Mus musculus (mm10), chr6:52313498-52314271, forward genomic strand
Your input sequence is 774 bp long. It contains 161 possible guide sequences.
Shown below are their PAM sites and the expected cleavage position located -3bp 5' of the PAM site.
Click on a match for the PAM NGG below to show its 20 bp-long guide sequence. (Need help? Look at the CRISPOR manual)
Colors green, yellow and red indicate high, medium and low specificity of the PAM's guide sequence in the genome.

Variant database: Missing a variant database? We can add it.
Position  0         10        20        30        40        50        60        70        80        90        100       110       120       130       140       150       160       170       180       190       200       210       220       230       240       250       260       270       280       290       300       310       320       330       340       350       360       370       380       390       400       410       420       430       440       450       460       470       480       490       500       510       520       530       540       550       560       570       580       590       600       610       620       630       640       650       660       670       680       690       700       710       720       730       740       750       760       770       
Variants  ......................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................
Sequence  ctcctttcatcttcactgtggcagacgtttctatttatccacttgcgttcgccgagtggcgtcaccagcggtactgtaatgacgattgcagcaggaggatgacagcttagaaagaagagggcaatggggcttcctcccagaggcggtgcggcacagaggagcgctcgcatcacaaggtgaccctagctccccactgccatctccgcggtcgccgtcgacacggcgctggggctacccggcgcctgccttgtcgccttagctcctcttctcagccaagatcccagggagcctgggattaggagcttacttgggggttttccccctccccccctctggagagtccggggATGgagagccgaaaggacatggttatgtttctggatgggggtcagcttggcactctggttggtaagagggtctctaatttgtccgaagccgtgagcagcccgctgcctgaaccgccagagaagatggtgccccacggttgcctgagcccgcgagccggccctccgacttcccgggagcgtggcgggggaggccaggaggaggagccggtcgatggactagcaggcagtgctgcagggctgggcgccgagccacggtctgctggagcggccatgcttggcccgggacccccagtcccctccgcggacagcctctctggccaagggcaacctagtagctcagacaccgaatcggatttctatgaagaaatcgaggtgagctgcaccccagactgcgccaccgggaacgccgagtaccagcacagcaaag
            CCT---                              CCA---       CCG---       CCA---                   −−−AGG                   −−−AGG  −−−GGG    CCT---  −−−CGG       −−−AGG            −−−AGG   CCC---  CCC---  CCA---         CCG---      −−−TGG     CCC---     CCT---  CCT---  CCT---     CCA---  CCA---  CCT---           −−−TGG       CCC---  CCC---         CCG---        CCG---  −−−TGG      −−−TGG          −−−TGG  −−−TGG     −−−AGG             CCG---          CCC---       CCG---    −−−TGG  CCC---     CCT---        CCG---  CCG---  CCG---   −−−CGG      CCA---       CCG---        −−−AGG      −−−AGG        CCG---       −−−TGG     CCA---    CCC---  CCC---  CCC---        CCT---   CCA---    CCT---          CCG---                  −−−AGG         CCC---      CCA---
                                                               −−−TGG      −−−CGG                     −−−AGG                 −−−GGG  −−−GGG      CCC---    −−−CGG                              CCT---  CCC---       CCG---         −−−CGG  −−−GGG    CCG---                                      CCC---  −−−TGG             −−−GGG       CCC---  CCC---      −−−CGG            −−−AGG                −−−TGG                  −−−TGG  −−−GGG                  CCG---     CCG---         CCA---          CCC---          CCC---      CCC---     CCC---     −−−GGG    −−−AGG      −−−CGG                       −−−GGG            CCA---       −−−CGG    −−−TGG       CCC---  CCC---            −−−TGG                             −−−CGG                               CCC---        CCG---
                                                                                                                                   −−−TGG         CCA---                                                CCA---      −−−CGG                −−−GGG   −−−CGG  CCT---                                −−−AGG   −−−GGG             −−−GGG       CCC---  CCC---      −−−GGG                                  −−−GGG                                                                  CCT---                    CCA---          CCG---      CCT---   −−−CGG  −−−TGG   −−−AGG  −−−AGG          −−−TGG                    −−−TGG        −−−CGG                        CCG---  CCC---  CCT---                −−−AGG                                                             CCA---     −−−CGG
                                                                                                                                                   −−−AGG                                                                                                                                         −−−GGG        −−−AGG        −−−GGG       CCT---  CCT---      −−−GGG                                  −−−GGG                                                                                           −−−CGG               −−−CGG           −−−GGG      −−−GGG         −−−AGG                                  −−−GGG                                   −−−CGG     CCA---    CCG---              −−−GGG                                                                        −−−GGG
                                                                                                                                                                                                                                                                                                                                              CCC---               −−−TGG                               −−−GGG                                                                                                                                             −−−GGG                                                                                          −−−GGG              −−−CGG
                                                                                                                                                                                                                                                                                                                                               CCC---
                                                                                                                                                                                                                                                                                                                                                    −−−TGG

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Predicted guide sequences for PAMs
Ranked by default from highest to lowest specificity score (Hsu et al., Nat Biot 2013). Click on a column title to rank by a score.
If you use this website, please cite our paper in NAR 2018. Too much information? Look at the CRISPOR manual.

Download as Excel tables: Guides /  Guides, all scores /  Off-targets /  Saturating mutagenesis assistant
Position/
Strand
Guide Sequence + PAM
+ Restriction Enzymes
+ Variants
Only G- Only GG- Only A-
MIT Specificity Score CFD Spec. score Predicted Efficiency
Show all scores
Outcome Off-targets for
0-1-2-3-4 mismatches
+ next to PAM
Genome Browser links to matches sorted by CFD off-target score
exons only chr6 only
39 / rev CGCCACTCGGCGAACGCAAG TGG
.................... ...
 Cloning / PCR primers
99 99 48 52 72 80 0 - 0 - 0 - 0 - 4
0 - 0 - 0 - 0 - 0

4 off-targets
show all...
57 / fw ATCCACTTGCGTTCGCCGAG TGG
.................... ...
Enzymes: NmeAIII, AcyI
 Cloning / PCR primers
99 99 61 33 81 87 0 - 0 - 0 - 1 - 7
0 - 0 - 0 - 0 - 0

8 off-targets
show all...
228 / fw GTCGCCGTCGACACGGCGCT GGG
.................... ...
Enzymes: HinP1I, BfoI
 Cloning / PCR primers
97 98 39 53 60 88 0 - 0 - 0 - 1 - 11
0 - 0 - 0 - 0 - 0

12 off-targets
show all...
221 / fw CTCCGCGGTCGCCGTCGACA CGG
.................... ...
Enzymes: BfoI, HinP1I, Hpy166II, TaqI, BceAI, HincII, SalI, FblI
 Cloning / PCR primers
99 98 47 42 64 76 0 - 0 - 0 - 1 - 14
0 - 0 - 0 - 0 - 0

15 off-targets
show all...
229 / fw TCGCCGTCGACACGGCGCTG GGG
.................... ...
Enzymes: HinP1I, BfoI
 Cloning / PCR primers
97 99 63 70 65 85 0 - 0 - 0 - 1 - 14
0 - 0 - 0 - 1 - 0

15 off-targets
show all...
227 / fw GGTCGCCGTCGACACGGCGC TGG
.................... ...
High GC content
Enzymes: HinP1I, BceAI, BfoI
 Cloning / PCR primers
96 98 30 24 60 71 0 - 0 - 0 - 2 - 14
0 - 0 - 0 - 0 - 0

