5’ - TCAGATAAAAAACTGGCACGCAATCTGCAATTAGCAAGACATCTTTTTAGAACACGCTGAATAAATTGAGGTTGC

Question

5’ - TCAGATAAAAAACTGGCACGCAATCTGCAATTAGCAAGACATCTTTTTAGAACACGCTGAATAAATTGAGGTTGCTATGTCTATTGTGGTGAAAAATAACATTCATTGGGTTGGTCAACGTGACTGGGAAGTGCGTGATTTTCACGGCACGGAATATAAAACGCTGCGCGGCAGCAGCTACAATAGCTACCTCATCCGCGAAGAAAAAAACGTGCTGATCGACACCGTCGACCATAAATTCAGCCGCGAATTTGTGCAGAACCTGCGTAATGAAATCGATCTGGCGGATATCGATTACATCGTGATTAACCATGCAGAAGAGGACCACGCTGGGGCGCTGACCGAACTGATGGCACAAATTCCCGATACGCCGATCTACTGTACAGCCAACGCTATCGACTCGATAAATGGTCATCACCATCATCCGGAGTGGAATTTTAATGTGGTGAAAACTGGCGACACGCTGGATATCGGCAACGGCAAACAGCTCATTTTTGTCGAAACACCAATGCTGCACTGGCCGGACAGCATGATGACTTACCTGACAGGCGACGCGGTGCTGTTCAGTAACGATGCTTTCGGTCAACACTACTGCGACGAGCATCTGTTCAACGATGAAGTGGATCAGACGGAGCTTTTCGAGCAGTGCCAGCGTTACTACGCCAATATCCTGACGCCGTTCAGCCGCCTGGTAACACCGAAAATTACCGAGATCCTGGGCTTTAACTTACCAGTCGATATGATAGCCACTTCCCACGGCGTGGTATGGCGCGATAACCCGACGCAAATTGTCGAGCTGTACCTGAAATGGGCGGCTGATTATCAGGAAGACAGAATCACCATTTTCTACGACACCATGTCGAATAACACCCGCATGATGGCTGACGCTATCGCCCAGGGGATTGCGGAAACCGACCCACGCGTGGCGGTGAAAATTTTCAACGTCGCCCGAAGCGATAAAAACGAAATCCTGACTAATGTCTTCCGCTCAAAAGGCGTGCTGGTCGGCACTTCGACGATGAATAACGTGATGATGCCGAAAATCGCCGGGCTGGTGGAGGAGATGACTGGTTTACGCTTCCGTAACAAACGCGCCAGTGCTTTCGGCTCTCACGGCTGGAGCGGCGGTGCGGTGGATCGTCTTTCCACGCGCCTGCAGGATGCGGGTTTCGAAATGTCGCTTAGCCTGAAAGCGAAATGGCGACCAGACCAGGACGCTCTGAAGTTATGCCGTGAACACGGTCGCGAAATCGCCCGTCAGTGGGCGCTCGCGCCGCTGCCGCAGAGCACGGTGAATACGGTAGTTAAAGAAGAAACCTCTGCCACCACGACGGCTGACCTCGGCCCACGGATGCAGTGCAGCGTCTGCCAGTGGATTTACGATCCGGCAAAAGGCGAGCCAATGCAGGACGTTGCGCCAGGAACGCCGTGGAGTGAAGTCCCGGATAACTTCCTCTGCCCGGAATGCTCCCTCGGCAAAGACGTCTTTGAAGAACTGGCATCGGAGGCAAAATGAGTAACGGCATTGTGATCATCGGTTCGGGCTTCGCCGCCCGCCAACTGGTGAAAAATATTCGCAAACAGGACGCCACTATTCCATT-3

4. Use an online tool to identify the sites within the coding region of the DNA sequence whichwould be subjected to restriction enzyme digestion. Please attach the detailed list ofrestriction enzyme sites located within this region of the DNA

Explanation / Answer

Following results were obtained when used online tool "RESTRICTION MAPPER" for the given sequence

Conformation: linear

Overhang: five_prime, three_prime, blunt

Minimum Site Length: 5 bases

Maximum Number of Cuts: all

Included: all commercial, prototypes only

Noncutters: AarI, AbsI, AclI, AflII, AgeI, AjuI, AloI, ApaI, ApaLI, ArsI, AscI, AvaI, AvrII, BaeI, BalI, BamHI, BarI, BbvCI, BcgI, BciVI, BglI, BglII, BplI, Bpu10I, BsaAI, BsaBI, BseMII, BsePI, BseSI, BsmAI, BspHI, BsrDI, BstEII, BstXI, BtgZI, BtrI, Cfr10I, CspCI, DraII, DrdI, Eco31I, Eco47III, EcoNI, EcoRI, Esp3I, FalI, FseI, FspAI, HindIII, HpaI, KpnI, MauBI, MfeI, MmeI, NaeI, NarI, NcoI, NdeI, NheI, NotI, NspI, OliI, PacI, PmaCI, PmeI, PpuMI, PsiI, PI-PspI, PspXI, PsrI, PvuI, PvuII, RsrII, SacI, SacII, SanDI, SapI, ScaI, PI-SceI, SexAI, SfiI, SgfI, SgrAI, SgrDI, SmaI, SmlI, SnaBI, SpeI, SphI, SrfI, StuI, StyI, SwaI, VspI, XbaI, XhoI

