PRIMER_THERMODYNAMIC_OLIGO_ALIGNMENT=1
PRIMER_THERMODYNAMIC_PARAMETERS_PATH=primer3_config/
PRIMER_TM_FORMULA=1
PRIMER_SALT_CORRECTIONS=1
PRIMER_SALT_DIVALENT=1.5
SEQUENCE_ID=Check left only
SEQUENCE_PRIMER=gtcctcgaacagctcagtcc
PRIMER_TASK=check_primers
PRIMER_PICK_ANYWAY=1
PRIMER_DNTP_CONC=0.0
PRIMER_WARNING=PRIMER_SALT_DIVALENT > 0.0 but PRIMER_DNTP_CONC <= 0.0; use reasonable value for PRIMER_DNTP_CONC
PRIMER_LEFT_NUM_RETURNED=1
PRIMER_RIGHT_NUM_RETURNED=0
PRIMER_INTERNAL_NUM_RETURNED=0
PRIMER_PAIR_NUM_RETURNED=0
PRIMER_LEFT_0_PENALTY=1.013677
PRIMER_LEFT_0_SEQUENCE=gtcctcgaacagctcagtcc
PRIMER_LEFT_0=0,20
PRIMER_LEFT_0_TM=61.014
PRIMER_LEFT_0_GC_PERCENT=60.000
PRIMER_LEFT_0_SELF_ANY_TH=0.00
PRIMER_LEFT_0_SELF_END_TH=0.00
PRIMER_LEFT_0_HAIRPIN_TH=0.00
PRIMER_LEFT_0_END_STABILITY=3.8500
=
SEQUENCE_ID=Check internal only
SEQUENCE_INTERNAL_OLIGO=ccacggatgaggaagaagaa
PRIMER_TASK=check_primers
PRIMER_PICK_ANYWAY=1
PRIMER_WARNING=PRIMER_SALT_DIVALENT > 0.0 but PRIMER_DNTP_CONC <= 0.0; use reasonable value for PRIMER_DNTP_CONC
PRIMER_LEFT_NUM_RETURNED=0
PRIMER_RIGHT_NUM_RETURNED=0
PRIMER_INTERNAL_NUM_RETURNED=1
PRIMER_PAIR_NUM_RETURNED=0
PRIMER_INTERNAL_0_PENALTY=8.869010
PRIMER_INTERNAL_0_PROBLEMS= Temperature too low;
PRIMER_INTERNAL_0_SEQUENCE=ccacggatgaggaagaagaa
PRIMER_INTERNAL_0=90,20
PRIMER_INTERNAL_0_TM=51.131
PRIMER_INTERNAL_0_GC_PERCENT=50.000
PRIMER_INTERNAL_0_SELF_ANY_TH=0.00
PRIMER_INTERNAL_0_SELF_END_TH=0.00
PRIMER_INTERNAL_0_HAIRPIN_TH=0.00
=
SEQUENCE_ID=Check right only
SEQUENCE_PRIMER_REVCOMP=gcgtttttattgccgttgtt
PRIMER_TASK=check_primers
PRIMER_PICK_ANYWAY=1
PRIMER_WARNING=PRIMER_SALT_DIVALENT > 0.0 but PRIMER_DNTP_CONC <= 0.0; use reasonable value for PRIMER_DNTP_CONC
PRIMER_LEFT_NUM_RETURNED=0
PRIMER_RIGHT_NUM_RETURNED=1
PRIMER_INTERNAL_NUM_RETURNED=0
PRIMER_PAIR_NUM_RETURNED=0
PRIMER_RIGHT_0_PENALTY=2.682567
PRIMER_RIGHT_0_SEQUENCE=gcgtttttattgccgttgtt
PRIMER_RIGHT_0=199,20
PRIMER_RIGHT_0_TM=57.317
PRIMER_RIGHT_0_GC_PERCENT=40.000
PRIMER_RIGHT_0_SELF_ANY_TH=0.00
PRIMER_RIGHT_0_SELF_END_TH=0.00
PRIMER_RIGHT_0_HAIRPIN_TH=0.00
PRIMER_RIGHT_0_END_STABILITY=2.8300
=
SEQUENCE_ID=Pick the same primers as before
SEQUENCE_TEMPLATE=atggatttgatggacatggatgccatcaatgacctccacttctctggacacagctccctcaccaacacccctacttccgacgaggattatggttgcacctggaaccactggtctcccatcgtcaactgggacacctttacaggtgccccagatgactttcaccacctcatggacaccatcatcgaggacagaacaactgtcctcgaacagctcagtccatctatcaccacaaccaccaccaccaccacaaccacggatgaggaagaagaagaaatggaaacaacaaccacaacaacaacaacggcaataaaaacgcatgaagttggtgatgattcaaaagggctaaaactagtgcacctattgatggctggcgcggaagccttaaccggttcaaccaaaaaccgtgacttagctcgagtgatattgatccggctcaaggaattagtctcacaacatgctaacggctccaacatggagagacttgccgctcatttcaccgaagcccttcatggactactagaaggtgctgggggtgcgcataataaccaccatcaccacaacaacaacaaacattacctcaccacaaatggtccccacgacaaccaaaacgacacacttgctgctttccaactacttcaagacatgtcaccttacgtcaaattcggccacttcacagccaatcaagccatcatcgaagccgtggcccacgaacgccgcgtccatgtcatcgactacgatatcatggaaggggtccaatgggcctcattaatccaatcacttgcttccaacaacaacggtccacatcttcggataaccgcattatcgcggacaggaaccggccggaggtcaatcgccaccgtacaagaaaccggcaggcgattaacctcctttgccgcttccctcggacaaccattttcttttcatcactgcagattagactccgatgaaactttccgtccttctgcattaaagcttgtacgtggagaggctttggttttcaactgcatgctgaatttacctcaccttagttaccgtgcaccggaatcagttgcttcgtttttaaacggagcaaaaacgttgaatccaaagcttgtgacgttggttgaagaagaagttggttctgttattggtgggtttgtggaaaggttcatggactcacttcatcattattcagcggtttttgattcattggaggctggttttccgatgcagaaccgggcccggactttggtggagagggtattttttgggcctagaattgcaggctcgttgggccggatatatagaacgggtggtgaagaggagagaaggtcatggggggagtggttaggtgaggtagggtttaggggagttccggtgagctttgcaaatcattgtcaagcaaagctgttgttagggctttttaatgatgggtatagggttgaagaggtgggtgtgggtagtaataagttggtgttggattggaaatcaagacgtttgctttctgcttccctttggacttgttcttcttcagattctgatttatag
PRIMER_LOWERCASE_MASKING=0
PRIMER_TASK=pick_detection_primers
PRIMER_PICK_RIGHT_PRIMER=1
PRIMER_PICK_INTERNAL_OLIGO=1
PRIMER_PICK_LEFT_PRIMER=1
PRIMER_NUM_RETURN=1
PRIMER_WARNING=PRIMER_SALT_DIVALENT > 0.0 but PRIMER_DNTP_CONC <= 0.0; use reasonable value for PRIMER_DNTP_CONC
PRIMER_LEFT_NUM_RETURNED=1
PRIMER_RIGHT_NUM_RETURNED=1
PRIMER_INTERNAL_NUM_RETURNED=1
PRIMER_PAIR_NUM_RETURNED=1
PRIMER_PAIR_0_PENALTY=0.055158
PRIMER_LEFT_0_PENALTY=0.029777
PRIMER_RIGHT_0_PENALTY=0.025380
PRIMER_INTERNAL_0_PENALTY=0.088782
PRIMER_LEFT_0_SEQUENCE=acgtcaaattcggccacttc
PRIMER_RIGHT_0_SEQUENCE=atccgaagatgtggaccgtt
PRIMER_INTERNAL_0_SEQUENCE=cgccgcgtccatgtcatcga
PRIMER_LEFT_0=652,20
PRIMER_RIGHT_0=811,20
PRIMER_INTERNAL_0=711,20
PRIMER_LEFT_0_TM=60.030
PRIMER_RIGHT_0_TM=60.025
PRIMER_INTERNAL_0_TM=59.911
PRIMER_LEFT_0_GC_PERCENT=50.000
PRIMER_RIGHT_0_GC_PERCENT=50.000
PRIMER_INTERNAL_0_GC_PERCENT=65.000
PRIMER_INTERNAL_0_SELF_ANY_TH=2.84
PRIMER_LEFT_0_SELF_ANY_TH=2.58
PRIMER_RIGHT_0_SELF_ANY_TH=0.00
PRIMER_INTERNAL_0_SELF_END_TH=0.00
PRIMER_LEFT_0_SELF_END_TH=0.00
PRIMER_RIGHT_0_SELF_END_TH=0.00
PRIMER_LEFT_0_HAIRPIN_TH=0.00
PRIMER_RIGHT_0_HAIRPIN_TH=37.69
PRIMER_INTERNAL_0_HAIRPIN_TH=0.00
PRIMER_LEFT_0_END_STABILITY=3.0100
PRIMER_RIGHT_0_END_STABILITY=4.4400
PRIMER_PAIR_0_COMPL_ANY_TH=17.70
PRIMER_PAIR_0_COMPL_END_TH=17.70
PRIMER_PAIR_0_PRODUCT_SIZE=160
PRIMER_PAIR_0_PRODUCT_TM=87.3
=
SEQUENCE_ID=Check primers with sequence