16 off-targets
show all...
203 / rev CGCCGTGTCGACGGCGACCG CGG
.................... ...
High GC content
Enzymes: BstDSI, SacII, BspFNI, MspA1I, BseDI, Bsh1285I, SsiI
 Cloning / PCR primers
96 99 52 67 51 66 0 - 0 - 0 - 1 - 16
0 - 0 - 0 - 0 - 0

17 off-targets
show all...
65 / rev AATCGTCATTACAGTACCGC TGG
.................... ...
Enzymes: AsuHPI, MspA1I, SsiI, MaeIII, Tsp45I
 Cloning / PCR primers
98 98 57 28 57 74 0 - 0 - 0 - 0 - 20
0 - 0 - 0 - 0 - 0

20 off-targets
show all...
732 / rev GCGTTCCCGGTGGCGCAGTC TGG
.................... ...
 Cloning / PCR primers
97 99 26 64 60 82 0 - 0 - 0 - 1 - 19
0 - 0 - 0 - 0 - 0

20 off-targets
show all...
69 / fw TCGCCGAGTGGCGTCACCAG CGG
.................... ...
Enzymes: Csp6I, LpnPI, MspA1I, MwoI
 Cloning / PCR primers
95 98 61 98 57 54 0 - 0 - 0 - 1 - 20
0 - 0 - 0 - 1 - 0

21 off-targets
show all...
494 / rev AAGTCGGAGGGCCGGCTCGC GGG
.................... ...
Enzymes: BspFNI, BspCNI, BstDEI, BslI, Bsp1286I, Eco24I, BstC8I
 Cloning / PCR primers
94 97 42 61 70 85 0 - 0 - 0 - 2 - 19
0 - 0 - 0 - 0 - 0

21 off-targets
show all...
742 / rev CTGGTACTCGGCGTTCCCGG TGG
.................... ...
Enzymes: MspI, HinP1I, LpnPI, StyD4I, NciI
 Cloning / PCR primers
96 97 65 74 52 64 0 - 0 - 0 - 2 - 19
0 - 0 - 0 - 0 - 0

21 off-targets
show all...
478 / rev TCGCGGGCTCAGGCAACCGT GGG
.................... ...
Enzymes: BstDSI, BaeGI, NlaIV, BseDI, BanI, Bsp1286I, HpyCH4III
 Cloning / PCR primers
96 98 46 74 68 85 0 - 0 - 0 - 1 - 23
0 - 0 - 0 - 0 - 0

24 off-targets
show all...
569 / fw GAGCCGGTCGATGGACTAGC AGG
.................... ...
Enzymes: BstC8I, BtsIMutI, BtsI, MaeI
 Cloning / PCR primers
95 96 51 67 62 79 0 - 0 - 0 - 3 - 21
0 - 0 - 0 - 0 - 0

24 off-targets
show all...
52 / rev GTACCGCTGGTGACGCCACT CGG
.................... ...
 Cloning / PCR primers
96 97 56 45 81 88 0 - 0 - 0 - 2 - 25
0 - 0 - 0 - 0 - 0

27 off-targets
show all...
731 / rev CGTTCCCGGTGGCGCAGTCT GGG
.................... ...
Enzymes: HpyCH4V
 Cloning / PCR primers
95 98 34 60 61 65 0 - 0 - 1 - 4 - 22
0 - 0 - 0 - 2 - 0

27 off-targets
show all...
212 / rev TAGCCCCAGCGCCGTGTCGA CGG
.................... ...
Enzymes: BceAI, Hpy166II, Hpy99I, HincII, TaqI, SalI, FblI
 Cloning / PCR primers
97 97 48 59 52 84 0 - 0 - 0 - 1 - 27
0 - 0 - 0 - 0 - 0

28 off-targets
show all...
197 / rev GTCGACGGCGACCGCGGAGA TGG
.................... ...
Enzymes: MwoI, BtsIMutI, BtsI
 Cloning / PCR primers
92 97 51 70 48 60 0 - 0 - 0 - 4 - 31
0 - 0 - 0 - 1 - 1

35 off-targets
show all...
190 / rev GCGACCGCGGAGATGGCAGT GGG
.................... ...
Enzymes: AluBI, BtsIMutI, BtsI, TspRI
 Cloning / PCR primers
92 97 56 64 63 75 0 - 0 - 0 - 3 - 34
0 - 0 - 0 - 1 - 0

37 off-targets
show all...
506 / rev CACGCTCCCGGGAAGTCGGA GGG
.................... ...
Enzymes: BshFI, MspI, Hpy188I, PspPI, LpnPI, NaeI, Cfr10I, BstC8I
 Cloning / PCR primers
92 95 54 56 65 78 0 - 0 - 0 - 6 - 33
0 - 0 - 0 - 0 - 0

39 off-targets
show all...
531 / fw CGACTTCCCGGGAGCGTGGC GGG
.................... ...
 Cloning / PCR primers
91 96 34 43 64 91 0 - 0 - 0 - 3 - 37
0 - 0 - 0 - 0 - 0

40 off-targets
show all...
510 / rev CCGCCACGCTCCCGGGAAGT CGG
.................... ...
Enzymes: Hpy188I
 Cloning / PCR primers
92 97 43 74 68 82 0 - 0 - 0 - 1 - 41
0 - 0 - 0 - 0 - 1

42 off-targets
show all...
479 / rev CTCGCGGGCTCAGGCAACCG TGG
.................... ...
Enzymes: Bsp1286I, BstDSI, BaeGI, HpyCH4III, BseDI
 Cloning / PCR primers
93 97 52 54 67 82 0 - 0 - 0 - 1 - 42
0 - 0 - 0 - 0 - 0

43 off-targets
show all...
597 / rev ATGGCCGCTCCAGCAGACCG TGG
.................... ...
Enzymes: BstDSI, BslI, HpyCH4III, BseDI
 Cloning / PCR primers
92 96 63 49 66 84 0 - 0 - 1 - 0 - 42
0 - 0 - 0 - 0 - 2

43 off-targets
show all...
93 / fw ACTGTAATGACGATTGCAGC AGG
.................... ...
Enzymes: EcoP15I, BstV1I, ApeKI, Fsp4HI, HpyCH4V
 Cloning / PCR primers
94 96 57 35 70 79 0 - 0 - 0 - 2 - 44
0 - 0 - 0 - 1 - 0

46 off-targets
show all...
477 / rev CGCGGGCTCAGGCAACCGTG GGG
.................... ...
Enzymes: BstDSI, BaeGI, NlaIV, BseDI, BanI, Bsp1286I, HpyCH4III
 Cloning / PCR primers
92 96 64 79 74 88 0 - 0 - 1 - 4 - 42
0 - 0 - 0 - 0 - 0

47 off-targets
show all...
608 / fw GGCGCCGAGCCACGGTCTGC TGG
.................... ...
High GC content
Enzymes: BpmI
 Cloning / PCR primers
87 94 34 15 62 80 0 - 0 - 1 - 5 - 41
0 - 0 - 0 - 0 - 1

47 off-targets
show all...
675 / rev GATTCGGTGTCTGAGCTACT AGG
.................... ...
Enzymes: MaeI
 Cloning / PCR primers
89 95 56 58 72 87 0 - 0 - 0 - 6 - 41
0 - 0 - 0 - 1 - 1

47 off-targets
show all...
507 / rev CCACGCTCCCGGGAAGTCGG AGG
.................... ...
Enzymes: MspI, BshFI, LpnPI, Hpy188I, PspPI
 Cloning / PCR primers
88 93 53 49 63 88 0 - 0 - 0 - 3 - 46
0 - 0 - 0 - 0 - 0