Name Sequence Site Length Overhang Frequency Cut Positions AatII GACGTC 6 three_prime 1 1485 AccI GTMKAC 6 five_prime 1 228 AcyI GRCGYC 6 five_prime 3 674, 1482, 1586 AflIII ACRYGT 6 five_prime 1 919 AgsI TTSAA 5 three_prime 3 610, 940, 1490 AlfI GCANNNNNNTGC 6 three_prime 2 1394, 1428 AlwNI CAGNNNCTG 6 three_prime 2 262, 1365 ApoI RAATTY 6 five_prime 5 236, 248, 357, 433, 933 AsuII TTCGAA 6 five_prime 1 1170 AvaII GGWCC 5 five_prime 1 322 BbvI GCAGC 5 five_prime 6 150, 183, 186, 498, 1263, 1371 BccI CCATC 5 five_prime 3 343, 426, 871 BclI TGATCA 6 five_prime 1 1529 BdaI TGANNNNNNTCA 6 three_prime 6 605, 639, 661, 695, 919, 953 BfiI ACTGGG 6 three_prime 1 133 BsaXI ACNNNNNCTCC 6 three_prime 2 1111, 1141 BseRI GAGGAG 6 three_prime 1 1073 BseYI CCCAGC 6 five_prime 1 329 BsgI GTGCAG 6 three_prime 3 275, 497, 1378 BsmI GAATGC 6 three_prime 1 1469 Bsp1407I TGTACA 6 five_prime 1 380 BspMI ACCTGC 6 five_prime 1 270 BsrBI CCGCTC 6 blunt 2 987, 1122 BsrI ACTGG 5 three_prime 10 17, 128, 457, 521, 731, 1072, 1095, 1369, 1500, 1565 BtsI GCAGTG 6 three_prime 2 650, 1361 CfrI YGGCCR 6 five_prime 1 518 ClaI ATCGAT 6 five_prime 2 276, 291 DraIII CACNNNGTG 6 three_prime 1 760 Eam1105I GACNNNNNGTC 6 three_prime 1 856 EciI GGCGGA 6 three_prime 1 299 Eco57I CTGAAG 6 three_prime 1 1241 Eco57MI CTGRAG 6 three_prime 2 1138, 1241 EcoP15I CAGCAG 6 five_prime 1 202 EcoRII CCWGG 5 five_prime 5 687, 714, 894, 1209, 1417 EcoRV GATATC 6 blunt 2 289, 469 FauI CCCGC 5 five_prime 3 878, 1156, 1561 FokI GGATG 5 five_prime 4 179, 408, 1172, 1363 GsuI CTGGAG 6 three_prime 1 1138 HaeII RGCGCY 6 three_prime 2 338, 1268 HaeIV GAYNNNNNRTC 6 three_prime 4 393, 427, 875, 1399 HgaI GACGC 5 five_prime 7 560, 682, 790, 893, 1223, 1351, 1594 Hin4I GAYNNNNNVTC 6 three_prime 6 394, 426, 842, 874, 1366, 1398 HindII GTYRAC 6 blunt 3 116, 229, 584 HphI GGTGA 5 three_prime 7 100, 407, 457, 829, 940, 1303, 1575 Hpy99I CGWCG 5 three_prime 7 230, 554, 599, 784, 947, 1018, 1333 MboII GAAGA 5 three_prime 6 212, 329, 839, 973, 1322, 1502 MluI ACGCGT 6 five_prime 1 919 MslI CAYNNNNRTG 6 blunt 1 872 NmeAIII GCCGAG 6 three_prime 2 1320, 1452 NruI TCGCGA 6 blunt 1 1245 PasI CCCWGGG 7 five_prime 1 895 PflMI CCANNNNNTGG 6 three_prime 1 760 PfoI TCCNGGA 6 five_prime 1 1440 PleI GAGTC 5 five_prime 1 392 PpiI GAACNNNNNCTC 7 three_prime 2 590, 622 PshAI GACNNNNGTC 6 blunt 1 977 PstI CTGCAG 6 three_prime 1 1158 SalI GTCGAC 6 five_prime 1 227 SduI GDGCHC 6 three_prime 1 1289 SfaNI GCATC 5 five_prime 6 562, 610, 1023, 1150, 1341, 1508 Sse8387I CCTGCAGG 8 three_prime 1 1158 SspI AATATT 6 blunt 1 1572 TaqII GACCGA 6 three_prime 4 330, 356, 569, 595 TatI WGTACW 6 five_prime 1 380 TauI GCSGC 5 three_prime 8 171, 246, 687, 815, 1125, 1276, 1282, 1552 TfiI GAWTC 5 five_prime 1 834 TseI GCWGC 5 five_prime 6 163, 171, 174, 511, 1276, 1359 TsoI TARCCA 6 three_prime 2 323, 760 Tsp45I GTSAC 5 five_prime 1 119 TspDTI ATGAA 5 three_prime 4 92, 285, 630, 1034 TspGWI ACGGA 5 three_prime 4 164, 644, 1070, 1363 TspRI CASTG 5 three_prime 6 521, 650, 1102, 1265, 1361, 1376 TstI CACNNNNNNTCC 6 three_prime 2 1111, 1143 Tth111I GACNNNGTC 6 five_prime 1 224 XcmI CCANNNNNNNNNTGG 6 three_prime 2 513, 1425 XhoII RGATCY 6 five_prime 1 711 XmnI GAANNNNTTC 6 blunt 1 935

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