SEQUENCE_TEMPLATE=atggatttgatggacatggatgccatcaatgacctccacttctctggacacagctccctcaccaacacccctacttccgacgaggattatggttgcacctggaaccactggtctcccatcgtcaactgggacacctttacaggtgccccagatgactttcaccacctcatggacaccatcatcgaggacagaacaactgtcctcgaacagctcagtccatctatcaccacaaccaccaccaccaccacaaccacggatgaggaagaagaagaaatggaaacaacaaccacaacaacaacaacggcaataaaaacgcatgaagttggtgatgattcaaaagggctaaaactagtgcacctattgatggctggcgcggaagccttaaccggttcaaccaaaaaccgtgacttagctcgagtgatattgatccggctcaaggaattagtctcacaacatgctaacggctccaacatggagagacttgccgctcatttcaccgaagcccttcatggactactagaaggtgctgggggtgcgcataataaccaccatcaccacaacaacaacaaacattacctcaccacaaatggtccccacgacaaccaaaacgacacacttgctgctttccaactacttcaagacatgtcaccttacgtcaaattcggccacttcacagccaatcaagccatcatcgaagccgtggcccacgaacgccgcgtccatgtcatcgactacgatatcatggaaggggtccaatgggcctcattaatccaatcacttgcttccaacaacaacggtccacatcttcggataaccgcattatcgcggacaggaaccggccggaggtcaatcgccaccgtacaagaaaccggcaggcgattaacctcctttgccgcttccctcggacaaccattttcttttcatcactgcagattagactccgatgaaactttccgtccttctgcattaaagcttgtacgtggagaggctttggttttcaactgcatgctgaatttacctcaccttagttaccgtgcaccggaatcagttgcttcgtttttaaacggagcaaaaacgttgaatccaaagcttgtgacgttggttgaagaagaagttggttctgttattggtgggtttgtggaaaggttcatggactcacttcatcattattcagcggtttttgattcattggaggctggttttccgatgcagaaccgggcccggactttggtggagagggtattttttgggcctagaattgcaggctcgttgggccggatatatagaacgggtggtgaagaggagagaaggtcatggggggagtggttaggtgaggtagggtttaggggagttccggtgagctttgcaaatcattgtcaagcaaagctgttgttagggctttttaatgatgggtatagggttgaagaggtgggtgtgggtagtaataagttggtgttggattggaaatcaagacgtttgctttctgcttccctttggacttgttcttcttcagattctgatttatag
PRIMER_PICK_RIGHT_PRIMER=0
PRIMER_PICK_INTERNAL_OLIGO=0
PRIMER_PICK_LEFT_PRIMER=0
PRIMER_NUM_RETURN=5
PRIMER_TASK=check_primers
SEQUENCE_PRIMER=gtcctcgaacagctcagtcc
SEQUENCE_INTERNAL_OLIGO=ccacggatgaggaagaagaa
SEQUENCE_PRIMER_REVCOMP=gcgtttttattgccgttgtt
PRIMER_PICK_ANYWAY=1
PRIMER_WARNING=PRIMER_SALT_DIVALENT > 0.0 but PRIMER_DNTP_CONC <= 0.0; use reasonable value for PRIMER_DNTP_CONC
PRIMER_LEFT_NUM_RETURNED=1
PRIMER_RIGHT_NUM_RETURNED=1
PRIMER_INTERNAL_NUM_RETURNED=1
PRIMER_PAIR_NUM_RETURNED=1
PRIMER_PAIR_0_PENALTY=3.696244
PRIMER_LEFT_0_PENALTY=1.013677
PRIMER_RIGHT_0_PENALTY=2.682567
PRIMER_INTERNAL_0_PENALTY=8.869010
PRIMER_INTERNAL_0_PROBLEMS= Temperature too low;
PRIMER_LEFT_0_SEQUENCE=gtcctcgaacagctcagtcc
PRIMER_RIGHT_0_SEQUENCE=gcgtttttattgccgttgtt
PRIMER_INTERNAL_0_SEQUENCE=ccacggatgaggaagaagaa
PRIMER_LEFT_0=198,20
PRIMER_RIGHT_0=315,20
PRIMER_INTERNAL_0=250,20
PRIMER_LEFT_0_TM=61.014
PRIMER_RIGHT_0_TM=57.317
PRIMER_INTERNAL_0_TM=51.131
PRIMER_LEFT_0_GC_PERCENT=60.000
PRIMER_RIGHT_0_GC_PERCENT=40.000
PRIMER_INTERNAL_0_GC_PERCENT=50.000
PRIMER_INTERNAL_0_SELF_ANY_TH=0.00
PRIMER_LEFT_0_SELF_ANY_TH=0.00
PRIMER_RIGHT_0_SELF_ANY_TH=0.00
PRIMER_INTERNAL_0_SELF_END_TH=0.00
PRIMER_LEFT_0_SELF_END_TH=0.00
PRIMER_RIGHT_0_SELF_END_TH=0.00
PRIMER_LEFT_0_HAIRPIN_TH=0.00
PRIMER_RIGHT_0_HAIRPIN_TH=0.00
PRIMER_INTERNAL_0_HAIRPIN_TH=0.00
PRIMER_LEFT_0_END_STABILITY=3.8500
PRIMER_RIGHT_0_END_STABILITY=2.8300
PRIMER_PAIR_0_COMPL_ANY_TH=0.00
PRIMER_PAIR_0_COMPL_END_TH=0.00
PRIMER_PAIR_0_PRODUCT_SIZE=118
PRIMER_PAIR_0_PRODUCT_TM=84.2
=
SEQUENCE_ID=Check primers without sequence
PRIMER_PICK_RIGHT_PRIMER=0
PRIMER_PICK_INTERNAL_OLIGO=0
PRIMER_PICK_LEFT_PRIMER=0
PRIMER_NUM_RETURN=5
PRIMER_TASK=check_primers
SEQUENCE_PRIMER=gtcctcgaacagctcagtcc
SEQUENCE_INTERNAL_OLIGO=ccacggatgaggaagaagaa
SEQUENCE_PRIMER_REVCOMP=gcgtttttattgccgttgtt
PRIMER_PICK_ANYWAY=1
PRIMER_WARNING=PRIMER_SALT_DIVALENT > 0.0 but PRIMER_DNTP_CONC <= 0.0; use reasonable value for PRIMER_DNTP_CONC
PRIMER_LEFT_NUM_RETURNED=1
PRIMER_RIGHT_NUM_RETURNED=1
PRIMER_INTERNAL_NUM_RETURNED=1
PRIMER_PAIR_NUM_RETURNED=1
PRIMER_PAIR_0_PENALTY=3.696244
PRIMER_LEFT_0_PENALTY=1.013677
PRIMER_RIGHT_0_PENALTY=2.682567
PRIMER_INTERNAL_0_PENALTY=8.869010
PRIMER_INTERNAL_0_PROBLEMS= Temperature too low;
PRIMER_LEFT_0_SEQUENCE=gtcctcgaacagctcagtcc
PRIMER_RIGHT_0_SEQUENCE=gcgtttttattgccgttgtt
PRIMER_INTERNAL_0_SEQUENCE=ccacggatgaggaagaagaa
PRIMER_LEFT_0=0,20
PRIMER_RIGHT_0=199,20
PRIMER_INTERNAL_0=90,20
PRIMER_LEFT_0_TM=61.014
PRIMER_RIGHT_0_TM=57.317
PRIMER_INTERNAL_0_TM=51.131
PRIMER_LEFT_0_GC_PERCENT=60.000
PRIMER_RIGHT_0_GC_PERCENT=40.000
PRIMER_INTERNAL_0_GC_PERCENT=50.000
PRIMER_INTERNAL_0_SELF_ANY_TH=0.00
PRIMER_LEFT_0_SELF_ANY_TH=0.00
PRIMER_RIGHT_0_SELF_ANY_TH=0.00
PRIMER_INTERNAL_0_SELF_END_TH=0.00
PRIMER_LEFT_0_SELF_END_TH=0.00
PRIMER_RIGHT_0_SELF_END_TH=0.00
PRIMER_LEFT_0_HAIRPIN_TH=0.00
PRIMER_RIGHT_0_HAIRPIN_TH=0.00
PRIMER_INTERNAL_0_HAIRPIN_TH=0.00
PRIMER_LEFT_0_END_STABILITY=3.8500
PRIMER_RIGHT_0_END_STABILITY=2.8300
PRIMER_PAIR_0_COMPL_ANY_TH=0.00
PRIMER_PAIR_0_COMPL_END_TH=0.00
PRIMER_PAIR_0_PRODUCT_SIZE=200
PRIMER_PAIR_0_PRODUCT_TM=72.9
=
SEQUENCE_ID=Check primers without sequence and internal
PRIMER_PICK_RIGHT_PRIMER=0
PRIMER_PICK_INTERNAL_OLIGO=0
PRIMER_PICK_LEFT_PRIMER=0
PRIMER_NUM_RETURN=5
PRIMER_TASK=check_primers
SEQUENCE_PRIMER=gtcctcgaacagctcagtcc
SEQUENCE_PRIMER_REVCOMP=gcgtttttattgccgttgtt
PRIMER_PICK_ANYWAY=1
PRIMER_WARNING=PRIMER_SALT_DIVALENT > 0.0 but PRIMER_DNTP_CONC <= 0.0; use reasonable value for PRIMER_DNTP_CONC
PRIMER_LEFT_NUM_RETURNED=1