49 off-targets
show all...
532 / fw GACTTCCCGGGAGCGTGGCG GGG
.................... ...
Enzymes: MwoI
 Cloning / PCR primers
93 95 44 55 65 85 0 - 0 - 0 - 3 - 47
0 - 0 - 0 - 0 - 1

50 off-targets
show all...
495 / rev GAAGTCGGAGGGCCGGCTCG CGG
.................... ...
Enzymes: BspFNI, Bsp1286I, BslI, Eco24I, BstC8I
 Cloning / PCR primers
91 94 49 54 71 80 0 - 0 - 0 - 4 - 47
0 - 0 - 0 - 0 - 0

51 off-targets
show all...
745 / rev GTGCTGGTACTCGGCGTTCC CGG
.................... ...
Inefficient
Enzymes: MspI, NciI, LpnPI, StyD4I
 Cloning / PCR primers
95 97 39 72 55 72 0 - 0 - 0 - 0 - 54
0 - 0 - 0 - 0 - 0

54 off-targets
show all...
133 / rev TGTGCCGCACCGCCTCTGGG AGG
.................... ...
Enzymes: BstENI, BslI
 Cloning / PCR primers
90 94 62 51 67 83 0 - 0 - 0 - 8 - 48
0 - 0 - 0 - 2 - 1

56 off-targets
show all...
503 / fw GGTTGCCTGAGCCCGCGAGC CGG
.................... ...
Enzymes: BshFI, MnlI, MspI, PspPI, LpnPI, BslI, NaeI, Cfr10I, BstC8I
 Cloning / PCR primers
93 95 53 39 67 82 0 - 0 - 0 - 3 - 53
0 - 0 - 0 - 0 - 0

56 off-targets
show all...
530 / fw CCGACTTCCCGGGAGCGTGG CGG
.................... ...
 Cloning / PCR primers
90 92 56 66 67 85 0 - 0 - 0 - 3 - 54
0 - 0 - 0 - 0 - 2

57 off-targets
show all...
430 / rev CAGCGGGCTGCTCACGGCTT CGG
.................... ...
Inefficient
Enzymes: BceAI, Hpy188I
 Cloning / PCR primers
90 95 42 80 61 67 0 - 0 - 1 - 4 - 54
0 - 0 - 0 - 0 - 0

59 off-targets
show all...
697 / fw TAGTAGCTCAGACACCGAAT CGG
.................... ...
Enzymes: HinfI, Hpy188I, PfeI
 Cloning / PCR primers
94 95 63 57 55 72 0 - 0 - 0 - 5 - 54
0 - 0 - 0 - 0 - 0

59 off-targets
show all...
189 / rev CGACCGCGGAGATGGCAGTG GGG
.................... ...
Enzymes: AluBI, BtsIMutI, BtsI, TspRI
 Cloning / PCR primers
85 93 73 60 68 88 0 - 0 - 0 - 4 - 58
0 - 0 - 0 - 2 - 4

62 off-targets
show all...
236 / rev TAAGGCGACAAGGCAGGCGC CGG
.................... ...
Enzymes: NlaIV, AcyI, NciI, MspI, BfoI, HinP1I, BanI, LpnPI, SspDI, StyD4I, BstC8I
 Cloning / PCR primers
89 92 44 47 63 72 0 - 0 - 1 - 7 - 54
0 - 0 - 0 - 2 - 0

62 off-targets
show all...
552 / rev CTGCCTGCTAGTCCATCGAC CGG
.................... ...
Enzymes: NlaIV, MspI, BslI, TaqI, LpnPI, Bsh1285I, LmnI, Cfr10I
 Cloning / PCR primers
93 94 69 58 72 79 0 - 0 - 0 - 4 - 61
0 - 0 - 0 - 0 - 0

65 off-targets
show all...
96 / fw GTAATGACGATTGCAGCAGG AGG
.................... ...
Enzymes: EcoP15I, BtsCI
 Cloning / PCR primers
81 93 65 75 70 78 0 - 0 - 0 - 12 - 56
0 - 0 - 0 - 2 - 5

68 off-targets
show all...
613 / fw CGAGCCACGGTCTGCTGGAG CGG
.................... ...
Enzymes: BshFI, TauI, MwoI, BpmI, NlaIII, AcoI, Fsp4HI
 Cloning / PCR primers
79 91 49 62 68 77 0 - 0 - 1 - 10 - 58
0 - 0 - 0 - 3 - 10

69 off-targets
show all...
527 / fw CCTCCGACTTCCCGGGAGCG TGG
.................... ...
 Cloning / PCR primers
89 93 56 38 71 85 0 - 0 - 0 - 7 - 64
0 - 0 - 0 - 0 - 2

71 off-targets
show all...
592 / rev CGCTCCAGCAGACCGTGGCT CGG
.................... ...
Enzymes: BstDSI, NlaIV, AcyI, BfoI, HinP1I, BseDI, BanI, BslI, SspDI
 Cloning / PCR primers
89 93 43 44 66 75 0 - 0 - 3 - 3 - 65
0 - 0 - 0 - 0 - 0

71 off-targets
show all...
519 / fw GAGCCGGCCCTCCGACTTCC CGG
.................... ...
Inefficient
Enzymes: BmeT110I, MspI, BseDI, NciI, LpnPI, StyD4I, Cfr9I
 Cloning / PCR primers
90 92 39 51 77 85 0 - 0 - 1 - 5 - 67
0 - 0 - 0 - 0 - 0

73 off-targets
show all...
533 / fw ACTTCCCGGGAGCGTGGCGG GGG
.................... ...
Enzymes: BshFI, MwoI
 Cloning / PCR primers
91 94 46 81 62 79 0 - 0 - 0 - 3 - 71
0 - 0 - 0 - 0 - 1

74 off-targets
show all...
662 / fw CCCTCCGCGGACAGCCTCTC TGG
.................... ...
Inefficient
Enzymes: BshFI, BalI, MwoI, MnlI, BglI, AcoI
 Cloning / PCR primers
82 90 35 43 70 78 0 - 0 - 0 - 8 - 72
0 - 0 - 0 - 1 - 0

80 off-targets
show all...
237 / fw GACACGGCGCTGGGGCTACC CGG
.................... ...
Enzymes: NlaIV, AcyI, NciI, MspI, BfoI, HinP1I, BanI, LpnPI, SspDI, StyD4I
 Cloning / PCR primers
83 92 48 47 69 88 0 - 0 - 1 - 2 - 82
0 - 0 - 0 - 1 - 3

85 off-targets
show all...
488 / rev GAGGGCCGGCTCGCGGGCTC AGG
.................... ...
High GC content
Enzymes: Bsp1286I, BstDEI, BstC8I, Eco24I, BspCNI
 Cloning / PCR primers
81 93 36 80 64 73 0 - 0 - 1 - 8 - 76
0 - 0 - 0 - 2 - 0

85 off-targets
show all...
643 / rev GCCAGAGAGGCTGTCCGCGG AGG
.................... ...
Enzymes: BstDSI, BslFI, SacII, BspFNI, MspA1I, BseDI, BslI, SsiI
 Cloning / PCR primers
90 91 47 79 68 81 0 - 0 - 1 - 8 - 77
0 - 0 - 0 - 0 - 0

86 off-targets
show all...
361 / fw TCCGGGGATGGAGAGCCGAA AGG
.................... ...
Enzymes: NlaIII
 Cloning / PCR primers
86 91 63 61 55 83 0 - 0 - 0 - 3 - 87
0 - 0 - 0 - 0 - 1

