When reading the list of guide sequences and off-targets below, bear in mind that in case that the input sequence is really in the genome and just has a few differences, the software will use the first found match as the on-target as it cannot distinguish 0-mismatch off-targets from 0-mismatch on-targets. In this case, the specificity scores of guide sequences are too low. In other words, some guides may be fine, the problem may just be that the on-target is shown as an off-target.
Because there is no flanking sequence available, the guides in your sequence that are within 50bp of the ends will have no efficiency scores. The efficiency scores will instead be shown as '--'. Include more flanking sequence > 50bp to obtain these scores.
Input sequence not in genome, cannot show genome variants.
Position 0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 Sequence ACGCTGCCTGTCCGAGCCTAGTGCCGCCTTCCTCCCGCATCCGGCGCGGACGCCATGGTGCGGTTCAAGCACGGGTAAGCAGCGCTGTCCCCGCGCCGTCCCCGCGACCCGGGCCGCGCTCACCCCGCTGTCGCCGCAGGTACCTTC CCT--- CCT--- CCT--- CCG--- −−−CGG CCA--- −−−CGG CCC--- CCC--- CCC--- CCC--- −−−AGG CCG--- CCG--- CCC--- −−−TGG −−−GGG CCC--- CCC--- CCG--- CCC--- CCT--- CCG--- −−−CGG CCG--- CCG--- CCG--- CCG--- −−−CGG CCG--- −−−CGG −−−GGG
Position/ Strand |
Guide Sequence + PAM + Restriction Enzymes 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 No match, no chrom filter |
||
---|---|---|---|---|---|---|---|---|---|
34 / rev |
ATGGCGTCCGCGCCGGATGC GGG
Enzymes: BslI, MwoI, BtsCI
Cloning / PCR primers
|
98 | 99 | -- | -- | -- | -- |
0 - 0 - 0 - 2 - 7
0 - 0 - 0 - 0 - 0 9 off-targets |
show all... |
7 / rev |
AGGCGGCACTAGGCTCGGAC AGG
Enzymes: ApeKI, Hpy188I, Fsp4HI, BstV1I
Cloning / PCR primers
|
96 | 97 | -- | -- | -- | -- |
0 - 0 - 0 - 1 - 25
0 - 0 - 0 - 0 - 0 26 off-targets |
show all... |
35 / rev |
CATGGCGTCCGCGCCGGATG CGG
Enzymes: BslI, MwoI, BtsCI
Cloning / PCR primers
|
96 | 99 | -- | -- | -- | -- |
0 - 0 - 0 - 1 - 16
0 - 0 - 0 - 0 - 0 17 off-targets |
show all... |
41 / rev |
CCGCACCATGGCGTCCGCGC CGG
High GC content Enzymes: MwoI, BtsCI, MspI, HinP1I, BspFNI, LpnPI, SsiI
Cloning / PCR primers
|
95 | 98 | -- | -- | -- | -- |
0 - 0 - 0 - 4 - 26
0 - 0 - 0 - 0 - 1 30 off-targets |
show all... |
56 / fw |
CGCATCCGGCGCGGACGCCA TGG
High GC content Enzymes: BstDSI, AcyI, BseDI, ErhI, HgaI, BslI, NlaIII, Bsp19I
Cloning / PCR primers
|
95 | 98 | 45 | 44 | 74 | 80 |
0 - 0 - 0 - 1 - 15
0 - 0 - 0 - 0 - 1 16 off-targets |
show all... |
124 / rev |
AAGGTACCTGCGGCGACAGC GGG
Enzymes: AsuHPI, MspA1I, MwoI
Cloning / PCR primers
|
95 | 97 | -- | -- | -- | -- |
0 - 0 - 0 - 1 - 40
0 - 0 - 0 - 0 - 0 41 off-targets |
show all... |
47 / fw |
CCTTCCTCCCGCATCCGGCG CGG
Enzymes: AcyI, MwoI, MspI, HinP1I, HgaI, LpnPI, BslI, BspFNI
Cloning / PCR primers