PRIMER_RIGHT_NUM_RETURNED=1
PRIMER_INTERNAL_NUM_RETURNED=0
PRIMER_PAIR_NUM_RETURNED=1
PRIMER_PAIR_0_PENALTY=3.696244
PRIMER_LEFT_0_PENALTY=1.013677
PRIMER_RIGHT_0_PENALTY=2.682567
PRIMER_LEFT_0_SEQUENCE=gtcctcgaacagctcagtcc
PRIMER_RIGHT_0_SEQUENCE=gcgtttttattgccgttgtt
PRIMER_LEFT_0=0,20
PRIMER_RIGHT_0=199,20
PRIMER_LEFT_0_TM=61.014
PRIMER_RIGHT_0_TM=57.317
PRIMER_LEFT_0_GC_PERCENT=60.000
PRIMER_RIGHT_0_GC_PERCENT=40.000
PRIMER_LEFT_0_SELF_ANY_TH=0.00
PRIMER_RIGHT_0_SELF_ANY_TH=0.00
PRIMER_LEFT_0_SELF_END_TH=0.00
PRIMER_RIGHT_0_SELF_END_TH=0.00
PRIMER_LEFT_0_HAIRPIN_TH=0.00
PRIMER_RIGHT_0_HAIRPIN_TH=0.00
PRIMER_LEFT_0_END_STABILITY=3.8500
PRIMER_RIGHT_0_END_STABILITY=2.8300
PRIMER_PAIR_0_COMPL_ANY_TH=0.00
PRIMER_PAIR_0_COMPL_END_TH=0.00
PRIMER_PAIR_0_PRODUCT_SIZE=200
PRIMER_PAIR_0_PRODUCT_TM=70.9
=
SEQUENCE_ID=Pick three times the same primers
SEQUENCE_TEMPLATE=atggatttgatggacatggatgccatcaatgacctccacttctctggacacagctccctcaccaacacccctacttccgacgaggattatggttgcacctggaaccactggtctcccatcgtcaactgggacacctttacaggtgccccagatgactttcaccacctcatggacaccatcatcgaggacagaacaactgtcctcgaacagctcagtccatctatcaccacaaccaccaccaccaccacaaccacggatgaggaagaagaagaaatggaaacaacaaccacaacaacaacaacggcaataaaaacgcatgaagttggtgatgattcaaaagggctaaaactagtgcacctattgatggctggcgcggaagccttaaccggttcaaccaaaaaccgtgacttagctcgagtgatattgatccggctcaaggaattagtctcacaacatgctaacggctccaacatggagagacttgccgctcatttcaccgaagcccttcatggactactagaaggtgctgggggtgcgcataataaccaccatcaccacaacaacaacaaacattacctcaccacaaatggtccccacgacaaccaaaacgacacacttgctgctttccaactacttcaagacatgtcaccttacgtcaaattcggccacttcacagccaatcaagccatcatcgaagccgtggcccacgaacgccgcgtccatgtcatcgactacgatatcatggaaggggtccaatgggcctcattaatccaatcacttgcttccaacaacaacggtccacatcttcggataaccgcattatcgcggacaggaaccggccggaggtcaatcgccaccgtacaagaaaccggcaggcgattaacctcctttgccgcttccctcggacaaccattttcttttcatcactgcagattagactccgatgaaactttccgtccttctgcattaaagcttgtacgtggagaggctttggttttcaactgcatgctgaatttacctcaccttagttaccgtgcaccggaatcagttgcttcgtttttaaacggagcaaaaacgttgaatccaaagcttgtgacgttggttgaagaagaagttggttctgttattggtgggtttgtggaaaggttcatggactcacttcatcattattcagcggtttttgattcattggaggctggttttccgatgcagaaccgggcccggactttggtggagagggtattttttgggcctagaattgcaggctcgttgggccggatatatagaacgggtggtgaagaggagagaaggtcatggggggagtggttaggtgaggtagggtttaggggagttccggtgagctttgcaaatcattgtcaagcaaagctgttgttagggctttttaatgatgggtatagggttgaagaggtgggtgtgggtagtaataagttggtgttggattggaaatcaagacgtttgctttctgcttccctttggacttgttcttcttcagattctgatttatag
PRIMER_LOWERCASE_MASKING=0
PRIMER_TASK=pick_primer_list
PRIMER_PICK_RIGHT_PRIMER=1
PRIMER_PICK_INTERNAL_OLIGO=1
PRIMER_PICK_LEFT_PRIMER=1
PRIMER_NUM_RETURN=5
PRIMER_WARNING=PRIMER_SALT_DIVALENT > 0.0 but PRIMER_DNTP_CONC <= 0.0; use reasonable value for PRIMER_DNTP_CONC
PRIMER_LEFT_NUM_RETURNED=5
PRIMER_RIGHT_NUM_RETURNED=5
PRIMER_INTERNAL_NUM_RETURNED=5
PRIMER_PAIR_NUM_RETURNED=0
PRIMER_LEFT_0_PENALTY=0.025380
PRIMER_RIGHT_0_PENALTY=0.025380
PRIMER_INTERNAL_0_PENALTY=0.042931
PRIMER_LEFT_0_SEQUENCE=aacggtccacatcttcggat
PRIMER_RIGHT_0_SEQUENCE=atccgaagatgtggaccgtt
PRIMER_INTERNAL_0_SEQUENCE=gccgcttccctcggacaacc
PRIMER_LEFT_0=792,20
PRIMER_RIGHT_0=811,20
PRIMER_INTERNAL_0=891,20
PRIMER_LEFT_0_TM=60.025
PRIMER_RIGHT_0_TM=60.025
PRIMER_INTERNAL_0_TM=60.043
PRIMER_LEFT_0_GC_PERCENT=50.000
PRIMER_RIGHT_0_GC_PERCENT=50.000
PRIMER_INTERNAL_0_GC_PERCENT=70.000
PRIMER_INTERNAL_0_SELF_ANY_TH=2.79
PRIMER_LEFT_0_SELF_ANY_TH=0.00
PRIMER_RIGHT_0_SELF_ANY_TH=0.00
PRIMER_INTERNAL_0_SELF_END_TH=0.00
PRIMER_LEFT_0_SELF_END_TH=0.00
PRIMER_RIGHT_0_SELF_END_TH=0.00
PRIMER_LEFT_0_HAIRPIN_TH=33.46
PRIMER_RIGHT_0_HAIRPIN_TH=37.69
PRIMER_INTERNAL_0_HAIRPIN_TH=45.87
PRIMER_LEFT_0_END_STABILITY=4.1800
PRIMER_RIGHT_0_END_STABILITY=4.4400
PRIMER_LEFT_1_PENALTY=0.025995
PRIMER_RIGHT_1_PENALTY=0.028504
PRIMER_INTERNAL_1_PENALTY=0.088782
PRIMER_LEFT_1_SEQUENCE=ctggtctcccatcgtcaact
PRIMER_RIGHT_1_SEQUENCE=gatttgcaaagctcaccgga
PRIMER_INTERNAL_1_SEQUENCE=cgccgcgtccatgtcatcga
PRIMER_LEFT_1=107,20
PRIMER_RIGHT_1=1371,20
PRIMER_INTERNAL_1=711,20
PRIMER_LEFT_1_TM=60.026
PRIMER_RIGHT_1_TM=60.029
PRIMER_INTERNAL_1_TM=59.911
PRIMER_LEFT_1_GC_PERCENT=55.000
PRIMER_RIGHT_1_GC_PERCENT=50.000
PRIMER_INTERNAL_1_GC_PERCENT=65.000
PRIMER_INTERNAL_1_SELF_ANY_TH=2.84
PRIMER_LEFT_1_SELF_ANY_TH=0.00
PRIMER_RIGHT_1_SELF_ANY_TH=20.53
PRIMER_INTERNAL_1_SELF_END_TH=0.00
PRIMER_LEFT_1_SELF_END_TH=0.00
PRIMER_RIGHT_1_SELF_END_TH=0.00
PRIMER_LEFT_1_HAIRPIN_TH=41.58
PRIMER_RIGHT_1_HAIRPIN_TH=0.00
PRIMER_INTERNAL_1_HAIRPIN_TH=0.00
PRIMER_LEFT_1_END_STABILITY=3.1600
PRIMER_RIGHT_1_END_STABILITY=5.1400
PRIMER_LEFT_2_PENALTY=0.028504
PRIMER_RIGHT_2_PENALTY=0.028504
PRIMER_INTERNAL_2_PENALTY=0.180197
PRIMER_LEFT_2_SEQUENCE=tccggtgagctttgcaaatc
PRIMER_RIGHT_2_SEQUENCE=ttctgcatcggaaaaccagc
PRIMER_INTERNAL_2_SEQUENCE=acgccgcgtccatgtcatcg
PRIMER_LEFT_2=1352,20
PRIMER_RIGHT_2=1213,20
PRIMER_INTERNAL_2=710,20
PRIMER_LEFT_2_TM=60.029
PRIMER_RIGHT_2_TM=60.029
PRIMER_INTERNAL_2_TM=60.180
PRIMER_LEFT_2_GC_PERCENT=50.000
PRIMER_RIGHT_2_GC_PERCENT=50.000
PRIMER_INTERNAL_2_GC_PERCENT=65.000
PRIMER_INTERNAL_2_SELF_ANY_TH=9.42
PRIMER_LEFT_2_SELF_ANY_TH=13.90
PRIMER_RIGHT_2_SELF_ANY_TH=0.00
PRIMER_INTERNAL_2_SELF_END_TH=0.00
PRIMER_LEFT_2_SELF_END_TH=9.52