90 off-targets
show all...
520 / fw AGCCGGCCCTCCGACTTCCC GGG
.................... ...
Enzymes: BmeT110I, MspI, BseDI, NciI, LpnPI, StyD4I, Cfr9I
 Cloning / PCR primers
85 92 51 45 75 82 0 - 0 - 1 - 6 - 85
0 - 0 - 1 - 1 - 0

92 off-targets
show all...
642 / rev CCAGAGAGGCTGTCCGCGGA GGG
.................... ...
Enzymes: BstDSI, BslFI, SacII, BspFNI, MspA1I, BseDI, BslI
 Cloning / PCR primers
91 96 47 63 68 76 0 - 0 - 0 - 5 - 87
0 - 0 - 0 - 0 - 0

92 off-targets
show all...
746 / fw TGCACCCCAGACTGCGCCAC CGG
.................... ...
Enzymes: MspI, NciI, LpnPI, StyD4I
 Cloning / PCR primers
90 97 44 63 57 63 0 - 0 - 1 - 4 - 88
0 - 0 - 0 - 0 - 0

93 off-targets
show all...
175 / fw AGAGGAGCGCTCGCATCACA AGG
.................... ...
Enzymes: AsuHPI, BstEII, DraIII, MaeIII, Tsp45I
 Cloning / PCR primers
88 90 61 51 64 78 0 - 0 - 0 - 8 - 86
0 - 0 - 0 - 0 - 0

94 off-targets
show all...
311 / fw TGGGATTAGGAGCTTACTTG GGG
.................... ...
Inefficient
 Cloning / PCR primers
84 94 53 44 64 74 0 - 0 - 0 - 12 - 83
0 - 0 - 0 - 3 - 0

95 off-targets
show all...
453 / rev ACCATCTTCTCTGGCGGTTC AGG
.................... ...
Inefficient
Enzymes: MwoI, ApeKI, Fsp4HI, BstV1I
 Cloning / PCR primers
85 94 29 53 54 60 0 - 0 - 1 - 7 - 88
0 - 0 - 0 - 1 - 1

96 off-targets
show all...
691 / rev TTCTTCATAGAAATCCGATT CGG
.................... ...
Enzymes: HinfI, Hpy188I, PfeI
 Cloning / PCR primers
90 94 45 43 54 81 0 - 0 - 1 - 8 - 90
0 - 0 - 0 - 1 - 0

99 off-targets
show all...
312 / fw GGGATTAGGAGCTTACTTGG GGG
.................... ...
 Cloning / PCR primers
84 88 61 67 67 84 0 - 0 - 0 - 9 - 94
0 - 0 - 0 - 0 - 2

103 off-targets
show all...
628 / fw TGGAGCGGCCATGCTTGGCC CGG
.................... ...
Inefficient
Enzymes: BshFI, BmeT110I, NlaIV, BslFI, MspI, PpuMI, EcoO109I, BseDI, PspPI, Bme18I, NciI, LpnPI, StyD4I, KflI, Cfr9I
 Cloning / PCR primers
84 88 36 62 64 82 0 - 0 - 0 - 11 - 92
0 - 0 - 0 - 1 - 5

103 off-targets
show all...
462 / rev CCGTGGGGCACCATCTTCTC TGG
.................... ...
Inefficient
 Cloning / PCR primers
76 93 39 57 66 67 0 - 0 - 2 - 22 - 81
0 - 0 - 0 - 2 - 2

105 off-targets
show all...
235 / rev AAGGCGACAAGGCAGGCGCC GGG
.................... ...
Enzymes: NlaIV, AcyI, NciI, MspI, BfoI, HinP1I, BanI, LpnPI, SspDI, StyD4I
 Cloning / PCR primers
80 90 52 70 68 70 0 - 0 - 1 - 12 - 93
0 - 0 - 0 - 0 - 0

106 off-targets
show all...
502 / rev CTCCCGGGAAGTCGGAGGGC CGG
.................... ...
Enzymes: BshFI, MspI, PspPI, LpnPI, NaeI, Cfr10I, BstC8I
 Cloning / PCR primers
83 88 50 51 69 76 0 - 0 - 0 - 15 - 91
0 - 0 - 0 - 0 - 1

106 off-targets
show all...
414 / fw TTGGCACTCTGGTTGGTAAG AGG
.................... ...
Enzymes: BspTNI, BcoDI
 Cloning / PCR primers
83 90 54 56 80 85 0 - 0 - 0 - 5 - 102
0 - 0 - 0 - 0 - 3

107 off-targets
show all...
636 / rev AGGCTGTCCGCGGAGGGGAC TGG
.................... ...
Enzymes: BmuI, Bse1I, Eam1105I, BslFI, BslI
 Cloning / PCR primers
79 88 30 59 69 76 0 - 0 - 2 - 6 - 99
0 - 0 - 0 - 0 - 1

107 off-targets
show all...
137 / rev CCTCTGTGCCGCACCGCCTC TGG
.................... ...
Enzymes: BstENI, BslI, MnlI
 Cloning / PCR primers
89 96 30 49 71 72 0 - 0 - 0 - 1 - 107
0 - 0 - 0 - 0 - 0

108 off-targets
show all...
635 / rev GGCTGTCCGCGGAGGGGACT GGG
.................... ...
Enzymes: Bse1I, NlaIV, Eam1105I, BslFI, PspPI, Bme18I, BmuI
 Cloning / PCR primers
74 88 25 58 67 77 0 - 0 - 0 - 14 - 97
0 - 0 - 0 - 0 - 1

111 off-targets
show all...
539 / rev CATCGACCGGCTCCTCCTCC TGG
.................... ...
Enzymes: BshFI, MnlI, LpnPI, BstNI, BslI, StyD4I
 Cloning / PCR primers
68 88 38 18 73 66 0 - 0 - 0 - 16 - 97
0 - 0 - 0 - 4 - 12

113 off-targets
show all...
649 / fw GGGACCCCCAGTCCCCTCCG CGG
.................... ...
High GC content
Enzymes: BstDSI, SacII, MnlI, BspFNI, MspA1I, BseDI, BslI, SsiI
 Cloning / PCR primers
79 88 68 70 68 81 0 - 0 - 1 - 9 - 104
0 - 0 - 0 - 1 - 1

114 off-targets
show all...
191 / rev GGCGACCGCGGAGATGGCAG TGG
.................... ...
Enzymes: MwoI, BtsIMutI, BtsI, TspRI
 Cloning / PCR primers
77 88 46 48 64 69 0 - 0 - 0 - 10 - 105
0 - 0 - 0 - 1 - 1

115 off-targets
show all...
634 / rev GCTGTCCGCGGAGGGGACTG GGG
.................... ...
Enzymes: Bse1I, NlaIV, Eam1105I, BslFI, EcoO109I, PpuMI, PspPI, Bme18I, BmuI, KflI
 Cloning / PCR primers
75 86 39 64 66 81 0 - 0 - 1 - 9 - 105
0 - 0 - 0 - 1 - 1

115 off-targets
show all...
289 / rev CCAAGTAAGCTCCTAATCCC AGG
.................... ...
Enzymes: NlaIV, BseDI, LpnPI, BstNI, LmnI, StyD4I
 Cloning / PCR primers
74 92 54 40 70 79 0 - 0 - 0 - 7 - 109
0 - 0 - 0 - 0 - 0

116 off-targets
show all...
136 / rev CTCTGTGCCGCACCGCCTCT GGG
.................... ...
Inefficient
Enzymes: BstENI, BslI, MnlI
 Cloning / PCR primers
88 90 35 40 68 79 0 - 0 - 0 - 4 - 113
0 - 0 - 0 - 0 - 0

