1286I GDGCHC 7 cut(s) 75, 529, 629, 702, 750, 1092, 1737
Bsp143I GATC 9 cut(s) 30, 151, 160, 187, 423, 1146, 1217, 1650, 1784
BspACI CCGC 8 cut(s) 15, 44, 108, 122, 138, 878, 1024, 1779
BspANI GGCC 2 cut(s) 5, 11
BspCNI CTCAG 2 cut(s) 571, 688
BspFNI CGCG 2 cut(s) 739, 1026
BspHI TCATGA 1 cut(s) 1003
BspLI GGNNCC 2 cut(s) 32, 544
BspPI GGATC 5 cut(s) 25, 38, 155, 431, 1225
BspT104I TTCGAA 1 cut(s) 686
BspT107I GGYRCC 1 cut(s) 542
BsrBI CCGCTC 1 cut(s) 124
BsrI ACTGG 1 cut(s) 779
BssECI CCNNGG 2 cut(s) 6, 1671
BssMI GATC 9 cut(s) 30, 151, 160, 187, 423, 1146, 1217, 1650, 1784
BssNI GRCGYC 1 cut(s) 105
Bst2UI CCWGG 3 cut(s) 284, 386, 1336
Bst4CI ACNGT 5 cut(s) 962, 1118, 1276, 1405, 1456
Bst6I CTCTTC 2 cut(s) 210, 1383
BstACI GRCGYC 1 cut(s) 105
BstBI TTCGAA 1 cut(s) 686
BstC8I GCNNGC 1 cut(s) 1871
BstDEI CTNAG 4 cut(s) 579, 637, 696, 1530
BstDSI CCRYGG 2 cut(s) 6, 1671
BstF5I GGATG 4 cut(s) 1042, 1432, 1517, 1725
BstFNI CGCG 2 cut(s) 739, 1026
BstKTI GATC 9 cut(s) 33, 154, 163, 190, 426, 1149, 1220, 1653, 1787
BstMAI GTCTC 7 cut(s) 133, 356, 450, 575, 1127, 1445, 1590
BstMBI GATC 9 cut(s) 30, 151, 160, 187, 423, 1146, 1217, 1650, 1784
BstMWI GCNNNNNNNGC 9 cut(s) 49, 52, 55, 251, 416, 633, 745, 1709, 1813
BstNI CCWGG 3 cut(s) 284, 386, 1336
BstNSI RCATGY 3 cut(s) 399, 1873, 1886
BstSCI CCNGG 5 cut(s) 60, 282, 384, 1176, 1334
BstSFI CTRYAG 2 cut(s) 555, 1437
BstSLI GKGCMC 2 cut(s) 529, 1737
BstUI CGCG 2 cut(s) 739, 1026
BstV1I GCAGC 8 cut(s) 58, 61, 64, 67, 184, 971, 1009, 1217
BstV2I GAAGAC 5 cut(s) 108, 419, 723, 1370, 1455
BstX2I RGATCY 3 cut(s) 30, 423, 1784
BstXI CCANNNNNNTGG 1 cut(s) 599
BstYI RGATCY 3 cut(s) 30, 423, 1784
BsuI GTATCC 1 cut(s) 1494
BsuRI GGCC 2 cut(s) 5, 11
BtgI CCRYGG 2 cut(s) 6, 1671
BtgZI GCGATG 2 cut(s) 720, 1041
BtsCI GGATG 4 cut(s) 1042, 1432, 1517, 1725
BtsI GCAGTG 1 cut(s) 1774
BtsIMutI CAGTG 2 cut(s) 576, 1774
Cac8I GCNNGC 1 cut(s) 1871
CciI TCATGA 1 cut(s) 1003
CseI GACGC 3 cut(s) 113, 465, 728
Csp6I GTAC 3 cut(s) 313, 466, 1106
CspCI CAANNNNNGTGG 2 cut(s) 1743, 1778
CviQI GTAC 3 cut(s) 313, 466, 1106
DdeI CTNAG 4 cut(s) 579, 637, 696, 1530
DpnI GATC 9 cut(s) 32, 153, 162, 189, 425, 1148, 1219, 1652, 1786
DpnII GATC 9 cut(s) 30, 151, 160, 187, 423, 1146, 1217, 1650, 1784
DrdI GACNNNNNNGTC 1 cut(s) 1459
DriI GACNNNNNGTC 1 cut(s) 738
DseDI GACNNNNNNGTC 1 cut(s) 1459
EaeI YGGCCR 1 cut(s) 3
Eam1104I CTCTTC 2 cut(s) 210, 1383
Eam1105I GACNNNNNGTC 1 cut(s) 738
EarI CTCTTC 2 cut(s) 210, 1383
EciI GGCGGA 2 cut(s) 4, 127