PRIMER_RIGHT_2_SELF_END_TH=0.00
PRIMER_LEFT_2_HAIRPIN_TH=0.00
PRIMER_RIGHT_2_HAIRPIN_TH=0.00
PRIMER_INTERNAL_2_HAIRPIN_TH=27.20
PRIMER_LEFT_2_END_STABILITY=2.1700
PRIMER_RIGHT_2_END_STABILITY=4.8500
PRIMER_LEFT_3_PENALTY=0.028504
PRIMER_RIGHT_3_PENALTY=0.029777
PRIMER_INTERNAL_3_PENALTY=0.309918
PRIMER_LEFT_3_SEQUENCE=gctggttttccgatgcagaa
PRIMER_RIGHT_3_SEQUENCE=gaagtggccgaatttgacgt
PRIMER_INTERNAL_3_SEQUENCE=ggtgctgggggtgcgcataa
PRIMER_LEFT_3=1194,20
PRIMER_RIGHT_3=671,20
PRIMER_INTERNAL_3=522,20
PRIMER_LEFT_3_TM=60.029
PRIMER_RIGHT_3_TM=60.030
PRIMER_INTERNAL_3_TM=59.690
PRIMER_LEFT_3_GC_PERCENT=50.000
PRIMER_RIGHT_3_GC_PERCENT=50.000
PRIMER_INTERNAL_3_GC_PERCENT=65.000
PRIMER_INTERNAL_3_SELF_ANY_TH=12.28
PRIMER_LEFT_3_SELF_ANY_TH=0.00
PRIMER_RIGHT_3_SELF_ANY_TH=0.00
PRIMER_INTERNAL_3_SELF_END_TH=0.00
PRIMER_LEFT_3_SELF_END_TH=0.00
PRIMER_RIGHT_3_SELF_END_TH=0.00
PRIMER_LEFT_3_HAIRPIN_TH=44.71
PRIMER_RIGHT_3_HAIRPIN_TH=0.00
PRIMER_INTERNAL_3_HAIRPIN_TH=39.06
PRIMER_LEFT_3_END_STABILITY=3.0200
PRIMER_RIGHT_3_END_STABILITY=4.3400
PRIMER_LEFT_4_PENALTY=0.029777
PRIMER_RIGHT_4_PENALTY=0.029874
PRIMER_INTERNAL_4_PENALTY=0.381130
PRIMER_LEFT_4_SEQUENCE=acgtcaaattcggccacttc
PRIMER_RIGHT_4_SEQUENCE=tgaagtggccgaatttgacg
PRIMER_INTERNAL_4_SEQUENCE=cgaacgccgcgtccatgtca
PRIMER_LEFT_4=652,20
PRIMER_RIGHT_4=672,20
PRIMER_INTERNAL_4=707,20
PRIMER_LEFT_4_TM=60.030
PRIMER_RIGHT_4_TM=60.030
PRIMER_INTERNAL_4_TM=60.381
PRIMER_LEFT_4_GC_PERCENT=50.000
PRIMER_RIGHT_4_GC_PERCENT=50.000
PRIMER_INTERNAL_4_GC_PERCENT=65.000
PRIMER_INTERNAL_4_SELF_ANY_TH=16.23
PRIMER_LEFT_4_SELF_ANY_TH=2.58
PRIMER_RIGHT_4_SELF_ANY_TH=0.00
PRIMER_INTERNAL_4_SELF_END_TH=0.00
PRIMER_LEFT_4_SELF_END_TH=0.00
PRIMER_RIGHT_4_SELF_END_TH=0.00
PRIMER_LEFT_4_HAIRPIN_TH=0.00
PRIMER_RIGHT_4_HAIRPIN_TH=0.00
PRIMER_INTERNAL_4_HAIRPIN_TH=34.91
PRIMER_LEFT_4_END_STABILITY=3.0100
PRIMER_RIGHT_4_END_STABILITY=4.3500
=
SEQUENCE_ID=Make a list of a small range
SEQUENCE_TEMPLATE=atggatttgatggacatggatgccatcaatga
PRIMER_LOWERCASE_MASKING=0
PRIMER_TASK=pick_primer_list
PRIMER_PICK_RIGHT_PRIMER=1
PRIMER_PICK_INTERNAL_OLIGO=1
PRIMER_PICK_LEFT_PRIMER=1
PRIMER_NUM_RETURN=20
PRIMER_WARNING=PRIMER_SALT_DIVALENT > 0.0 but PRIMER_DNTP_CONC <= 0.0; use reasonable value for PRIMER_DNTP_CONC
PRIMER_LEFT_NUM_RETURNED=20
PRIMER_RIGHT_NUM_RETURNED=20
PRIMER_INTERNAL_NUM_RETURNED=4
PRIMER_PAIR_NUM_RETURNED=0
PRIMER_LEFT_0_PENALTY=1.380503
PRIMER_RIGHT_0_PENALTY=1.354059
PRIMER_INTERNAL_0_PENALTY=8.328764
PRIMER_LEFT_0_SEQUENCE=tggacatggatgccatcaatg
PRIMER_RIGHT_0_SEQUENCE=tgatggcatccatgtccatc
PRIMER_INTERNAL_0_SEQUENCE=tggatttgatggacatggatgccatca
PRIMER_LEFT_0=10,21
PRIMER_RIGHT_0=27,20
PRIMER_INTERNAL_0=1,27
PRIMER_LEFT_0_TM=59.619
PRIMER_RIGHT_0_TM=58.646
PRIMER_INTERNAL_0_TM=58.671
PRIMER_LEFT_0_GC_PERCENT=47.619
PRIMER_RIGHT_0_GC_PERCENT=50.000
PRIMER_INTERNAL_0_GC_PERCENT=44.444
PRIMER_INTERNAL_0_SELF_ANY_TH=20.68
PRIMER_LEFT_0_SELF_ANY_TH=19.55
PRIMER_RIGHT_0_SELF_ANY_TH=24.48
PRIMER_INTERNAL_0_SELF_END_TH=20.68
PRIMER_LEFT_0_SELF_END_TH=19.55
PRIMER_RIGHT_0_SELF_END_TH=23.68
PRIMER_LEFT_0_HAIRPIN_TH=44.12
PRIMER_RIGHT_0_HAIRPIN_TH=46.18
PRIMER_INTERNAL_0_HAIRPIN_TH=46.56
PRIMER_LEFT_0_END_STABILITY=2.8200
PRIMER_RIGHT_0_END_STABILITY=3.5100
PRIMER_LEFT_1_PENALTY=1.591479
PRIMER_RIGHT_1_PENALTY=1.354059
PRIMER_INTERNAL_1_PENALTY=8.471374
PRIMER_LEFT_1_SEQUENCE=tttgatggacatggatgcca
PRIMER_RIGHT_1_SEQUENCE=gatggcatccatgtccatca
PRIMER_INTERNAL_1_SEQUENCE=ggatttgatggacatggatgccatca
PRIMER_LEFT_1=5,20
PRIMER_RIGHT_1=26,20
PRIMER_INTERNAL_1=2,26
PRIMER_LEFT_1_TM=58.409
PRIMER_RIGHT_1_TM=58.646
PRIMER_INTERNAL_1_TM=57.529
PRIMER_LEFT_1_GC_PERCENT=45.000
PRIMER_RIGHT_1_GC_PERCENT=50.000
PRIMER_INTERNAL_1_GC_PERCENT=46.154
PRIMER_INTERNAL_1_SELF_ANY_TH=20.68
PRIMER_LEFT_1_SELF_ANY_TH=0.00
PRIMER_RIGHT_1_SELF_ANY_TH=24.61
PRIMER_INTERNAL_1_SELF_END_TH=20.68
PRIMER_LEFT_1_SELF_END_TH=0.00
PRIMER_RIGHT_1_SELF_END_TH=4.66
PRIMER_LEFT_1_HAIRPIN_TH=38.48
PRIMER_RIGHT_1_HAIRPIN_TH=46.18
PRIMER_INTERNAL_1_HAIRPIN_TH=46.56
PRIMER_LEFT_1_END_STABILITY=4.9200
PRIMER_RIGHT_1_END_STABILITY=3.0700
PRIMER_LEFT_2_PENALTY=1.657646
PRIMER_RIGHT_2_PENALTY=1.380503
PRIMER_INTERNAL_2_PENALTY=8.471374
PRIMER_LEFT_2_SEQUENCE=ggacatggatgccatcaatga
PRIMER_RIGHT_2_SEQUENCE=cattgatggcatccatgtcca
PRIMER_INTERNAL_2_SEQUENCE=tggatttgatggacatggatgccatc
PRIMER_LEFT_2=11,21
PRIMER_RIGHT_2=30,21
PRIMER_INTERNAL_2=1,26
PRIMER_LEFT_2_TM=59.342
PRIMER_RIGHT_2_TM=59.619
PRIMER_INTERNAL_2_TM=57.529
PRIMER_LEFT_2_GC_PERCENT=47.619
PRIMER_RIGHT_2_GC_PERCENT=47.619
PRIMER_INTERNAL_2_GC_PERCENT=46.154
PRIMER_INTERNAL_2_SELF_ANY_TH=18.44
PRIMER_LEFT_2_SELF_ANY_TH=26.38
PRIMER_RIGHT_2_SELF_ANY_TH=24.48
PRIMER_INTERNAL_2_SELF_END_TH=18.44
PRIMER_LEFT_2_SELF_END_TH=0.00
PRIMER_RIGHT_2_SELF_END_TH=2.36
PRIMER_LEFT_2_HAIRPIN_TH=44.12
PRIMER_RIGHT_2_HAIRPIN_TH=46.18
PRIMER_INTERNAL_2_HAIRPIN_TH=44.20
PRIMER_LEFT_2_END_STABILITY=2.5700
PRIMER_RIGHT_2_END_STABILITY=4.0200
PRIMER_LEFT_3_PENALTY=1.819575
PRIMER_RIGHT_3_PENALTY=1.591479
PRIMER_INTERNAL_3_PENALTY=9.328375
PRIMER_LEFT_3_SEQUENCE=tggacatggatgccatcaat
PRIMER_RIGHT_3_SEQUENCE=tggcatccatgtccatcaaa
