Job name: hilA gene against S. Typhimurium LT2

Candidate gRNAs

• On-target score is evaluated based on the physical and sequence features of gRNA.
• The score is ranged from 0 to 1. Higher score indicates higher predicted on-target activity.
• Off-target score use CFD (Cutting Frequency Determination) score generated by Doench et. al., (2016) to predict off-target activity of gRNAs.
• Click + for more details on the off-targets.
• The score is ranged from 0 to 1. Higher score indicates higher off-target activity
• *Best gRNA would have lower off-target score

gRNA sequence	Position	GC_20nt (%)	Tm_20nt (°C)	Model 3 Score	Model 4 score	Off-target
TGATGATTTCATACTCAACA TGG	3019909 - 3019929	30.0	43.95	1.00000	0.99971	2
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
	TGAAGGTTTCAAACTCGACC TGG		5	110324	110347	0.010913
	TAATGATTCCATGCTCAAAA TGG		4	4177590	4177613	0.218310
GATTTCATACTCAACATGGA CGG	3019913 - 3019933	35.0	44.80	0.99999	0.99738	2
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
	GGTTTCAAACTCGACCTGGG CGG		5	110328	110351	0.000000
	GATTTTATAATCAACATAGG TGG		5	1549757	1549780	0.426050
GCCAAAAGAATATGCCGTTC TGG	3019972 - 3019992	45.0	50.07	0.99810	0.64083	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
GTCATCCTGCTCGAAGCCGC CGG	3019994 - 3020014	65.0	58.15	0.99999	0.99746	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
TGTGAGTAAAAACACCTTAC TGG	3020023 - 3020043	35.0	45.26	0.99997	0.99221	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
TAAAAACACCTTACTGGACC AGG	3020029 - 3020049	40.0	46.94	0.97512	0.23554	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
ACACCTTACTGGACCAGGTA TGG	3020034 - 3020054	50.0	51.31	0.99847	0.28685	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
CACCTTACTGGACCAGGTAT GGG	3020035 - 3020055	50.0	50.04	0.99942	0.21908	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
ACCTTACTGGACCAGGTATG GGG	3020036 - 3020056	50.0	50.04	0.99989	0.48812	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
GGACCAGGTATGGGGCGACG CGG	3020044 - 3020064	70.0	58.93	0.23439	0.00733	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
TGCCTTACGACGTATTCTGT CGG	3020101 - 3020121	45.0	50.94	1.00000	0.99698	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
CGTTACATTGAAACACTGTA CGG	3020138 - 3020158	35.0	45.90	0.99999	0.99678	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
CATTGAAACACTGTACGGAC AGG	3020143 - 3020163	45.0	49.08	0.99962	0.97610	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
ATTGAAACACTGTACGGACA GGG	3020144 - 3020164	40.0	49.17	0.99994	0.99722	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
CACTGTACGGACAGGGCTAT CGG	3020151 - 3020171	55.0	53.06	0.98680	0.28352	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
GGGCTATCGGTTTAATCGTC CGG	3020164 - 3020184	50.0	50.46	0.99396	0.11109	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
GTTTAATCGTCCGGTCGTAG TGG	3020173 - 3020193	50.0	50.54	0.99922	0.65626	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
GCAACCTACGACTCATACAT TGG	3020212 - 3020232	45.0	49.28	0.99999	0.99865	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
GATACTTCCTTTTCAGATGC AGG	3020236 - 3020256	40.0	46.01	0.99998	0.91496	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
TCCTTTTCAGATGCAGGATC AGG	3020242 - 3020262	45.0	49.32	0.96385	0.04865	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
TCTGCATTACTCTATCGTGA AGG	3020278 - 3020298	40.0	47.58	1.00000	0.99948	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
CTGCATTACTCTATCGTGAA GGG	3020279 - 3020299	40.0	46.92	1.00000	0.99905	1
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