Ecl136II GAGCTC 3 cut(s) 627, 700, 1090
Eco24I GRGCYC 5 cut(s) 75, 629, 702, 750, 1092
Eco32I GATATC 1 cut(s) 1512
Eco53kI GAGCTC 3 cut(s) 627, 700, 1090
Eco57I CTGAAG 2 cut(s) 267, 1812
EcoICRI GAGCTC 3 cut(s) 627, 700, 1090
EcoRII CCWGG 3 cut(s) 282, 384, 1334
EcoRV GATATC 1 cut(s) 1512
EcoT38I GRGCYC 5 cut(s) 75, 629, 702, 750, 1092
Esp3I CGTCTC 2 cut(s) 133, 1445
FaqI GGGAC 1 cut(s) 472
FauI CCCGC 2 cut(s) 1031, 1772
FauNDI CATATG 1 cut(s) 309
FbaI TGATCA 1 cut(s) 1650
FblI GTMKAC 1 cut(s) 1301
FokI GGATG 4 cut(s) 1029, 1419, 1504, 1712
FriOI GRGCYC 5 cut(s) 75, 629, 702, 750, 1092
FspBI CTAG 5 cut(s) 84, 1485, 1497, 1709, 1713
GsuI CTGGAG 2 cut(s) 1076, 1829
HaeIII GGCC 2 cut(s) 5, 11
HapII CCGG 2 cut(s) 62, 1177
HgaI GACGC 3 cut(s) 113, 465, 728
Hin1I GRCGYC 1 cut(s) 105
HincII GTYRAC 1 cut(s) 732
HindII GTYRAC 1 cut(s) 732
HindIII AAGCTT 1 cut(s) 498
HpaII CCGG 2 cut(s) 62, 1177
HphI GGTGA 2 cut(s) 111, 803
Hpy166II GTNNAC 7 cut(s) 732, 794, 1108, 1302, 1735, 1747, 1822
Hpy8I GTNNAC 7 cut(s) 732, 794, 1108, 1302, 1735, 1747, 1822
Hpy99I CGWCG 1 cut(s) 107
HpyAV CCTTC 3 cut(s) 174, 399, 807
HpyCH4III ACNGT 5 cut(s) 962, 1118, 1276, 1405, 1456
HpyCH4IV ACGT 2 cut(s) 815, 1304
HpyF10VI GCNNNNNNNGC 9 cut(s) 49, 52, 55, 251, 416, 633, 745, 1709, 1813
HpyF3I CTNAG 4 cut(s) 579, 637, 696, 1530
HpySE526I ACGT 2 cut(s) 815, 1304
Hsp92I GRCGYC 1 cut(s) 105
Ksp22I TGATCA 1 cut(s) 1650
Kzo9I GATC 9 cut(s) 30, 151, 160, 187, 423, 1146, 1217, 1650, 1784
LmnI GCTCC 5 cut(s) 140, 161, 705, 1087, 1095
Lsp1109I GCAGC 8 cut(s) 58, 61, 64, 67, 184, 971, 1009, 1217
LweI GCATC 4 cut(s) 832, 1051, 1526, 1927
MaeI CTAG 5 cut(s) 84, 1485, 1497, 1709, 1713
MaeII ACGT 2 cut(s) 815, 1304
MaeIII GTNAC 3 cut(s) 37, 391, 550
MalI GATC 9 cut(s) 32, 153, 162, 189, 425, 1148, 1219, 1652, 1786
MbiI CCGCTC 1 cut(s) 124
MboI GATC 9 cut(s) 30, 151, 160, 187, 423, 1146, 1217, 1650, 1784
MfeI CAATTG 1 cut(s) 271
MflI RGATCY 3 cut(s) 30, 423, 1784
MhlI GDGCHC 7 cut(s) 75, 529, 629, 702, 750, 1092, 1737
MlyI GAGTC 2 cut(s) 92, 890
MmeI TCCRAC 3 cut(s) 126, 433, 1287
MroXI GAANNNNTTC 2 cut(s) 467, 780
MseI TTAA 4 cut(s) 642, 1050, 1830, 1926
MslI CAYNNNNRTG 3 cut(s) 1728, 1730, 1881
MspA1I CMGCKG 2 cut(s) 245, 984
MspI CCGG 2 cut(s) 62, 1177
MspR9I CCNGG 5 cut(s) 62, 284, 386, 1178, 1336
MunI CAATTG 1 cut(s) 271
Mva1269I GAATGC 3 cut(s) 278, 1232, 1395
MvaI CCWGG 3 cut(s) 284, 386, 1336
MvnI CGCG 2 cut(s) 739, 1026
MwoI GCNNNNNNNGC 9 cut(s) 49, 52, 55, 251, 416, 633, 745, 1709, 1813
NciI CCSGG 2 cut(s) 62, 1178