PRIMER_INTERNAL_3_SEQUENCE=atggatttgatggacatggatgccatc
PRIMER_LEFT_3=10,20
PRIMER_RIGHT_3=24,20
PRIMER_INTERNAL_3=0,27
PRIMER_LEFT_3_TM=58.180
PRIMER_RIGHT_3_TM=58.409
PRIMER_INTERNAL_3_TM=57.672
PRIMER_LEFT_3_GC_PERCENT=45.000
PRIMER_RIGHT_3_GC_PERCENT=45.000
PRIMER_INTERNAL_3_GC_PERCENT=44.444
PRIMER_INTERNAL_3_SELF_ANY_TH=18.44
PRIMER_LEFT_3_SELF_ANY_TH=7.69
PRIMER_RIGHT_3_SELF_ANY_TH=0.00
PRIMER_INTERNAL_3_SELF_END_TH=18.44
PRIMER_LEFT_3_SELF_END_TH=0.00
PRIMER_RIGHT_3_SELF_END_TH=0.00
PRIMER_LEFT_3_HAIRPIN_TH=44.12
PRIMER_RIGHT_3_HAIRPIN_TH=32.64
PRIMER_INTERNAL_3_HAIRPIN_TH=44.20
PRIMER_LEFT_3_END_STABILITY=2.5700
PRIMER_RIGHT_3_END_STABILITY=2.6900
PRIMER_LEFT_4_PENALTY=1.819575
PRIMER_RIGHT_4_PENALTY=1.657646
PRIMER_LEFT_4_SEQUENCE=atggacatggatgccatcaa
PRIMER_RIGHT_4_SEQUENCE=tcattgatggcatccatgtcc
PRIMER_LEFT_4=9,20
PRIMER_RIGHT_4=31,21
PRIMER_LEFT_4_TM=58.180
PRIMER_RIGHT_4_TM=59.342
PRIMER_LEFT_4_GC_PERCENT=45.000
PRIMER_RIGHT_4_GC_PERCENT=47.619
PRIMER_LEFT_4_SELF_ANY_TH=7.69
PRIMER_RIGHT_4_SELF_ANY_TH=24.48
PRIMER_LEFT_4_SELF_END_TH=0.00
PRIMER_RIGHT_4_SELF_END_TH=0.00
PRIMER_LEFT_4_HAIRPIN_TH=44.12
PRIMER_RIGHT_4_HAIRPIN_TH=46.18
PRIMER_LEFT_4_END_STABILITY=2.5700
PRIMER_RIGHT_4_END_STABILITY=4.0200
PRIMER_LEFT_5_PENALTY=2.024122
PRIMER_RIGHT_5_PENALTY=1.657646
PRIMER_LEFT_5_SEQUENCE=gatttgatggacatggatgcca
PRIMER_RIGHT_5_SEQUENCE=ttgatggcatccatgtccatc
PRIMER_LEFT_5=3,22
PRIMER_RIGHT_5=28,21
PRIMER_LEFT_5_TM=59.976
PRIMER_RIGHT_5_TM=59.342
PRIMER_LEFT_5_GC_PERCENT=45.455
PRIMER_RIGHT_5_GC_PERCENT=47.619
PRIMER_LEFT_5_SELF_ANY_TH=0.00
PRIMER_RIGHT_5_SELF_ANY_TH=24.48
PRIMER_LEFT_5_SELF_END_TH=0.00
PRIMER_RIGHT_5_SELF_END_TH=23.68
PRIMER_LEFT_5_HAIRPIN_TH=38.48
PRIMER_RIGHT_5_HAIRPIN_TH=46.18
PRIMER_LEFT_5_END_STABILITY=4.9200
PRIMER_RIGHT_5_END_STABILITY=3.5100
PRIMER_LEFT_6_PENALTY=2.024122
PRIMER_RIGHT_6_PENALTY=1.657646
PRIMER_LEFT_6_SEQUENCE=tggatttgatggacatggatgc
PRIMER_RIGHT_6_SEQUENCE=gatggcatccatgtccatcaa
PRIMER_LEFT_6=1,22
PRIMER_RIGHT_6=26,21
PRIMER_LEFT_6_TM=59.976
PRIMER_RIGHT_6_TM=59.342
PRIMER_LEFT_6_GC_PERCENT=45.455
PRIMER_RIGHT_6_GC_PERCENT=47.619
PRIMER_LEFT_6_SELF_ANY_TH=0.00
PRIMER_RIGHT_6_SELF_ANY_TH=24.61
PRIMER_LEFT_6_SELF_END_TH=0.00
PRIMER_RIGHT_6_SELF_END_TH=0.00
PRIMER_LEFT_6_HAIRPIN_TH=0.00
PRIMER_RIGHT_6_HAIRPIN_TH=46.18
PRIMER_LEFT_6_END_STABILITY=3.9100
PRIMER_RIGHT_6_END_STABILITY=2.5700
PRIMER_LEFT_7_PENALTY=2.041530
PRIMER_RIGHT_7_PENALTY=1.819575
PRIMER_LEFT_7_SEQUENCE=atggacatggatgccatcaatg
PRIMER_RIGHT_7_SEQUENCE=attgatggcatccatgtcca
PRIMER_LEFT_7=9,22
PRIMER_RIGHT_7=29,20
PRIMER_LEFT_7_TM=60.042
PRIMER_RIGHT_7_TM=58.180
PRIMER_LEFT_7_GC_PERCENT=45.455
PRIMER_RIGHT_7_GC_PERCENT=45.000
PRIMER_LEFT_7_SELF_ANY_TH=19.55
PRIMER_RIGHT_7_SELF_ANY_TH=24.48
PRIMER_LEFT_7_SELF_END_TH=19.55
PRIMER_RIGHT_7_SELF_END_TH=2.36
PRIMER_LEFT_7_HAIRPIN_TH=44.12
PRIMER_RIGHT_7_HAIRPIN_TH=46.18
PRIMER_LEFT_7_END_STABILITY=2.8200
PRIMER_RIGHT_7_END_STABILITY=4.0200
PRIMER_LEFT_8_PENALTY=2.090718
PRIMER_RIGHT_8_PENALTY=1.819575
PRIMER_LEFT_8_SEQUENCE=atttgatggacatggatgcca
PRIMER_RIGHT_8_SEQUENCE=ttgatggcatccatgtccat
PRIMER_LEFT_8=4,21
PRIMER_RIGHT_8=28,20
PRIMER_LEFT_8_TM=58.909
PRIMER_RIGHT_8_TM=58.180
PRIMER_LEFT_8_GC_PERCENT=42.857
PRIMER_RIGHT_8_GC_PERCENT=45.000
PRIMER_LEFT_8_SELF_ANY_TH=0.00
PRIMER_RIGHT_8_SELF_ANY_TH=24.48
PRIMER_LEFT_8_SELF_END_TH=0.00
PRIMER_RIGHT_8_SELF_END_TH=13.31
PRIMER_LEFT_8_HAIRPIN_TH=38.48
PRIMER_RIGHT_8_HAIRPIN_TH=46.18
PRIMER_LEFT_8_END_STABILITY=4.9200
PRIMER_RIGHT_8_END_STABILITY=3.4100
PRIMER_LEFT_9_PENALTY=2.189272
PRIMER_RIGHT_9_PENALTY=1.819575
PRIMER_LEFT_9_SEQUENCE=ggacatggatgccatcaatg
PRIMER_RIGHT_9_SEQUENCE=atggcatccatgtccatcaa
PRIMER_LEFT_9=11,20
PRIMER_RIGHT_9=25,20
PRIMER_LEFT_9_TM=57.811
PRIMER_RIGHT_9_TM=58.180
PRIMER_LEFT_9_GC_PERCENT=50.000
PRIMER_RIGHT_9_GC_PERCENT=45.000
PRIMER_LEFT_9_SELF_ANY_TH=19.55
PRIMER_RIGHT_9_SELF_ANY_TH=0.00
PRIMER_LEFT_9_SELF_END_TH=19.55
PRIMER_RIGHT_9_SELF_END_TH=0.00
PRIMER_LEFT_9_HAIRPIN_TH=44.12
PRIMER_RIGHT_9_HAIRPIN_TH=44.27
PRIMER_LEFT_9_END_STABILITY=2.8200
PRIMER_RIGHT_9_END_STABILITY=2.5700
PRIMER_LEFT_10_PENALTY=2.306792
PRIMER_RIGHT_10_PENALTY=2.024122
PRIMER_LEFT_10_SEQUENCE=atggacatggatgccatcaat
PRIMER_RIGHT_10_SEQUENCE=gatggcatccatgtccatcaaa
PRIMER_LEFT_10=9,21
PRIMER_RIGHT_10=26,22
PRIMER_LEFT_10_TM=58.693
PRIMER_RIGHT_10_TM=59.976
PRIMER_LEFT_10_GC_PERCENT=42.857
PRIMER_RIGHT_10_GC_PERCENT=45.455
PRIMER_LEFT_10_SELF_ANY_TH=7.69
PRIMER_RIGHT_10_SELF_ANY_TH=24.61
PRIMER_LEFT_10_SELF_END_TH=0.00
PRIMER_RIGHT_10_SELF_END_TH=0.00
PRIMER_LEFT_10_HAIRPIN_TH=44.12
PRIMER_RIGHT_10_HAIRPIN_TH=46.18
PRIMER_LEFT_10_END_STABILITY=2.5700
PRIMER_RIGHT_10_END_STABILITY=2.6900
PRIMER_LEFT_11_PENALTY=2.632409
PRIMER_RIGHT_11_PENALTY=2.024122
PRIMER_LEFT_11_SEQUENCE=ggatttgatggacatggatgcc
PRIMER_RIGHT_11_SEQUENCE=tggcatccatgtccatcaaatc
PRIMER_LEFT_11=2,22
PRIMER_RIGHT_11=24,22
PRIMER_LEFT_11_TM=60.632
PRIMER_RIGHT_11_TM=59.976
PRIMER_LEFT_11_GC_PERCENT=50.000
PRIMER_RIGHT_11_GC_PERCENT=45.455
PRIMER_LEFT_11_SELF_ANY_TH=0.00
PRIMER_RIGHT_11_SELF_ANY_TH=0.00
PRIMER_LEFT_11_SELF_END_TH=0.00
PRIMER_RIGHT_11_SELF_END_TH=0.00