117 off-targets
show all...
310 / fw CTGGGATTAGGAGCTTACTT GGG
.................... ...
Inefficient
 Cloning / PCR primers
81 89 38 43 65 82 0 - 0 - 0 - 9 - 109
0 - 0 - 0 - 1 - 3

118 off-targets
show all...
747 / fw GCACCCCAGACTGCGCCACC GGG
.................... ...
Enzymes: MspI, NciI, LpnPI, StyD4I
 Cloning / PCR primers
82 91 58 48 60 77 0 - 0 - 0 - 10 - 108
0 - 0 - 0 - 0 - 1

118 off-targets
show all...
633 / rev CTGTCCGCGGAGGGGACTGG GGG
.................... ...
Enzymes: Bse1I, NlaIV, Eam1105I, EcoO109I, PpuMI, PspPI, Bme18I, BmuI, KflI
 Cloning / PCR primers
77 86 44 67 65 78 0 - 0 - 0 - 12 - 107
0 - 0 - 0 - 6 - 5

119 off-targets
show all...
669 / fw CGGACAGCCTCTCTGGCCAA GGG
.................... ...
Enzymes: BshFI, ErhI, BalI, BseDI, AcoI
 Cloning / PCR primers
69 86 58 83 63 81 0 - 0 - 2 - 15 - 102
0 - 0 - 1 - 1 - 5

119 off-targets
show all...
436 / rev TTCAGGCAGCGGGCTGCTCA CGG
.................... ...
Enzymes: BceAI
 Cloning / PCR primers
76 85 52 51 82 88 0 - 0 - 0 - 16 - 107
0 - 0 - 0 - 0 - 2

123 off-targets
show all...
623 / fw TCTGCTGGAGCGGCCATGCT TGG
.................... ...
Enzymes: BshFI, MspI, PspPI, NciI, NlaIII, LpnPI, StyD4I
 Cloning / PCR primers
76 89 43 50 68 90 0 - 0 - 1 - 11 - 112
0 - 0 - 0 - 0 - 3

124 off-targets
show all...
329 / rev CCATCCCCGGACTCTCCAGA GGG
.................... ...
Enzymes: LpnPI, Hpy188III, BslI
 Cloning / PCR primers
72 86 50 36 73 80 0 - 0 - 1 - 21 - 103
0 - 0 - 0 - 3 - 2

125 off-targets
show all...
325 / rev CCCCGGACTCTCCAGAGGGG GGG
.................... ...
Enzymes: BslI
 Cloning / PCR primers
62 81 57 61 70 79 1 - 0 - 3 - 16 - 107
1 - 0 - 1 - 2 - 4

127 off-targets
show all...
407 / fw GGTCAGCTTGGCACTCTGGT TGG
.................... ...
 Cloning / PCR primers
71 88 46 35 70 69 0 - 0 - 2 - 22 - 103
0 - 0 - 0 - 0 - 4

127 off-targets
show all...
328 / rev CATCCCCGGACTCTCCAGAG GGG
.................... ...
Enzymes: LpnPI, Hpy188III, BslI
 Cloning / PCR primers
71 85 62 61 75 83 0 - 0 - 1 - 20 - 108
0 - 0 - 1 - 2 - 6

129 off-targets
show all...
629 / fw GGAGCGGCCATGCTTGGCCC GGG
.................... ...
Enzymes: BshFI, BmeT110I, NlaIV, BslFI, MspI, PpuMI, EcoO109I, BseDI, PspPI, Bme18I, NciI, LpnPI, StyD4I, KflI, Cfr9I
 Cloning / PCR primers
70 87 43 52 64 70 0 - 0 - 2 - 10 - 123
0 - 0 - 0 - 4 - 4

135 off-targets
show all...
665 / rev CTGAGCTACTAGGTTGCCCT TGG
.................... ...
Enzymes: BshFI, BalI, BseDI, ErhI, LpnPI, AcoI
 Cloning / PCR primers
81 88 51 50 65 78 0 - 0 - 2 - 13 - 122
0 - 0 - 0 - 1 - 0

137 off-targets
show all...
730 / rev GTTCCCGGTGGCGCAGTCTG GGG
.................... ...
Enzymes: BsgI, HpyCH4V
 Cloning / PCR primers
75 83 58 75 57 73 0 - 0 - 1 - 6 - 130
0 - 0 - 0 - 0 - 0

137 off-targets
show all...
718 / fw GGATTTCTATGAAGAAATCG AGG
.................... ...
Enzymes: AsuHPI, TaqI
 Cloning / PCR primers
73 83 68 17 56 74 0 - 0 - 1 - 16 - 121
0 - 0 - 0 - 1 - 2

138 off-targets
show all...
415 / fw TGGCACTCTGGTTGGTAAGA GGG
.................... ...
Enzymes: BspTNI, BcoDI
 Cloning / PCR primers
78 84 45 56 77 79 0 - 0 - 1 - 13 - 125
0 - 0 - 0 - 2 - 3

139 off-targets
show all...
345 / fw CCCCCCCTCTGGAGAGTCCG GGG
.................... ...
Enzymes: PpsI, MspI, BtsCI, BseDI, NciI, LpnPI, StyD4I, HinfI
 Cloning / PCR primers
70 83 56 59 64 74 0 - 0 - 3 - 10 - 128
0 - 0 - 2 - 1 - 0

141 off-targets
show all...
616 / rev TGGGGGTCCCGGGCCAAGCA TGG
.................... ...
Enzymes: BshFI, TauI, NlaIII, AcoI, SsiI, Fsp4HI
 Cloning / PCR primers
78 87 45 53 72 89 0 - 0 - 0 - 13 - 128
0 - 0 - 0 - 1 - 0

141 off-targets
show all...
309 / fw CCTGGGATTAGGAGCTTACT TGG
.................... ...
 Cloning / PCR primers
84 88 28 40 64 85 0 - 0 - 0 - 11 - 131
0 - 0 - 0 - 0 - 1

142 off-targets
show all...
157 / fw CCAGAGGCGGTGCGGCACAG AGG
.................... ...
Enzymes: HinP1I, BseRI
 Cloning / PCR primers
80 88 59 72 70 88 0 - 0 - 0 - 10 - 133
0 - 0 - 0 - 0 - 0

143 off-targets
show all...
646 / rev TTGGCCAGAGAGGCTGTCCG CGG
.................... ...
Enzymes: BstDSI, SacII, BspFNI, MspA1I, BseDI, BslI, SsiI
 Cloning / PCR primers
77 86 63 70 68 87 0 - 0 - 0 - 17 - 130
0 - 0 - 0 - 0 - 2

147 off-targets
show all...
656 / rev TAGGTTGCCCTTGGCCAGAG AGG
.................... ...
Enzymes: LpnPI, MwoI, BglI
 Cloning / PCR primers
67 79 58 88 72 84 0 - 0 - 2 - 17 - 130
0 - 0 - 1 - 2 - 14

149 off-targets
show all...
518 / rev GGCCTCCCCCGCCACGCTCC CGG
.................... ...
High GC content
Enzymes: BmeT110I, MspI, BseDI, NciI, LpnPI, StyD4I, LmnI, Cfr9I
 Cloning / PCR primers
78 88 32 37 71 77 0 - 0 - 3 - 13 - 134
0 - 0 - 0 - 2 - 2