NdeI CATATG 1 cut(s) 309
NdeII GATC 9 cut(s) 30, 151, 160, 187, 423, 1146, 1217, 1650, 1784
NlaIV GGNNCC 2 cut(s) 32, 544
NmeAIII GCCGAG 1 cut(s) 1837
NmuCI GTSAC 2 cut(s) 37, 391
NspI RCATGY 3 cut(s) 399, 1873, 1886
NspV TTCGAA 1 cut(s) 686
PaeI GCATGC 1 cut(s) 1873
PagI TCATGA 1 cut(s) 1003
PcsI WCGNNNNNNNCGW 1 cut(s) 890
PctI GAATGC 3 cut(s) 278, 1232, 1395
PdmI GAANNNNTTC 2 cut(s) 467, 780
PfeI GAWTC 9 cut(s) 87, 507, 569, 653, 692, 722, 836, 947, 1630
PfoI TCCNGGA 2 cut(s) 282, 384
PleI GAGTC 2 cut(s) 92, 889
PpsI GAGTC 2 cut(s) 92, 889
Psp124BI GAGCTC 3 cut(s) 629, 702, 1092
Psp6I CCWGG 3 cut(s) 282, 384, 1334
PspGI CCWGG 3 cut(s) 282, 384, 1334
PspN4I GGNNCC 2 cut(s) 32, 544
PsuI RGATCY 3 cut(s) 30, 423, 1784
PvuII CAGCTG 2 cut(s) 245, 984
RsaI GTAC 3 cut(s) 314, 467, 1107
RsaNI GTAC 3 cut(s) 313, 466, 1106
RseI CAYNNNNRTG 3 cut(s) 1728, 1730, 1881
SacI GAGCTC 3 cut(s) 629, 702, 1092
SaqAI TTAA 4 cut(s) 642, 1050, 1830, 1926
Sau3AI GATC 9 cut(s) 30, 151, 160, 187, 423, 1146, 1217, 1650, 1784
ScaI AGTACT 1 cut(s) 467
SchI GAGTC 2 cut(s) 92, 890
ScrFI CCNGG 5 cut(s) 62, 284, 386, 1178, 1336
SduI GDGCHC 7 cut(s) 75, 529, 629, 702, 750, 1092, 1737
SfaNI GCATC 4 cut(s) 832, 1051, 1526, 1927
SfcI CTRYAG 2 cut(s) 555, 1437
SfuI TTCGAA 1 cut(s) 686
SmiMI CAYNNNNRTG 3 cut(s) 1728, 1730, 1881
SmlI CTYRAG 2 cut(s) 1376, 1862
SmoI CTYRAG 2 cut(s) 1376, 1862
SphI GCATGC 1 cut(s) 1873
SsiI CCGC 8 cut(s) 15, 44, 108, 122, 138, 878, 1024, 1779
SspMI CTAG 5 cut(s) 84, 1485, 1497, 1709, 1713
SstI GAGCTC 3 cut(s) 629, 702, 1092
StyD4I CCNGG 5 cut(s) 60, 282, 384, 1176, 1334
TaaI ACNGT 5 cut(s) 962, 1118, 1276, 1405, 1456
TaiI ACGT 2 cut(s) 818, 1307
TaqI TCGA 7 cut(s) 69, 261, 629, 686, 1102, 1131, 1318
TatI WGTACW 2 cut(s) 465, 1105
TauI GCSGC 2 cut(s) 46, 110
TfiI GAWTC 9 cut(s) 87, 507, 569, 653, 692, 722, 836, 947, 1630
Tru1I TTAA 4 cut(s) 642, 1050, 1830, 1926
Tru9I TTAA 4 cut(s) 642, 1050, 1830, 1926
TscAI CASTG 2 cut(s) 583, 1774
TseFI GTSAC 2 cut(s) 37, 391
TseI GCWGC 8 cut(s) 46, 49, 52, 55, 197, 984, 997, 1230
Tsp45I GTSAC 2 cut(s) 37, 391
TspRI CASTG 2 cut(s) 583, 1774
VneI GTGCAC 1 cut(s) 1733
XapI RAATTY 6 cut(s) 444, 681, 860, 1070, 1097, 1610
XbaI TCTAGA 1 cut(s) 83
XceI RCATGY 3 cut(s) 399, 1873, 1886
XcmI CCANNNNNNNNNTGG 1 cut(s) 962
XmiI GTMKAC 1 cut(s) 1301
XmnI GAANNNNTTC 2 cut(s) 467, 780
XspI CTAG 5 cut(s) 84, 1485, 1497, 1709, 1713
ZrmI AGTACT 1 cut(s) 467
Using CommOnly database (standard laboratory enzymes). Scanned on CDS sequence.