|
94 | 96 | -- | -- | -- | -- |
0 - 0 - 0 - 2 - 50
0 - 0 - 0 - 0 - 0 52 off-targets |
show all... |
138 / fw |
GCTCACCCCGCTGTCGCCGC AGG
High GC content Enzymes: TauI, MwoI, Csp6I, BanI, Asp718I, NlaIV, SsiI, Fsp4HI
Cloning / PCR primers
|
94 | 96 | -- | -- | -- | -- |
0 - 0 - 0 - 7 - 49
0 - 0 - 0 - 0 - 0 56 off-targets |
show all... |
31 / rev |
GCGTCCGCGCCGGATGCGGG AGG
High GC content Enzymes: BslI
Cloning / PCR primers
|
92 | 97 | -- | -- | -- | -- |
0 - 0 - 0 - 4 - 47
0 - 0 - 0 - 1 - 0 51 off-targets |
show all... |
123 / rev |
AGGTACCTGCGGCGACAGCG GGG
Enzymes: AsuHPI, MspA1I, MwoI
Cloning / PCR primers
|
92 | 96 | -- | -- | -- | -- |
0 - 0 - 0 - 4 - 35
0 - 0 - 0 - 1 - 0 39 off-targets |
show all... |
100 / rev |
TGAGCGCGGCCCGGGTCGCG GGG
High GC content Enzymes: BspFNI, BceAI, MwoI, BslFI, BslI
Cloning / PCR primers
|
90 | 94 | -- | -- | -- | -- |
0 - 0 - 0 - 5 - 56
0 - 0 - 0 - 0 - 1 61 off-targets |
show all... |
61 / fw |
CCGGCGCGGACGCCATGGTG CGG
High GC content Enzymes: BstDSI, ErhI, BseDI, AgsI, BslI, Bsp19I
Cloning / PCR primers
|
89 | 95 | 59 | 86 | 73 | 79 |
0 - 0 - 0 - 1 - 40
0 - 0 - 0 - 0 - 0 41 off-targets |
show all... |
125 / rev |
GAAGGTACCTGCGGCGACAG CGG
Enzymes: AsuHPI, MspA1I, MwoI
Cloning / PCR primers
|
89 | 94 | -- | -- | -- | -- |
0 - 0 - 0 - 2 - 69
0 - 0 - 0 - 0 - 0 71 off-targets |
show all... |
109 / rev |
ACAGCGGGGTGAGCGCGGCC CGG
Inefficient High GC content Enzymes: BshFI, BmeT110I, TauI, MwoI, MspI, BseDI, PspPI, NciI, LpnPI, StyD4I, Cfr9I, Fsp4HI
Cloning / PCR primers
|
87 | 93 | -- | -- | -- | -- |
0 - 0 - 0 - 8 - 67
0 - 0 - 0 - 0 - 2 75 off-targets |
show all... |
27 / rev |
CCGCGCCGGATGCGGGAGGA AGG
High GC content Enzymes: TauI, Fsp4HI
Cloning / PCR primers
|
86 | 96 | -- | -- | -- | -- |
0 - 0 - 0 - 3 - 61
0 - 0 - 0 - 1 - 0 64 off-targets |
show all... |
42 / fw |
TGCCGCCTTCCTCCCGCATC CGG
Enzymes: LweI, MwoI, MspI, HinP1I, LpnPI, BslI, BspFNI
Cloning / PCR primers
|
86 | 94 | -- | -- | -- | -- |
0 - 0 - 1 - 9 - 85
0 - 0 - 0 - 1 - 2 95 off-targets |
show all... |
90 / rev |
CCGGGTCGCGGGGACGGCGC GGG
High GC content Enzymes: HinP1I, BspFNI, MwoI, BceAI, BslFI
Cloning / PCR primers
|
86 | 92 | 28 | 78 | 74 | 70 |
0 - 0 - 0 - 17 - 93
0 - 0 - 0 - 2 - 2 110 off-targets |
show all... |
101 / rev |
GTGAGCGCGGCCCGGGTCGC GGG
High GC content Enzymes: BspFNI, MwoI, BslFI, BslI
Cloning / PCR primers
|
85 | 95 | -- | -- | -- | -- |
0 - 0 - 1 - 6 - 52
0 - 0 - 0 - 0 - 0 59 off-targets |
show all... |
72 / fw |
GCCATGGTGCGGTTCAAGCA CGG
Enzymes: MwoI
Cloning / PCR primers
|
84 | 93 | 55 | 46 | 66 | 88 |
0 - 0 - 2 - 7 - 64
0 - 0 - 0 - 0 - 2 73 off-targets |
show all... |
110 / fw |
CCCGCGCCGTCCCCGCGACC CGG
High GC content Enzymes: BshFI, BmeT110I, TauI, MwoI, MspI, BseDI, PspPI, NciI, LpnPI, BslI, StyD4I, SsiI, Cfr9I, Fsp4HI