150 off-targets
show all...
582 / fw GACTAGCAGGCAGTGCTGCA GGG
.................... ...
Enzymes: PstI, BfmI, ApeKI, Fsp4HI, HpyCH4V
 Cloning / PCR primers
68 88 54 45 64 89 0 - 0 - 3 - 21 - 126
0 - 0 - 0 - 4 - 4

150 off-targets
show all...
403 / fw TGGGGGTCAGCTTGGCACTC TGG
.................... ...
 Cloning / PCR primers
73 86 41 59 69 72 0 - 0 - 3 - 13 - 139
0 - 0 - 0 - 1 - 2

155 off-targets
show all...
330 / rev TCCATCCCCGGACTCTCCAG AGG
.................... ...
Enzymes: LpnPI, Hpy188III, BslI
 Cloning / PCR primers
66 82 51 60 73 74 0 - 0 - 3 - 23 - 132
0 - 0 - 0 - 1 - 2

158 off-targets
show all...
144 / fw ATGGGGCTTCCTCCCAGAGG CGG
.................... ...
Enzymes: BstENI, BslI
 Cloning / PCR primers
66 79 63 83 64 81 0 - 0 - 7 - 24 - 132
0 - 0 - 3 - 6 - 8

163 off-targets
show all...
242 / rev AGGAGCTAAGGCGACAAGGC AGG
.................... ...
Enzymes: NlaIV, AcyI, BfoI, HinP1I, BanI, SspDI, BstC8I
 Cloning / PCR primers
80 88 49 42 64 72 0 - 0 - 1 - 13 - 149
0 - 0 - 0 - 0 - 0

163 off-targets
show all...
447 / rev TTCTCTGGCGGTTCAGGCAG CGG
.................... ...
Enzymes: MwoI, MspA1I, ApeKI, BstV1I, Fsp4HI, BstC8I
 Cloning / PCR primers
74 81 39 54 67 82 0 - 0 - 1 - 15 - 147
0 - 0 - 0 - 1 - 0

163 off-targets
show all...
141 / fw GCAATGGGGCTTCCTCCCAG AGG
.................... ...
Enzymes: BstENI, LpnPI, BslI
 Cloning / PCR primers
68 83 68 69 69 86 0 - 0 - 2 - 18 - 150
0 - 0 - 1 - 6 - 7

170 off-targets
show all...
344 / fw TCCCCCCCTCTGGAGAGTCC GGG
.................... ...
Enzymes: PpsI, MspI, BtsCI, BseDI, NciI, LpnPI, StyD4I, HinfI
 Cloning / PCR primers
65 79 43 40 59 65 0 - 0 - 1 - 22 - 154
0 - 0 - 0 - 6 - 8

177 off-targets
show all...
472 / fw GCCTGAACCGCCAGAGAAGA TGG
.................... ...
Enzymes: Bsp1286I, BanI, BaeGI, NlaIV, MboII
 Cloning / PCR primers
60 82 46 45 65 63 0 - 0 - 1 - 30 - 148
0 - 0 - 0 - 3 - 0

179 off-targets
show all...
343 / fw CTCCCCCCCTCTGGAGAGTC CGG
.................... ...
Enzymes: PpsI, MspI, BtsCI, BseDI, NciI, LpnPI, StyD4I, HinfI
 Cloning / PCR primers
69 82 32 59 59 70 0 - 0 - 4 - 24 - 152
0 - 0 - 0 - 2 - 4

180 off-targets
show all...
446 / rev TCTCTGGCGGTTCAGGCAGC GGG
.................... ...
Enzymes: MspA1I, BstC8I, ApeKI, Fsp4HI, BstV1I
 Cloning / PCR primers
65 85 41 63 73 68 0 - 0 - 0 - 15 - 169
0 - 0 - 0 - 1 - 1

184 off-targets
show all...
322 / rev CGGACTCTCCAGAGGGGGGG AGG
.................... ...
Enzymes: BslI
 Cloning / PCR primers
63 73 30 78 67 73 0 - 0 - 1 - 10 - 175
0 - 0 - 0 - 0 - 5

186 off-targets
show all...
600 / fw CAGGGCTGGGCGCCGAGCCA CGG
.................... ...
High GC content
Enzymes: BstDSI, BslI, HpyCH4III, BseDI
 Cloning / PCR primers
82 87 42 70 66 76 0 - 0 - 0 - 16 - 170
0 - 0 - 0 - 0 - 0

186 off-targets
show all...
326 / rev TCCCCGGACTCTCCAGAGGG GGG
.................... ...
Enzymes: BslI
 Cloning / PCR primers
67 72 51 67 72 79 0 - 0 - 1 - 26 - 161
0 - 0 - 0 - 3 - 8

188 off-targets
show all...
149 / fw GCTTCCTCCCAGAGGCGGTG CGG
.................... ...
Enzymes: TauI, Fsp4HI, BslI
 Cloning / PCR primers
76 80 61 61 71 85 0 - 0 - 3 - 24 - 165
0 - 0 - 0 - 2 - 1

192 off-targets
show all...
327 / rev ATCCCCGGACTCTCCAGAGG GGG
.................... ...
Enzymes: Hpy188III, BslI
 Cloning / PCR primers
59 63 52 60 74 83 0 - 0 - 4 - 16 - 176
0 - 0 - 4 - 6 - 3

196 off-targets
show all...
384 / fw ACATGGTTATGTTTCTGGAT GGG
.................... ...
Enzymes: Hpy188III, BtsCI
 Cloning / PCR primers
69 80 42 25 65 77 0 - 0 - 4 - 18 - 179
0 - 0 - 2 - 2 - 3

201 off-targets
show all...
342 / rev TCCTTTCGGCTCTCCATCCC CGG
.................... ...
Enzymes: MspI, BseDI, NciI, LpnPI, StyD4I, HinfI, BccI
 Cloning / PCR primers
63 82 45 40 64 66 0 - 0 - 0 - 27 - 182
0 - 0 - 0 - 4 - 6

209 off-targets
show all...
383 / fw GACATGGTTATGTTTCTGGA TGG
.................... ...
Enzymes: Hpy188III, BtsCI
 Cloning / PCR primers
65 74 47 33 67 80 0 - 0 - 1 - 34 - 174
0 - 0 - 0 - 3 - 4

209 off-targets
show all...
206 / fw GCTCCCCACTGCCATCTCCG CGG
.................... ...
Enzymes: BstDSI, MwoI, SacII, BspFNI, MspA1I, BseDI, Bsh1285I, SsiI
 Cloning / PCR primers
67 81 70 37 51 64 0 - 0 - 1 - 17 - 193
0 - 0 - 0 - 0 - 2

211 off-targets
show all...
395 / fw TTTCTGGATGGGGGTCAGCT TGG
.................... ...
Enzymes: AluBI
 Cloning / PCR primers
63 81 43 43 64 80 0 - 0 - 9 - 50 - 159
0 - 0 - 2 - 5 - 5

218 off-targets
show all...
246 / rev GAAGAGGAGCTAAGGCGACA AGG
.................... ...
Enzymes: BstC8I
 Cloning / PCR primers
80 82 61 37 65 81 0 - 0 - 1 - 17 - 201
0 - 0 - 0 - 2 - 1

219 off-targets
show all...
482 / fw CCAGAGAAGATGGTGCCCCA CGG
.................... ...
Enzymes: Bsp1286I, BstDSI, BaeGI, HpyCH4III, BseDI
 Cloning / PCR primers
55 79 69 64 67 90 0 - 1 - 17 - 30 - 184
0 - 0 - 2 - 3 - 1