PRIMER_LEFT_11_HAIRPIN_TH=0.00
PRIMER_RIGHT_11_HAIRPIN_TH=32.64
PRIMER_LEFT_11_END_STABILITY=4.4000
PRIMER_RIGHT_11_END_STABILITY=2.1700
PRIMER_LEFT_12_PENALTY=2.738369
PRIMER_RIGHT_12_PENALTY=2.024122
PRIMER_LEFT_12_SEQUENCE=gatttgatggacatggatgcc
PRIMER_RIGHT_12_SEQUENCE=gcatccatgtccatcaaatcca
PRIMER_LEFT_12=3,21
PRIMER_RIGHT_12=22,22
PRIMER_LEFT_12_TM=58.262
PRIMER_RIGHT_12_TM=59.976
PRIMER_LEFT_12_GC_PERCENT=47.619
PRIMER_RIGHT_12_GC_PERCENT=45.455
PRIMER_LEFT_12_SELF_ANY_TH=0.00
PRIMER_RIGHT_12_SELF_ANY_TH=0.00
PRIMER_LEFT_12_SELF_END_TH=0.00
PRIMER_RIGHT_12_SELF_END_TH=0.00
PRIMER_LEFT_12_HAIRPIN_TH=0.00
PRIMER_RIGHT_12_HAIRPIN_TH=0.00
PRIMER_LEFT_12_END_STABILITY=4.4000
PRIMER_RIGHT_12_END_STABILITY=3.4100
PRIMER_LEFT_13_PENALTY=2.738369
PRIMER_RIGHT_13_PENALTY=2.041530
PRIMER_LEFT_13_SEQUENCE=ggatttgatggacatggatgc
PRIMER_RIGHT_13_SEQUENCE=cattgatggcatccatgtccat
PRIMER_LEFT_13=2,21
PRIMER_RIGHT_13=30,22
PRIMER_LEFT_13_TM=58.262
PRIMER_RIGHT_13_TM=60.042
PRIMER_LEFT_13_GC_PERCENT=47.619
PRIMER_RIGHT_13_GC_PERCENT=45.455
PRIMER_LEFT_13_SELF_ANY_TH=0.00
PRIMER_RIGHT_13_SELF_ANY_TH=24.48
PRIMER_LEFT_13_SELF_END_TH=0.00
PRIMER_RIGHT_13_SELF_END_TH=13.31
PRIMER_LEFT_13_HAIRPIN_TH=0.00
PRIMER_RIGHT_13_HAIRPIN_TH=46.18
PRIMER_LEFT_13_END_STABILITY=3.9100
PRIMER_RIGHT_13_END_STABILITY=3.4100
PRIMER_LEFT_14_PENALTY=2.954042
PRIMER_RIGHT_14_PENALTY=2.090718
PRIMER_LEFT_14_SEQUENCE=atttgatggacatggatgcc
PRIMER_RIGHT_14_SEQUENCE=atggcatccatgtccatcaaa
PRIMER_LEFT_14=4,20
PRIMER_RIGHT_14=25,21
PRIMER_LEFT_14_TM=57.046
PRIMER_RIGHT_14_TM=58.909
PRIMER_LEFT_14_GC_PERCENT=45.000
PRIMER_RIGHT_14_GC_PERCENT=42.857
PRIMER_LEFT_14_SELF_ANY_TH=0.00
PRIMER_RIGHT_14_SELF_ANY_TH=0.00
PRIMER_LEFT_14_SELF_END_TH=0.00
PRIMER_RIGHT_14_SELF_END_TH=0.00
PRIMER_LEFT_14_HAIRPIN_TH=0.00
PRIMER_RIGHT_14_HAIRPIN_TH=44.27
PRIMER_LEFT_14_END_STABILITY=4.4000
PRIMER_RIGHT_14_END_STABILITY=2.6900
PRIMER_LEFT_15_PENALTY=3.043060
PRIMER_RIGHT_15_PENALTY=2.090718
PRIMER_LEFT_15_SEQUENCE=tggacatggatgccatcaatga
PRIMER_RIGHT_15_SEQUENCE=tggcatccatgtccatcaaat
PRIMER_LEFT_15=10,22
PRIMER_RIGHT_15=24,21
PRIMER_LEFT_15_TM=61.043
PRIMER_RIGHT_15_TM=58.909
PRIMER_LEFT_15_GC_PERCENT=45.455
PRIMER_RIGHT_15_GC_PERCENT=42.857
PRIMER_LEFT_15_SELF_ANY_TH=26.38
PRIMER_RIGHT_15_SELF_ANY_TH=0.00
PRIMER_LEFT_15_SELF_END_TH=4.73
PRIMER_RIGHT_15_SELF_END_TH=0.00
PRIMER_LEFT_15_HAIRPIN_TH=44.12
PRIMER_RIGHT_15_HAIRPIN_TH=32.64
PRIMER_LEFT_15_END_STABILITY=2.5700
PRIMER_RIGHT_15_END_STABILITY=2.3200
PRIMER_LEFT_16_PENALTY=3.365788
PRIMER_RIGHT_16_PENALTY=2.189272
PRIMER_LEFT_16_SEQUENCE=atggatttgatggacatggatgc
PRIMER_RIGHT_16_SEQUENCE=cattgatggcatccatgtcc
PRIMER_LEFT_16=0,23
PRIMER_RIGHT_16=30,20
PRIMER_LEFT_16_TM=60.366
PRIMER_RIGHT_16_TM=57.811
PRIMER_LEFT_16_GC_PERCENT=43.478
PRIMER_RIGHT_16_GC_PERCENT=50.000
PRIMER_LEFT_16_SELF_ANY_TH=0.00
PRIMER_RIGHT_16_SELF_ANY_TH=24.48
PRIMER_LEFT_16_SELF_END_TH=0.00
PRIMER_RIGHT_16_SELF_END_TH=0.00
PRIMER_LEFT_16_HAIRPIN_TH=0.00
PRIMER_RIGHT_16_HAIRPIN_TH=46.18
PRIMER_LEFT_16_END_STABILITY=3.9100
PRIMER_RIGHT_16_END_STABILITY=4.0200
PRIMER_LEFT_17_PENALTY=3.382273
PRIMER_RIGHT_17_PENALTY=2.223627
PRIMER_LEFT_17_SEQUENCE=tggacatggatgccatcaa
PRIMER_RIGHT_17_SEQUENCE=attgatggcatccatgtccatc
PRIMER_LEFT_17=10,19
PRIMER_RIGHT_17=29,22
PRIMER_LEFT_17_TM=57.618
PRIMER_RIGHT_17_TM=59.776
PRIMER_LEFT_17_GC_PERCENT=47.368
PRIMER_RIGHT_17_GC_PERCENT=45.455
PRIMER_LEFT_17_SELF_ANY_TH=7.69
PRIMER_RIGHT_17_SELF_ANY_TH=24.48
PRIMER_LEFT_17_SELF_END_TH=0.00
PRIMER_RIGHT_17_SELF_END_TH=23.68
PRIMER_LEFT_17_HAIRPIN_TH=44.12
PRIMER_RIGHT_17_HAIRPIN_TH=46.18
PRIMER_LEFT_17_END_STABILITY=2.5700
PRIMER_RIGHT_17_END_STABILITY=3.5100
PRIMER_LEFT_18_PENALTY=3.382273
PRIMER_RIGHT_18_PENALTY=2.306792
PRIMER_LEFT_18_SEQUENCE=ttgatggacatggatgcca
PRIMER_RIGHT_18_SEQUENCE=attgatggcatccatgtccat
PRIMER_LEFT_18=6,19
PRIMER_RIGHT_18=29,21
PRIMER_LEFT_18_TM=57.618
PRIMER_RIGHT_18_TM=58.693
PRIMER_LEFT_18_GC_PERCENT=47.368
PRIMER_RIGHT_18_GC_PERCENT=42.857
PRIMER_LEFT_18_SELF_ANY_TH=0.00
PRIMER_RIGHT_18_SELF_ANY_TH=24.48
PRIMER_LEFT_18_SELF_END_TH=0.00
PRIMER_RIGHT_18_SELF_END_TH=13.31
PRIMER_LEFT_18_HAIRPIN_TH=38.48
PRIMER_RIGHT_18_HAIRPIN_TH=46.18
PRIMER_LEFT_18_END_STABILITY=4.9200
PRIMER_RIGHT_18_END_STABILITY=3.4100
PRIMER_LEFT_19_PENALTY=3.625785
PRIMER_RIGHT_19_PENALTY=2.632337
PRIMER_LEFT_19_SEQUENCE=atggacatggatgccatca
PRIMER_RIGHT_19_SEQUENCE=atggcatccatgtccatcaaat
PRIMER_LEFT_19=9,19
PRIMER_RIGHT_19=25,22
PRIMER_LEFT_19_TM=57.374
PRIMER_RIGHT_19_TM=59.368
PRIMER_LEFT_19_GC_PERCENT=47.368
PRIMER_RIGHT_19_GC_PERCENT=40.909
PRIMER_LEFT_19_SELF_ANY_TH=7.69
PRIMER_RIGHT_19_SELF_ANY_TH=0.00
PRIMER_LEFT_19_SELF_END_TH=0.00
PRIMER_RIGHT_19_SELF_END_TH=0.00
PRIMER_LEFT_19_HAIRPIN_TH=44.12
PRIMER_RIGHT_19_HAIRPIN_TH=44.27
PRIMER_LEFT_19_END_STABILITY=3.0700
PRIMER_RIGHT_19_END_STABILITY=2.3200
=
SEQUENCE_ID=Make a list of a small range
SEQUENCE_TEMPLATE=atggatttgatggacatggatgccatcaatga
PRIMER_LOWERCASE_MASKING=0
PRIMER_TASK=pick_primer_list
PRIMER_PICK_RIGHT_PRIMER=1
PRIMER_PICK_INTERNAL_OLIGO=0
PRIMER_PICK_LEFT_PRIMER=0
PRIMER_NUM_RETURN=2
