Restriction Enzymes that DO NOT CUT in this sequence AatII AccI AceIII AflII AflIII AhdI AluI Alw26I ApaI ApaBI ApaLI ApoI AscI AvaI AvaII AvrII BaeI BamHI BanI BanII BbsI Bce83I BcefI BcgI BcgI BclI BfaI BfiI BglI BglII BmgI BplI BpmI Bpu10I Bpu1102I BsaI BsaAI BsaBI BsaHI BsaJI BsaWI BsaXI BscGI BseRI BsgI BsiHKAI BslI BsmBI BsmFI Bsp24I Bsp24I Bsp1286I BspEI BspGI BspLU11I BsrBI BsrDI BsrFI BsrGI BssHII BssSI Bst1107I BstEII BstXI BstYI Bsu36I Cac8I CjeI CjeI CjePI CjePI ClaI DdeI DraI DraIII DrdI DrdII DsaI EaeI EagI EarI EciI Eco47III Eco57I EcoNI EcoO109I EcoRI EcoRII EcoRV FokI FseI FspI GdiII HaeI HaeIII HgaI HgiEII Hin4I HincII HindIII HpaI KpnI MaeIII MluI MmeI MnlI MscI MslI MunI MwoI NarI NcoI NdeI NgoAIV NheI NlaIV NotI NruI NsiI NspI NspV PacI PflMI PinAI PleI PmeI PmlI PpuMI PshAI Psp1406I PstI PvuI PvuII RcaI RleAI RsaI RsrII SacII SalI SanDI SapI Sau96I ScaI SexAI SfcI SfiI SgfI SgrAI SimI SmaI SnaBI SpeI SphI SrfI Sse8387I Sse8647I SspI SstI StuI StyI SunI SwaI TaqII TaqII TatI ThaI Tsp45I Tsp509I TspRI Tth111I Tth111II UbaDI VspI XbaI XcmI XhoI XmnI Total Number of Cuts per Restriction Enzyme AatII 0 BsaHI 0 DraI 0 MslI 0 SexAI 0 AccI 0 BsaJI 0 DraIII 0 MspI 2 SfaNI 1 AceIII 0 BsaWI 0 DrdI 0 MspA1I 1 SfcI 0 AciI 2 BsaXI 0 DrdII 0 MunI 0 SfiI 0 AflII 0 BsbI 1 DsaI 0 MwoI 0 SgfI 0 AflIII 0 BscGI 0 EaeI 0 NarI 0 SgrAI 0 AhdI 0 BseRI 0 EagI 0 NciI 2 SimI 0 AluI 0 BsgI 0 EarI 0 NcoI 0 SmaI 0 AlwI 1 BsiEI 1 EciI 0 NdeI 0 SnaBI 0 Alw26I 0 BsiHKAI 0 Eco47III 0 NgoAIV 0 SpeI 0 AlwNI 1 BslI 0 Eco57I 0 NheI 0 SphI 0 ApaI 0 BsmI 1 EcoNI 0 NlaIII 1 SrfI 0 ApaBI 0 BsmBI 0 EcoO109I 0 NlaIV 0 Sse8387I 0 ApaLI 0 BsmFI 0 EcoRI 0 NotI 0 Sse8647I 0 ApoI 0 Bsp24I 0 EcoRII 0 NruI 0 SspI 0 AscI 0 Bsp24I 0 EcoRV 0 NsiI 0 SstI 0 AvaI 0 Bsp1286I 0 FauI 1 NspI 0 StuI 0 AvaII 0 BspEI 0 Fnu4HI 2 NspV 0 StyI 0 AvrII 0 BspGI 0 FokI 0 PacI 0 SunI 0 BaeI 0 BspLU11I 0 FseI 0 Pfl1108I 2 SwaI 0 BamHI 0 BspMI 1 FspI 0 PflMI 0 TaqI 2 BanI 0 BsrI 1 GdiII 0 PinAI 0 TaqII 0 BanII 0 BsrBI 0 HaeI 0 PleI 0 TaqII 0 BbsI 0 BsrDI 0 HaeII 1 PmeI 0 TatI 0 BbvI 1 BsrFI 0 HaeIII 0 PmlI 0 TauI 1 BccI 1 BsrGI 0 HgaI 0 PpuMI 0 TfiI 1 Bce83I 0 BssHII 0 HgiEII 0 PshAI 0 ThaI 0 BcefI 0 BssSI 0 HhaI 1 Psp1406I 0 TseI 1 BcgI 0 Bst1107I 0 Hin4I 0 PstI 0 Tsp45I 0 BcgI 0 BstEII 0 HincII 0 PvuI 0 Tsp4CI 1 BclI 0 BstXI 0 HindIII 0 PvuII 0 Tsp509I 0 BfaI 0 BstYI 0 HinfI 1 RcaI 0 TspRI 0 BfiI 0 Bsu36I 0 HpaI 0 RleAI 0 Tth111I 0 BglI 0 Cac8I 0 HphI 1 RsaI 0 Tth111II 0 BglII 0 CjeI 0 KpnI 0 RsrII 0 UbaDI 0 BmgI 0 CjeI 0 MaeII 4 SacII 0 VspI 0 BplI 0 CjePI 0 MaeIII 0 SalI 0 XbaI 0 BpmI 0 CjePI 0 MboII 1 SanDI 0 XcmI 0 Bpu10I 0 ClaI 0 MluI 0 SapI 0 XhoI 0 Bpu1102I 0 CviJI 2 MmeI 0 Sau96I 0 XmnI 0 BsaI 0 CviRI 3 MnlI 0 Sau3AI 3 BsaAI 0 DdeI 0 MscI 0 ScaI 0 BsaBI 0 DpnI 3 MseI 1 ScrFI 2 Cut Sites by Restriction Enzyme AciI C'CG_C - 2 Cut(s) 44 234 AlwI GGATCnnnn'n_ - 1 Cut(s) 35 AlwNI CAG_nnn'CTG - 1 Cut(s) 213 BbvI GCAGCnnnnnnnn'nnnn_ - 1 Cut(s) 171 BccI CCATC - 1 Cut(s) 7 BsbI CAACAC - 1 Cut(s) 186 BsiEI CG_ry'CG - 1 Cut(s) 193 BsmI GAATG_Cn' - 1 Cut(s) 32 BspMI ACCTGCnnnn'nnnn_ - 1 Cut(s) 68 BsrI ACTG_Gn' - 1 Cut(s) 197 CviRI TG'CA - 3 Cut(s) 30 135 183 DpnI GA'TC - 3 Cut(s) 41 164 217 FauI CCCGCnnnn'nn_ - 1 Cut(s) 51 Fnu4HI GC'n_GC - 2 Cut(s) 181 234 HaeII r_GCGC'y - 1 Cut(s) 213 HhaI G_CG'C - 1 Cut(s) 212 HinfI G'AnT_C - 1 Cut(s) 152 HphI GGTGAnnnnnnn_n' - 1 Cut(s) 83 MaeII A'CG_T - 4 Cut(s) 15 127 141 174 MboII GAAGAnnnnnnn_n' - 1 Cut(s) 146 MseI T'TA_A - 1 Cut(s) 168 MspI C'CG_G - 2 Cut(s) 65 149 MspA1I CmG'CkG - 1 Cut(s) 46 NciI CC's_GG - 2 Cut(s) 65 149 NlaIII _CATG' - 1 Cut(s) 100 Pfl1108I TCGTAG - 2 Cut(s) 54 87 Sau3AI 'GATC_ - 3 Cut(s) 39 162 215 ScrFI CC'n_GG - 2 Cut(s) 65 149 SfaNI GCATCnnnnn'nnnn_ - 1 Cut(s) 144 TaqI T'CG_A - 2 Cut(s) 138 222 TauI GCsGC - 1 Cut(s) 234 TfiI G'AwT_C - 1 Cut(s) 152 TseI G'CwG_C - 1 Cut(s) 180 Tsp4CI AC_n'GT - 1 Cut(s) 194 Restriction Map of Sequence AlwI AciI CviRI Sau3AI