Cloning / PCR primers
|
84 | 94 | -- | -- | -- | -- |
0 - 0 - 0 - 12 - 87
0 - 0 - 0 - 1 - 0 99 off-targets |
show all... |
12 / rev |
GAGGAAGGCGGCACTAGGCT CGG
Enzymes: Hpy188I
Cloning / PCR primers
|
81 | 89 | -- | -- | -- | -- |
0 - 0 - 0 - 12 - 113
0 - 0 - 0 - 0 - 0 125 off-targets |
show all... |
17 / rev |
TGCGGGAGGAAGGCGGCACT AGG
Enzymes: MaeI
Cloning / PCR primers
|
81 | 90 | -- | -- | -- | -- |
0 - 0 - 2 - 12 - 133
0 - 0 - 0 - 4 - 2 147 off-targets |
show all... |
114 / rev |
CGGCGACAGCGGGGTGAGCG CGG
High GC content Enzymes: BshFI, TauI, MwoI, NciI, MspI, PspPI, HinP1I, BspFNI, LpnPI, StyD4I, Fsp4HI
Cloning / PCR primers
|
81 | 93 | -- | -- | -- | -- |
0 - 0 - 0 - 17 - 88
0 - 0 - 0 - 1 - 1 105 off-targets |
show all... |
102 / rev |
GGTGAGCGCGGCCCGGGTCG CGG
High GC content Enzymes: BspFNI, MwoI, BslFI, BslI
Cloning / PCR primers
|
80 | 92 | -- | -- | -- | -- |
0 - 0 - 3 - 7 - 101
0 - 0 - 1 - 0 - 0 111 off-targets |
show all... |
91 / rev |
CCCGGGTCGCGGGGACGGCG CGG
High GC content Enzymes: HinP1I, BspFNI, MwoI, BceAI, BslFI
Cloning / PCR primers
|
79 | 86 | 26 | 62 | 78 | 77 |
0 - 0 - 1 - 12 - 102
0 - 0 - 0 - 2 - 5 115 off-targets |
show all... |
96 / rev |
CGCGGCCCGGGTCGCGGGGA CGG
High GC content Enzymes: HinP1I, BspFNI, MwoI, BceAI, BslFI
Cloning / PCR primers
|
79 | 89 | -- | -- | -- | -- |
0 - 0 - 0 - 8 - 119
0 - 0 - 0 - 1 - 1 127 off-targets |
show all... |
89 / rev |
CGGGTCGCGGGGACGGCGCG GGG
High GC content Enzymes: HinP1I, BspFNI, MwoI, BslFI
Cloning / PCR primers
|
78 | 93 | 30 | 78 | 74 | 87 |
0 - 0 - 2 - 9 - 109
0 - 0 - 0 - 2 - 2 120 off-targets |
show all... |
108 / rev |
CAGCGGGGTGAGCGCGGCCC GGG
High GC content Enzymes: BshFI, BmeT110I, TauI, MwoI, NciI, MspI, BseDI, PspPI, BspFNI, LpnPI, StyD4I, Cfr9I, Fsp4HI
Cloning / PCR primers
|
77 | 90 | -- | -- | -- | -- |
0 - 0 - 2 - 11 - 105
0 - 0 - 0 - 0 - 0 118 off-targets |
show all... |
111 / fw |
CCGCGCCGTCCCCGCGACCC GGG
High GC content Enzymes: BshFI, BmeT110I, TauI, MwoI, NciI, MspI, BseDI, PspPI, BspFNI, LpnPI, BslI, StyD4I, SsiI, Cfr9I, Fsp4HI
Cloning / PCR primers
|
76 | 94 | -- | -- | -- | -- |
0 - 0 - 2 - 25 - 85
0 - 0 - 0 - 16 - 0 112 off-targets |
show all... |
24 / rev |
CGCCGGATGCGGGAGGAAGG CGG
Enzymes: TauI, Fsp4HI
Cloning / PCR primers
|
66 | 86 | -- | -- | -- | -- |
0 - 0 - 0 - 8 - 114
0 - 0 - 0 - 2 - 8 122 off-targets |
show all... |
73 / fw |
CCATGGTGCGGTTCAAGCAC GGG
Enzymes: MwoI
Cloning / PCR primers
|
59 | 93 | 51 | 54 | 67 | 77 |
0 - 1 - 0 - 5 - 43
0 - 0 - 0 - 1 - 0 49 off-targets |
show all... |
53 / rev |
CCCGTGCTTGAACCGCACCA TGG
Enzymes: BstDSI, AcyI, BseDI, ErhI, NlaIII, BslI, Bsp19I
Cloning / PCR primers
|
49 | 97 | 70 | 54 | 74 | 76 |
0 - 1 - 1 - 1 - 20
0 - 1 - 0 - 0 - 1 23 off-targets |
show all... |