PRIMER_WARNING=PRIMER_SALT_DIVALENT > 0.0 but PRIMER_DNTP_CONC <= 0.0; use reasonable value for PRIMER_DNTP_CONC
PRIMER_LEFT_NUM_RETURNED=0
PRIMER_RIGHT_NUM_RETURNED=2
PRIMER_INTERNAL_NUM_RETURNED=0
PRIMER_PAIR_NUM_RETURNED=0
PRIMER_RIGHT_0_PENALTY=1.354059
PRIMER_RIGHT_0_SEQUENCE=tgatggcatccatgtccatc
PRIMER_RIGHT_0=27,20
PRIMER_RIGHT_0_TM=58.646
PRIMER_RIGHT_0_GC_PERCENT=50.000
PRIMER_RIGHT_0_SELF_ANY_TH=24.48
PRIMER_RIGHT_0_SELF_END_TH=23.68
PRIMER_RIGHT_0_HAIRPIN_TH=46.18
PRIMER_RIGHT_0_END_STABILITY=3.5100
PRIMER_RIGHT_1_PENALTY=1.354059
PRIMER_RIGHT_1_SEQUENCE=gatggcatccatgtccatca
PRIMER_RIGHT_1=26,20
PRIMER_RIGHT_1_TM=58.646
PRIMER_RIGHT_1_GC_PERCENT=50.000
PRIMER_RIGHT_1_SELF_ANY_TH=24.61
PRIMER_RIGHT_1_SELF_END_TH=4.66
PRIMER_RIGHT_1_HAIRPIN_TH=46.18
PRIMER_RIGHT_1_END_STABILITY=3.0700
=
SEQUENCE_ID=Check left only too small
SEQUENCE_PRIMER=gtcctcgaaca
PRIMER_TASK=check_primers
PRIMER_PICK_RIGHT_PRIMER=0
PRIMER_PICK_INTERNAL_OLIGO=0
PRIMER_PICK_LEFT_PRIMER=1
PRIMER_PICK_ANYWAY=1
PRIMER_WARNING=Specified left primer < PRIMER_MIN_SIZE; PRIMER_SALT_DIVALENT > 0.0 but PRIMER_DNTP_CONC <= 0.0; use reasonable value for PRIMER_DNTP_CONC
PRIMER_LEFT_NUM_RETURNED=1
PRIMER_RIGHT_NUM_RETURNED=0
PRIMER_INTERNAL_NUM_RETURNED=0
PRIMER_PAIR_NUM_RETURNED=0
PRIMER_LEFT_0_PENALTY=33.747070
PRIMER_LEFT_0_PROBLEMS= Temperature too low; Too short;
PRIMER_LEFT_0_SEQUENCE=gtcctcgaaca
PRIMER_LEFT_0=0,11
PRIMER_LEFT_0_TM=35.253
PRIMER_LEFT_0_GC_PERCENT=54.545
PRIMER_LEFT_0_SELF_ANY_TH=0.00
PRIMER_LEFT_0_SELF_END_TH=0.00
PRIMER_LEFT_0_HAIRPIN_TH=0.00
PRIMER_LEFT_0_END_STABILITY=3.1800
=
SEQUENCE_ID=Check left only too big
SEQUENCE_PRIMER=tgacctccacttctctggacacagctccctcaccaa
PRIMER_TASK=check_primers
PRIMER_PICK_RIGHT_PRIMER=0
PRIMER_PICK_INTERNAL_OLIGO=0
PRIMER_PICK_LEFT_PRIMER=1
PRIMER_PICK_ANYWAY=1
PRIMER_WARNING=Specified left primer > PRIMER_MAX_SIZE; PRIMER_SALT_DIVALENT > 0.0 but PRIMER_DNTP_CONC <= 0.0; use reasonable value for PRIMER_DNTP_CONC
PRIMER_LEFT_NUM_RETURNED=1
PRIMER_RIGHT_NUM_RETURNED=0
PRIMER_INTERNAL_NUM_RETURNED=0
PRIMER_PAIR_NUM_RETURNED=0
PRIMER_LEFT_0_PENALTY=31.449855
PRIMER_LEFT_0_PROBLEMS= Temperature too high; Too long;
PRIMER_LEFT_0_SEQUENCE=tgacctccacttctctggacacagctccctcaccaa
PRIMER_LEFT_0=0,36
PRIMER_LEFT_0_TM=75.450
PRIMER_LEFT_0_GC_PERCENT=55.556
PRIMER_LEFT_0_SELF_ANY_TH=0.00
PRIMER_LEFT_0_SELF_END_TH=0.00
PRIMER_LEFT_0_HAIRPIN_TH=45.07
PRIMER_LEFT_0_END_STABILITY=3.6700
=
SEQUENCE_ID=Pick sequencing primers no target
SEQUENCE_TEMPLATE=atggatttgatggacatggatgccatcaatgacctccacttctctggacacagctccctcaccaacacccctacttccgacgaggattatggttgcacctggaaccactggtctcccatcgtcaactgggacacctttacaggtgccccagatgactttcaccacctcatggacaccatcatcgaggacagaacaactgtcctcgaacagctcagtccatctatcaccacaaccaccaccaccaccacaaccacggatgaggaagaagaagaaatggaaacaacaaccacaacaacaacaacggcaataaaaacgcatgaagttggtgatgattcaaaagggctaaaactagtgcacctattgatggctggcgcggaagccttaaccggttcaaccaaaaaccgtgacttagctcgagtgatattgatccggctcaaggaattagtctcacaacatgctaacggctccaacatggagagacttgccgctcatttcaccgaagcccttcatggactactagaaggtgctgggggtgcgcataataaccaccatcaccacaacaacaacaaacattacctcaccacaaatggtccccacgacaaccaaaacgacacacttgctgctttccaactacttcaagacatgtcaccttacgtcaaattcggccacttcacagccaatcaagccatcatcgaagccgtggcccacgaacgccgcgtccatgtcatcgactacgatatcatggaaggggtccaatgggcctcattaatccaatcacttgcttccaacaacaacggtccacatcttcggataaccgcattatcgcggacaggaaccggccggaggtcaatcgccaccgtacaagaaaccggcaggcgattaacctcctttgccgcttccctcggacaaccattttcttttcatcactgcagattagactccgatgaaactttccgtccttctgcattaaagcttgtacgtggagaggctttggttttcaactgcatgctgaatttacctcaccttagttaccgtgcaccggaatcagttgcttcgtttttaaacggagcaaaaacgttgaatccaaagcttgtgacgttggttgaagaagaagttggttctgttattggtgggtttgtggaaaggttcatggactcacttcatcattattcagcggtttttgattcattggaggctggttttccgatgcagaaccgggcccggactttggtggagagggtattttttgggcctagaattgcaggctcgttgggccggatatatagaacgggtggtgaagaggagagaaggtcatggggggagtggttaggtgaggtagggtttaggggagttccggtgagctttgcaaatcattgtcaagcaaagctgttgttagggctttttaatgatgggtatagggttgaagaggtgggtgtgggtagtaataagttggtgttggattggaaatcaagacgtttgctttctgcttccctttggacttgttcttcttcagattctgatttatag
PRIMER_TASK=pick_sequencing_primers
PRIMER_PICK_RIGHT_PRIMER=1
PRIMER_PICK_INTERNAL_OLIGO=0
PRIMER_PICK_LEFT_PRIMER=1
PRIMER_NUM_RETURN=50
PRIMER_WARNING=PRIMER_SALT_DIVALENT > 0.0 but PRIMER_DNTP_CONC <= 0.0; use reasonable value for PRIMER_DNTP_CONC
PRIMER_LEFT_NUM_RETURNED=4
PRIMER_RIGHT_NUM_RETURNED=4
PRIMER_INTERNAL_NUM_RETURNED=0
PRIMER_PAIR_NUM_RETURNED=0
PRIMER_LEFT_0_PENALTY=0.692905
PRIMER_RIGHT_0_PENALTY=0.098220
PRIMER_LEFT_0_SEQUENCE=ggatgccatcaatgacctcc
PRIMER_RIGHT_0_SEQUENCE=tgtcctcgatgatggtgtcc
PRIMER_LEFT_0=17,20
PRIMER_RIGHT_0=189,20
PRIMER_LEFT_0_TM=59.307
PRIMER_RIGHT_0_TM=60.098
PRIMER_LEFT_0_GC_PERCENT=55.000
PRIMER_RIGHT_0_GC_PERCENT=55.000
PRIMER_LEFT_0_SELF_ANY_TH=7.69
PRIMER_RIGHT_0_SELF_ANY_TH=0.00
PRIMER_LEFT_0_SELF_END_TH=0.00
PRIMER_RIGHT_0_SELF_END_TH=0.00
PRIMER_LEFT_0_HAIRPIN_TH=0.00
PRIMER_RIGHT_0_HAIRPIN_TH=0.00
PRIMER_LEFT_0_END_STABILITY=4.3000
PRIMER_RIGHT_0_END_STABILITY=4.0200
PRIMER_LEFT_1_PENALTY=1.038565
PRIMER_RIGHT_1_PENALTY=0.259988
PRIMER_LEFT_1_SEQUENCE=agggctaaaactagtgcacct
PRIMER_RIGHT_1_SEQUENCE=tgatggcttgattggctgtg
PRIMER_LEFT_1=338,21
PRIMER_RIGHT_1=690,20
PRIMER_LEFT_1_TM=59.961