MspA1I Pfl1108I BccI MaeII BsmI DpnI FauI \ \ \ \ \ \ \ \ \ \ \ 1 aaaaccatcaaatacgttatgctggaatgcaacgaactgatcccgctgttctacgaaacc 60 ttttggtagtttatgcaatacgaccttacgttgcttgactagggcgacaagatgctttgg ^ * ^ * ^ * ^ * ^ * ^ * K T I K Y V M L E C N E L I P L F Y E T CviJI MspI ScrFI NciI Pfl1108I BspMI HphI NlaIII \ \ \ \ \ 61 tgcccggctggtgaaaacatctgctacgaaatgttcatggttgctaccccgaaagttccg 120 acgggccgaccacttttgtagacgatgctttacaagtaccaacgatggggctttcaaggc ^ * ^ * ^ * ^ * ^ * ^ * C P A G E N I C Y E M F M V A T P K V P MboII TfiI MspI MaeII ScrFI MseI TaqI NciI Sau3AI BbvI TseI MaeII CviRI SfaNI HinfI DpnI MaeII Fnu4HI \ \ \ \ \ \ \ \ \ \ \ \ \ \\ 121 tgcgaacgtggttgcatcgacgtttgcccggaatcttctctgatcgttaaatacgtttgc 180 acgcttgcaccaacgtagctgcaaacgggccttagaagagactagcaatttatgcaaacg ^ * ^ * ^ * ^ * ^ * ^ * C E R G C I D V C P E S S L I V K Y V C Sau3AI AciI BsrI HhaI DpnI TauI BsbI Tsp4CI HaeII Fnu4HI CviRI BsiEI AlwNI TaqI CviJI \ \ \\ \ \\ \ \ \ \\ 181 tgcaacaccgaccgttgccagtaatccagcgcctgatctctcgaaataaaagccgcatt 240 acgttgtggctggcaacggtcattaggtcgcggactagagagctttattttcggcgtaa ^ * ^ * ^ * ^ * ^ * ^ * C N T D R C Q Z S S A Z S L E I K A A Ladder Map of Restriction Enzyme Cut Sites 20 40 60 80 100 120 140 160 180 200 220 240 : : : : : : : : : : : : AciI -------------------------\-----------------------------------------------------------------------------------------------------------------\---- AlwI --------------------\--------------------------------------------------------------------------------------------------------------------------- AlwNI -------------------------------------------------------------------------------------------------------------------------------/---------------- BbvI ------------------------------------------------------------------------------------------------------\----------------------------------------- : : : : : : : : : : : : BccI ---|-------------------------------------------------------------------------------------------------------------------------------------------- BsbI ---------------------------------------------------------------------------------------------------------------|-------------------------------- BsiEI -------------------------------------------------------------------------------------------------------------------/---------------------------- BsmI ------------------/----------------------------------------------------------------------------------------------------------------------------- : : : : : : : : : : : : BspMI ----------------------------------------\------------------------------------------------------------------------------------------------------- BsrI ---------------------------------------------------------------------------------------------------------------------/-------------------------- CviRI -----------------|--------------------------------------------------------------|----------------------------|---------------------------------- DpnI ------------------------|------------------------------------------------------------------------|-------------------------------|-------------- : : : : : : : : : : : : FauI ------------------------------\----------------------------------------------------------------------------------------------------------------- Fnu4HI ------------------------------------------------------------------------------------------------------------\------------------------------\---- HaeII -------------------------------------------------------------------------------------------------------------------------------/---------------- HhaI ------------------------------------------------------------------------------------------------------------------------------/----------------- : : : : : : : : : : : : HinfI ------------------------------------------------------------------------------------------\----------------------------------------------------- HphI -------------------------------------------------/---------------------------------------------------------------------------------------------- MaeII --------\------------------------------------------------------------------\--------\------------------\---------------------------------------- MboII ---------------------------------------------------------------------------------------/-------------------------------------------------------- 20 40 60 80 100 120 140 160 180 200 220 240 : : : : : : : : : : : : MseI ----------------------------------------------------------------------------------------------------\------------------------------------------- MspI --------------------------------------\-------------------------------------------------\------------------------------------------------------- MspA1I ---------------------------|-------------------------------------------------------------------------------------------------------------------- NciI --------------------------------------\-------------------------------------------------\------------------------------------------------------- : : : : : : : : : : : : NlaIII -----------------------------------------------------------/------------------------------------------------------------------------------------ Pfl1108I -------------------------------|-------------------|-------------------------------------------------------------------------------------------- Sau3AI ----------------------\-------------------------------------------------------------------------\-------------------------------\--------------- ScrFI --------------------------------------\-------------------------------------------------\------------------------------------------------------- : : : : : : : : : : : : SfaNI -------------------------------------------------------------------------------------\---------------------------------------------------------- TaqI ----------------------------------------------------------------------------------\-------------------------------------------------\----------- TauI -------------------------------------------------------------------------------------------------------------------------------------------|---- TfiI ------------------------------------------------------------------------------------------\----------------------------------------------------- : : : : : : : : : : : : TseI -----------------------------------------------------------------------------------------------------------\------------------------------------ Tsp4CI -------------------------------------------------------------------------------------------------------------------/---------------------------- Summary of Enzymes that cut ** 2 ** times or less TfiI@152 TaqI@138 ScrFI@65 SfaNI@144 Pfl1108I@54 ScrFI@149 Tsp4CI@194 TauI@234 NciI@65 NciI@149 Fnu4HI@181 TaqI@222 BsmI@32 MspA1I@46 MspI@65 MspI@149 TseI@180 BsrI@197 HhaI@212 Fnu4HI@234 AlwI@35 FauI@51 CviJI@68 Pfl1108I@87 MboII@146 MseI@168 BsiEI@193 HaeII@213 CviJI@233 BccI@7 AciI@44 BspMI@68 HphI@83 NlaIII@100 HinfI@152 BbvI@171 BsbI@186 AlwNI@213 AciI@234 | | | | | || | | | | | | | || | | | || | | | || | | ------------------------------------------------------------------------------------------------------------------------------------------------ : : : : : : : : : : : : 20 40 60 80 100 120 140 160 180 200 220 240 Pseudo-Gel Map of Digestions 10 100 . . . . . . . . . . . AciI | | | AlwI | | AlwNI | | BbvI | | . . . . . . . . . . . BccI | | BsbI | | BsiEI | | BsmI | | . . . . . . . . . . . BspMI | | BsrI | | CviRI | | | | DpnI | | | | . . . . . . . . . . . FauI | | Fnu4HI | | | HaeII | | HhaI | | . . . . . . . . . . . HinfI | | HphI | | MaeII | | | | | MboII | | 10 100 . . . . . . . . . . . MseI | | MspI | | | MspA1I | | NciI | | | . . . . . . . . . . . NlaIII | | Pfl1108I | | | Sau3AI | | | | ScrFI | | | . . . . . . . . . . . SfaNI | | TaqI | | | TauI | | TfiI | | . . . . . . . . . . . TseI | | Tsp4CI | |