	CTGCATTTCCCGATCGGGAT GGG		5	2267175	2267198	0.008067
TTATCGCAGTATGCGCCCTT TGG	3020303 - 3020323	50.0	54.09	0.99877	0.35626	2
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
	TTATCGCAATATGCGCATTC AGG		5	277761	277784	0.017532
	TTTTCTCATTATGCGCCAGT TGG		5	624413	624436	0.033786
CTTTGGCCTGAGCGTGCTGC CGG	3020320 - 3020340	65.0	59.16	1.00000	0.99863	2
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
	CTTCGGTCTGATCGTGCTTC TGG		5	2562269	2562292	0.134804
	ATATGGCCTGGCCATGCTGC CGG		5	3198555	3198578	0.184349
GAAGAACTGCCGCAGTGTTA AGG	3020353 - 3020373	50.0	52.79	0.99979	0.93782	3
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
	GAGGAACTGGCGCAATGGTT AGG		5	861479	861502	0.044735
	CAAGAAATGCCGCCGTGGGA AGG		5	1456904	1456927	0.009566
	GAAGACCTGCCGCTGTTCAA AGG		5	4627975	4627998	0.007421
TAAGGATATTCTTGAGCTCA TGG	3020371 - 3020391	35.0	44.79	0.99998	0.99184	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
CGCCCCGATTATTATATCTC CGG	3020402 - 3020422	45.0	47.30	0.99994	0.88925	2
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
	GGCCCCGTTTTTTATTTCTC TGG		5	1931242	1931265	0.000000
	CGCGCCGATTTTTACACCTT CGG		5	4829982	4830005	0.008885
GCCCCGATTATTATATCTCC GGG	3020403 - 3020423	45.0	46.75	0.99931	0.43169	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
TCCGGGCAGATGATACCCGA TGG	3020420 - 3020440	60.0	56.68	0.85408	0.02102	2
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
	TCCGCATTGATGATACCCGC TGG		5	645831	645854	0.062181
	TGCGGGCAGGTGATGCACTA TGG		5	1753959	1753982	0.078154
TTGTACAGATTGAGATAGTT CGG	3020454 - 3020474	30.0	42.93	1.00000	0.99957	1
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
	ATGTACAGTTCGATATCGTT CGG		5	2711519	2711542	0.018824
TGTACAGATTGAGATAGTTC GGG	3020455 - 3020475	35.0	43.61	1.00000	0.99971	2
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
	TGTACAGTTCGATATCGTTC GGG		4	2711520	2711543	0.000000
	TGCATTGATTCAGACAGTTC GGG		5	4691632	4691655	0.022769
ATTGAGATAGTTCGGGTTAA AGG	3020462 - 3020482	35.0	45.46	0.99607	0.45529	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
AGGTTATCACCTGCTGCACC AGG	3020482 - 3020502	55.0	54.17	0.99991	0.51962	1
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
	CGGTTATCGTCTGGTGTACC AGG		5	4691870	4691893	0.000000
CTTTTGCTCAGATGTATTCC CGG	3020564 - 3020584	40.0	46.30	1.00000	0.99185	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
GATGTATTCCCGGACTTCGC TGG	3020574 - 3020594	55.0	52.83	0.99999	0.95798	1
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
	GATGTATTTCCGGATGGCGC CGG		5	993723	993746	0.002376
ATGTATTCCCGGACTTCGCT GGG	3020575 - 3020595	50.0	53.02	0.99999	0.97459	1
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
	AGGTATACCTGGAATGCGCT GGG		5	334600	334623	0.083810
AAATTCGATTGACAGTACTA TGG	3020620 - 3020640	30.0	43.78	1.00000	0.99995	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
AGTACTATGGTTTACTTACG CGG	3020633 - 3020653	35.0	43.68	1.00000	0.99613	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
CATTGCGCCTTACTGTGCGC TGG	3020743 - 3020763	60.0	57.57	0.99852	0.42265	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
GGCAGAATGCTACCTCAGCA TGG	3020764 - 3020784	55.0	54.28	0.98257	0.29086	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
CTACCTCAGCATGGCGCAAA TGG	3020773 - 3020793	55.0	55.25	0.99999	0.99784	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
TACCTCAGCATGGCGCAAAT GGG	3020774 - 3020794	50.0	54.54	0.99945	0.91680	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
ACCTCAGCATGGCGCAAATG GGG	3020775 - 3020795	55.0	56.16	0.99973	0.95952	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