232 off-targets
show all...
581 / fw GGACTAGCAGGCAGTGCTGC AGG
.................... ...
Enzymes: HpyCH4V, PstI, ApeKI, TspRI, BfmI, Fsp4HI
 Cloning / PCR primers
56 80 38 15 64 48 0 - 0 - 4 - 32 - 196
0 - 0 - 0 - 2 - 14

232 off-targets
show all...
280 / rev CTCCTAATCCCAGGCTCCCT GGG
.................... ...
Enzymes: NlaIV, NdeII, BspPI, BseDI, MflI, LpnPI, BstNI, LmnI, StyD4I, PasI
 Cloning / PCR primers
57 77 49 40 66 73 0 - 0 - 6 - 32 - 198
0 - 0 - 1 - 9 - 6

236 off-targets
show all...
386 / fw ATGGTTATGTTTCTGGATGG GGG
.................... ...
Enzymes: BtsCI
 Cloning / PCR primers
63 80 57 78 51 80 0 - 0 - 1 - 37 - 202
0 - 0 - 0 - 9 - 4

240 off-targets
show all...
367 / fw GATGGAGAGCCGAAAGGACA TGG
.................... ...
Enzymes: NlaIII, SmiMI
 Cloning / PCR primers
64 80 56 50 52 74 0 - 0 - 0 - 23 - 219
0 - 0 - 0 - 2 - 0

242 off-targets
show all...
321 / rev GGACTCTCCAGAGGGGGGGA GGG
.................... ...
Enzymes: BslI
 Cloning / PCR primers
65 66 46 54 65 74 0 - 0 - 4 - 22 - 231
0 - 0 - 0 - 0 - 3

257 off-targets
show all...
298 / fw ATCCCAGGGAGCCTGGGATT AGG
.................... ...
Enzymes: AluBI
 Cloning / PCR primers
61 78 39 54 62 71 0 - 0 - 2 - 34 - 222
0 - 0 - 0 - 4 - 8

258 off-targets
show all...
356 / rev GAAACATAACCATGTCCTTT CGG
.................... ...
Inefficient
 Cloning / PCR primers
60 82 39 20 52 74 0 - 0 - 6 - 35 - 221
0 - 0 - 1 - 3 - 7

262 off-targets
show all...
379 / fw AAAGGACATGGTTATGTTTC TGG
.................... ...
Inefficient
Enzymes: Hpy188III, BtsCI
 Cloning / PCR primers
67 79 21 36 55 67 0 - 0 - 3 - 32 - 234
0 - 0 - 0 - 2 - 2

269 off-targets
show all...
517 / rev GCCTCCCCCGCCACGCTCCC GGG
.................... ...
High GC content
Enzymes: BmeT110I, MspI, BseDI, NciI, LpnPI, StyD4I, LmnI, Cfr9I
 Cloning / PCR primers
64 81 39 46 75 85 0 - 0 - 4 - 24 - 244
0 - 0 - 0 - 0 - 3

272 off-targets
show all...
281 / rev GCTCCTAATCCCAGGCTCCC TGG
.................... ...
Enzymes: NlaIV, NdeII, BspPI, BseDI, MflI, LpnPI, BstNI, LmnI, StyD4I, PasI
 Cloning / PCR primers
61 72 42 9 66 85 0 - 0 - 5 - 23 - 246
0 - 0 - 1 - 6 - 13

274 off-targets
show all...
292 / fw GCCAAGATCCCAGGGAGCCT GGG
.................... ...
Inefficient
Enzymes: StyD4I, LpnPI, NlaIV, BstNI, BseDI
 Cloning / PCR primers
55 75 52 31 70 81 0 - 0 - 4 - 30 - 244
0 - 0 - 1 - 7 - 9

278 off-targets
show all...
262 / rev CTGGGATCTTGGCTGAGAAG AGG
.................... ...
Enzymes: AluBI, BseRI, MboII
 Cloning / PCR primers
60 75 52 64 70 73 0 - 0 - 7 - 33 - 251
0 - 0 - 2 - 4 - 2

291 off-targets
show all...
668 / fw GCGGACAGCCTCTCTGGCCA AGG
.................... ...
Enzymes: BshFI, BalI, MwoI, BseDI, BglI, ErhI, AcoI
 Cloning / PCR primers
61 83 44 39 65 74 0 - 0 - 4 - 22 - 265
0 - 0 - 0 - 5 - 3

291 off-targets
show all...
626 / rev CGGAGGGGACTGGGGGTCCC GGG
.................... ...
High GC content
Enzymes: BshFI, BmeT110I, NlaIV, Eam1105I, MspI, PpuMI, EcoO109I, BseDI, PspPI, Bme18I, NciI, LpnPI, StyD4I, KflI, Cfr9I
 Cloning / PCR primers
54 69 39 80 64 76 0 - 0 - 0 - 24 - 268
0 - 0 - 0 - 4 - 10

292 off-targets
show all...
283 / fw CTCTTCTCAGCCAAGATCCC AGG
.................... ...
Enzymes: NlaIV, NdeII, MflI, BseDI, LpnPI, BstNI, StyD4I, PasI
 Cloning / PCR primers
63 76 43 31 66 81 0 - 0 - 3 - 17 - 285
0 - 0 - 0 - 1 - 2

305 off-targets
show all...
126 / fw CTTAGAAAGAAGAGGGCAAT GGG
.................... ...
Enzymes: BseMI
 Cloning / PCR primers
57 77 43 52 66 76 0 - 0 - 6 - 38 - 270
0 - 0 - 0 - 7 - 2

314 off-targets
show all...
560 / fw CAGGAGGAGGAGCCGGTCGA TGG
.................... ...
Enzymes: Bsh1285I, BslI, TaqI, MaeI
 Cloning / PCR primers
86 87 50 65 81 87 0 - 0 - 0 - 29 - 298
0 - 0 - 0 - 3 - 0

327 off-targets
show all...
3 / rev TCTGCCACAGTGAAGATGAA AGG
.................... ...
Enzymes: TspDTI
 Cloning / PCR primers
57 75 57 32 72 82 0 - 1 - 5 - 40 - 282
0 - 0 - 1 - 4 - 8

328 off-targets
show all...
284 / fw TCTTCTCAGCCAAGATCCCA GGG
.................... ...
Enzymes: NlaIV, MflI, BseDI, LpnPI, BstNI, StyD4I, PasI
 Cloning / PCR primers
52 76 63 35 66 84 0 - 1 - 6 - 38 - 286
0 - 0 - 1 - 1 - 1

331 off-targets
show all...
349 / fw CCCTCTGGAGAGTCCGGGGA TGG
.................... ...
Enzymes: NciI, BtsCI, BseDI, StyD4I
 Cloning / PCR primers
72 83 46 64 63 57 0 - 0 - 4 - 11 - 317
0 - 0 - 1 - 0 - 2

332 off-targets
show all...
118 / fw GATGACAGCTTAGAAAGAAG AGG
.................... ...
Enzymes: MboII
 Cloning / PCR primers
50 68 61 30 72 71 0 - 0 - 6 - 33 - 297
0 - 0 - 1 - 5 - 13

336 off-targets
show all...
320 / rev GACTCTCCAGAGGGGGGGAG GGG
.................... ...
 Cloning / PCR primers
52 63 37 65 68 82 0 - 0 - 5 - 29 - 313
0 - 0 - 0 - 3 - 8

347 off-targets
show all...
254 / rev TTGGCTGAGAAGAGGAGCTA AGG
.................... ...
Enzymes: AluBI, BstDEI
 Cloning / PCR primers
50 71 55 56 71 84 0 - 0 - 5 - 49 - 301
0 - 0 - 1 - 9 - 5