AGCTAAAGAACATGCGATTA AGG	3020827 - 3020847	35.0	46.98	1.00000	0.99988	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
TGCGATTAAGGCGACAGAGC TGG	3020839 - 3020859	55.0	55.11	1.00000	0.99925	1
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
	GGCGTTAAAGGCCACAAAGC TGG		5	502265	502288	0.119603
CACAATAATCCACAAGCTTT AGG	3020864 - 3020884	35.0	46.30	0.99991	0.99218	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
TCCACAAGCTTTAGGATTAC TGG	3020872 - 3020892	40.0	46.83	0.99758	0.55684	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
CCACAAGCTTTAGGATTACT GGG	3020873 - 3020893	40.0	46.80	0.99959	0.84142	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
CACAAGCTTTAGGATTACTG GGG	3020874 - 3020894	40.0	45.95	0.99976	0.80360	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
ATTCACTCAGAATACATCGT CGG	3020909 - 3020929	35.0	45.97	1.00000	0.99989	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
TTCACTCAGAATACATCGTC GGG	3020910 - 3020930	40.0	46.86	1.00000	0.99856	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
GCAGATATTAAATATTATTA TGG	3020975 - 3020995	15.0	34.84	1.00000	0.96794	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
ATATTAAATATTATTATGGC TGG	3020979 - 3020999	15.0	35.04	1.00000	0.98759	4
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
	ATACTGAACATTATCATGAC TGG		5	1826407	1826430	0.083110
	ATATTAAAGACTATGACGAC TGG		5	3444284	3444307	0.010404
	TTACAAAATATGATTTTGGC TGG		5	3809012	3809035	0.000000
	ATATGAATTATAATTAAAGC TGG		5	4777010	4777033	0.081020
CATGGCTGGTCAGTTGGAGG AGG	3021010 - 3021030	60.0	54.85	0.99151	0.27434	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
GATTAACGAGTGTTTAAAAT TGG	3021043 - 3021063	25.0	41.67	1.00000	0.99545	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
CCGCAGGGATCACTAAGCTG TGG	3021081 - 3021101	60.0	54.78	0.99993	0.95377	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
TGGATTACCTATTATCATAC CGG	3021101 - 3021121	30.0	40.43	1.00000	0.97002	2
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
	GGCATTACCCAGTACCATAC CGG		5	2347709	2347732	0.026756
	GGCATTACCCAGTACCATAC CGG		5	2347709	2347732	0.026756
ATTGATGATGCTATACGTTT AGG	3021125 - 3021145	30.0	44.56	1.00000	0.99990	1
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
	CTTGATGAGGCTATGCGTAT TGG		5	2956626	2956649	0.098542
TCCAATATTATTAAGTATGC AGG	3021181 - 3021201	25.0	39.68	1.00000	0.99562	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
GTTATGTTTCTTTCGCTTAA AGG	3021203 - 3021223	30.0	44.13	0.99943	0.24274	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
GCTTAAAGGTAAACATGAAC TGG	3021217 - 3021237	35.0	44.76	0.99986	0.97953	1
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
	GCATGAAGGTCAACATCAAC GGG		5	1123217	1123240	0.048403
AACTAAAGAAATATCCACGC AGG	3021250 - 3021270	35.0	45.60	0.99995	0.99818	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
CCGTTAGTGCTGGTTGCCCA CGG	3021401 - 3021421	60.0	57.18	0.98487	0.04616	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
AAGCTATTGCCGAGAAAATG TGG	3021426 - 3021446	40.0	48.66	0.99999	0.99830	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
TTAAAAACGAAGACAATATT TGG	3021456 - 3021476	20.0	40.68	0.99999	0.99836	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
ACAATATTTGGTTCAAAAGA TGG	3021468 - 3021488	25.0	42.82	1.00000	0.99914	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
TTGGTTCAAAAGATGGAAAC AGG	3021475 - 3021495	35.0	46.41	0.99994	0.97036	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score
ATCCCCGCTTGATTAAATTA CGG	3021498 - 3021518	35.0	46.43	0.99994	0.96015	0
	OFF-TARGET PREDICTION:
	Off-target		Mismatches	Start	End	CFD Score