355 off-targets
show all...
291 / fw AGCCAAGATCCCAGGGAGCC TGG
.................... ...
Inefficient
Enzymes: StyD4I, LpnPI, NlaIV, BstNI, BseDI
 Cloning / PCR primers
51 70 41 16 66 81 0 - 0 - 2 - 43 - 315
0 - 0 - 0 - 7 - 10

360 off-targets
show all...
627 / rev GCGGAGGGGACTGGGGGTCC CGG
.................... ...
High GC content
Enzymes: BshFI, BmeT110I, NlaIV, Eam1105I, MspI, PpuMI, EcoO109I, BseDI, PspPI, Bme18I, NciI, LpnPI, StyD4I, KflI, Cfr9I
 Cloning / PCR primers
34 77 34 85 63 71 0 - 1 - 2 - 25 - 332
0 - 1 - 1 - 4 - 4

360 off-targets
show all...
273 / rev TCCCAGGCTCCCTGGGATCT TGG
.................... ...
Enzymes: BspPI, BstDEI, MflI, NdeII
 Cloning / PCR primers
54 72 34 43 64 81 0 - 0 - 4 - 46 - 325
0 - 0 - 1 - 4 - 16

375 off-targets
show all...
385 / fw CATGGTTATGTTTCTGGATG GGG
.................... ...
Enzymes: Hpy188III, BtsCI
 Cloning / PCR primers
46 71 37 54 58 76 0 - 0 - 3 - 30 - 369
0 - 0 - 0 - 1 - 5

402 off-targets
show all...
587 / fw GCAGGCAGTGCTGCAGGGCT GGG
.................... ...
Enzymes: NlaIV, AcyI, BfoI, HinP1I, BanI, SspDI
 Cloning / PCR primers
52 70 30 72 60 85 0 - 0 - 1 - 51 - 354
0 - 0 - 1 - 3 - 12

406 off-targets
show all...
544 / fw GCGTGGCGGGGGAGGCCAGG AGG
.................... ...
High GC content
Enzymes: StyD4I, BseRI, LpnPI, BstNI, BslI
 Cloning / PCR primers
47 65 43 83 62 88 0 - 0 - 6 - 46 - 427
0 - 0 - 3 - 9 - 8

479 off-targets
show all...
536 / fw TCCCGGGAGCGTGGCGGGGG AGG
.................... ...
High GC content
Enzymes: BshFI, StyD4I, LpnPI, MwoI, BstNI
 Cloning / PCR primers
33 70 41 79 64 71 0 - 1 - 2 - 38 - 439
0 - 0 - 0 - 2 - 5

480 off-targets
show all...
119 / fw ATGACAGCTTAGAAAGAAGA GGG
.................... ...
Enzymes: BseMI, MboII
 Cloning / PCR primers
36 63 59 59 70 58 0 - 2 - 7 - 81 - 397
0 - 0 - 1 - 10 - 17

487 off-targets
show all...
586 / fw AGCAGGCAGTGCTGCAGGGC TGG
.................... ...
Enzymes: NlaIV, AcyI, BfoI, HinP1I, BanI, SspDI
 Cloning / PCR primers
45 68 35 37 62 82 0 - 1 - 9 - 52 - 431
0 - 0 - 1 - 4 - 6

493 off-targets
show all...
181 / rev GAGATGGCAGTGGGGAGCTA GGG
.................... ...
Enzymes: AluBI, Tsp45I, BstEII, MaeIII, MaeI
 Cloning / PCR primers
46 66 54 68 66 76 0 - 0 - 11 - 44 - 439
0 - 0 - 3 - 4 - 7

494 off-targets
show all...
125 / fw GCTTAGAAAGAAGAGGGCAA TGG
.................... ...
Enzymes: BseMI
 Cloning / PCR primers
45 61 55 32 66 78 0 - 0 - 5 - 50 - 442
0 - 0 - 0 - 6 - 18

497 off-targets
show all...
541 / fw GGAGCGTGGCGGGGGAGGCC AGG
.................... ...
Inefficient
High GC content
Enzymes: BshFI, MwoI, BseRI, LpnPI, BstNI, StyD4I
 Cloning / PCR primers
46 73 37 59 73 80 0 - 0 - 2 - 44 - 468
0 - 0 - 0 - 5 - 9

514 off-targets
More than 500 off-targets, showing 13 with score >1.0 show all...
334 / fw GTTTTCCCCCTCCCCCCCTC TGG
.................... ...
Not with U6/U3
Enzymes: BpmI, BslI, Hpy188III, MnlI
 Cloning / PCR primers
36 59 36 44 68 72 0 - 1 - 7 - 67 - 471
0 - 0 - 2 - 3 - 12

546 off-targets
More than 500 off-targets, showing 23 with score >1.0 show all...
127 / fw TTAGAAAGAAGAGGGCAATG GGG
.................... ...
Enzymes: BseMI
 Cloning / PCR primers
51 53 73 42 68 92 0 - 0 - 4 - 64 - 490
0 - 0 - 0 - 4 - 7

558 off-targets
More than 500 off-targets, showing 7 with score >1.0 show all...
319 / rev ACTCTCCAGAGGGGGGGAGG GGG
.................... ...
 Cloning / PCR primers
38 49 42 75 67 82 0 - 0 - 6 - 60 - 542
0 - 0 - 0 - 6 - 7

608 off-targets
More than 500 off-targets, showing 17 with score >1.0 show all...
182 / rev GGAGATGGCAGTGGGGAGCT AGG
.................... ...
Enzymes: AluBI, Tsp45I, MaeIII, MaeI
 Cloning / PCR primers
26 51 39 57 66 79 0 - 3 - 12 - 78 - 964
0 - 0 - 3 - 7 - 13

1057 off-targets
More than 500 off-targets, showing 23 with score >1.0 show all...
547 / fw TGGCGGGGGAGGCCAGGAGG AGG
.................... ...
High GC content
Enzymes: BseRI, NlaIV, BslI
 Cloning / PCR primers
19 33 37 95 57 74 0 - 0 - 6 - 158 - 1190
0 - 0 - 1 - 18 - 30

1354 off-targets
More than 500 off-targets, showing 12 with score >2.0 show all...
459 / rev TGGGGCACCATCTTCTCTGG CGG
.................... ...
 Cloning / PCR primers
24 72 66 69 60 75 0 - 0 - 1241 - 21 - 138
0 - 0 - 1 - 3 - 1

1400 off-targets
More than 500 off-targets, showing 24 with score >0.5 show all...
641 / rev CAGAGAGGCTGTCCGCGGAG GGG
.................... ...
Enzymes: BstDSI, Bse1I, BslFI, SacII, MspA1I, BseDI, BslI
 Cloning / PCR primers
16 32 59 83 70 72 0 - 0 - 0 - 1338 - 568
0 - 0 - 0 - 0 - 1

1906 off-targets
More than 500 off-targets, showing 4 with score >0.5 show all...
553 / fw GGGAGGCCAGGAGGAGGAGC CGG
.................... ...
Enzymes: NlaIV, MspI, TaqI, BseRI, LpnPI, Bsh1285I, Cfr10I
 Cloning / PCR primers
5 15 48 71 66 56 0 - 2 - 35 - 378 - 4403
0 - 0 - 9 - 72 - 1098

4818 off-targets
More than 500 off-targets, showing 20 with score >2.0 show all...
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CRISPR/Cas9 Guide Designer for chordate vertebrate ecdysozoans lophotrochozoans protostomes spongi corals plants butterflies metazoans genomes fruitflies insects nematodes mammals.