PRIMER_RIGHT_1_TM=59.740
PRIMER_LEFT_1_GC_PERCENT=47.619
PRIMER_RIGHT_1_GC_PERCENT=50.000
PRIMER_LEFT_1_SELF_ANY_TH=15.14
PRIMER_RIGHT_1_SELF_ANY_TH=0.00
PRIMER_LEFT_1_SELF_END_TH=3.98
PRIMER_RIGHT_1_SELF_END_TH=0.00
PRIMER_LEFT_1_HAIRPIN_TH=0.00
PRIMER_RIGHT_1_HAIRPIN_TH=0.00
PRIMER_LEFT_1_END_STABILITY=4.0000
PRIMER_RIGHT_1_END_STABILITY=3.6600
PRIMER_LEFT_2_PENALTY=0.309295
PRIMER_RIGHT_2_PENALTY=0.028504
PRIMER_LEFT_2_SEQUENCE=gcattatcgcggacaggaac
PRIMER_RIGHT_2_SEQUENCE=ttctgcatcggaaaaccagc
PRIMER_LEFT_2=816,20
PRIMER_RIGHT_2=1213,20
PRIMER_LEFT_2_TM=60.309
PRIMER_RIGHT_2_TM=60.029
PRIMER_LEFT_2_GC_PERCENT=55.000
PRIMER_RIGHT_2_GC_PERCENT=50.000
PRIMER_LEFT_2_SELF_ANY_TH=6.47
PRIMER_RIGHT_2_SELF_ANY_TH=0.00
PRIMER_LEFT_2_SELF_END_TH=0.00
PRIMER_RIGHT_2_SELF_END_TH=0.00
PRIMER_LEFT_2_HAIRPIN_TH=0.00
PRIMER_RIGHT_2_HAIRPIN_TH=0.00
PRIMER_LEFT_2_END_STABILITY=3.6200
PRIMER_RIGHT_2_END_STABILITY=4.8500
PRIMER_LEFT_3_PENALTY=0.270229
PRIMER_RIGHT_3_PENALTY=3.267431
PRIMER_LEFT_3_SEQUENCE=ggtgaggtagggtttagggg
PRIMER_RIGHT_3_SEQUENCE=tgaagaagaacaagtccaaaggg
PRIMER_LEFT_3=1329,20
PRIMER_RIGHT_3=1511,23
PRIMER_LEFT_3_TM=59.730
PRIMER_RIGHT_3_TM=59.733
PRIMER_LEFT_3_GC_PERCENT=60.000
PRIMER_RIGHT_3_GC_PERCENT=43.478
PRIMER_LEFT_3_SELF_ANY_TH=0.00
PRIMER_RIGHT_3_SELF_ANY_TH=0.00
PRIMER_LEFT_3_SELF_END_TH=0.00
PRIMER_RIGHT_3_SELF_END_TH=0.00
PRIMER_LEFT_3_HAIRPIN_TH=0.00
PRIMER_RIGHT_3_HAIRPIN_TH=0.00
PRIMER_LEFT_3_END_STABILITY=4.7900
PRIMER_RIGHT_3_END_STABILITY=3.9500
=
SEQUENCE_ID=Pick sequencing primers small target
SEQUENCE_TEMPLATE=atggatttgatggacatggatgccatcaatgacctccacttctctggacacagctccctcaccaacacccctacttccgacgaggattatggttgcacctggaaccactggtctcccatcgtcaactgggacacctttacaggtgccccagatgactttcaccacctcatggacaccatcatcgaggacagaacaactgtcctcgaacagctcagtccatctatcaccacaaccaccaccaccaccacaaccacggatgaggaagaagaagaaatggaaacaacaaccacaacaacaacaacggcaataaaaacgcatgaagttggtgatgattcaaaagggctaaaactagtgcacctattgatggctggcgcggaagccttaaccggttcaaccaaaaaccgtgacttagctcgagtgatattgatccggctcaaggaattagtctcacaacatgctaacggctccaacatggagagacttgccgctcatttcaccgaagcccttcatggactactagaaggtgctgggggtgcgcataataaccaccatcaccacaacaacaacaaacattacctcaccacaaatggtccccacgacaaccaaaacgacacacttgctgctttccaactacttcaagacatgtcaccttacgtcaaattcggccacttcacagccaatcaagccatcatcgaagccgtggcccacgaacgccgcgtccatgtcatcgactacgatatcatggaaggggtccaatgggcctcattaatccaatcacttgcttccaacaacaacggtccacatcttcggataaccgcattatcgcggacaggaaccggccggaggtcaatcgccaccgtacaagaaaccggcaggcgattaacctcctttgccgcttccctcggacaaccattttcttttcatcactgcagattagactccgatgaaactttccgtccttctgcattaaagcttgtacgtggagaggctttggttttcaactgcatgctgaatttacctcaccttagttaccgtgcaccggaatcagttgcttcgtttttaaacggagcaaaaacgttgaatccaaagcttgtgacgttggttgaagaagaagttggttctgttattggtgggtttgtggaaaggttcatggactcacttcatcattattcagcggtttttgattcattggaggctggttttccgatgcagaaccgggcccggactttggtggagagggtattttttgggcctagaattgcaggctcgttgggccggatatatagaacgggtggtgaagaggagagaaggtcatggggggagtggttaggtgaggtagggtttaggggagttccggtgagctttgcaaatcattgtcaagcaaagctgttgttagggctttttaatgatgggtatagggttgaagaggtgggtgtgggtagtaataagttggtgttggattggaaatcaagacgtttgctttctgcttccctttggacttgttcttcttcagattctgatttatag
PRIMER_TASK=pick_sequencing_primers
PRIMER_PICK_RIGHT_PRIMER=1
PRIMER_PICK_INTERNAL_OLIGO=0
PRIMER_PICK_LEFT_PRIMER=1
SEQUENCE_TARGET=450,37
PRIMER_NUM_RETURN=50
PRIMER_WARNING=PRIMER_SALT_DIVALENT > 0.0 but PRIMER_DNTP_CONC <= 0.0; use reasonable value for PRIMER_DNTP_CONC
PRIMER_LEFT_NUM_RETURNED=1
PRIMER_RIGHT_NUM_RETURNED=1
PRIMER_INTERNAL_NUM_RETURNED=0
PRIMER_PAIR_NUM_RETURNED=0
PRIMER_LEFT_0_PENALTY=0.808071
PRIMER_RIGHT_0_PENALTY=0.029777
PRIMER_LEFT_0_SEQUENCE=cacaacaacaacaacggcaa
PRIMER_RIGHT_0_SEQUENCE=gaagtggccgaatttgacgt
PRIMER_LEFT_0=287,20
PRIMER_RIGHT_0=671,20
PRIMER_LEFT_0_TM=59.192
PRIMER_RIGHT_0_TM=60.030
PRIMER_LEFT_0_GC_PERCENT=45.000
PRIMER_RIGHT_0_GC_PERCENT=50.000
PRIMER_LEFT_0_SELF_ANY_TH=0.00
PRIMER_RIGHT_0_SELF_ANY_TH=0.00
PRIMER_LEFT_0_SELF_END_TH=0.00
PRIMER_RIGHT_0_SELF_END_TH=0.00
PRIMER_LEFT_0_HAIRPIN_TH=0.00
PRIMER_RIGHT_0_HAIRPIN_TH=0.00
PRIMER_LEFT_0_END_STABILITY=4.5200
PRIMER_RIGHT_0_END_STABILITY=4.3400
=
SEQUENCE_ID=Pick sequencing primers left only
SEQUENCE_TEMPLATE=atggatttgatggacatggatgccatcaatgacctccacttctctggacacagctccctcaccaacacccctacttccgacgaggattatggttgcacctggaaccactggtctcccatcgtcaactgggacacctttacaggtgccccagatgactttcaccacctcatggacaccatcatcgaggacagaacaactgtcctcgaacagctcagtccatctatcaccacaaccaccaccaccaccacaaccacggatgaggaagaagaagaaatggaaacaacaaccacaacaacaacaacggcaataaaaacgcatgaagttggtgatgattcaaaagggctaaaactagtgcacctattgatggctggcgcggaagccttaaccggttcaaccaaaaaccgtgacttagctcgagtgatattgatccggctcaaggaattagtctcacaacatgctaacggctccaacatggagagacttgccgctcatttcaccgaagcccttcatggactactagaaggtgctgggggtgcgcataataaccaccatcaccacaacaacaacaaacattacctcaccacaaatggtccccacgacaaccaaaacgacacacttgctgctttccaactacttcaagacatgtcaccttacgtcaaattcggccacttcacagccaatcaagccatcatcgaagccgtggcccacgaacgccgcgtccatgtcatcgactacgatatcatggaaggggtccaatgggcctcattaatccaatcacttgcttccaacaacaacggtccacatcttcggataaccgcattatcgcggacaggaaccggccggaggtcaatcgccaccgtacaagaaaccggcaggcgattaacctcctttgccgcttccctcggacaaccattttcttttcatcactgcagattagactccgatgaaactttccgtccttctgcattaaagcttgtacgtggagaggctttggttttcaactgcatgctgaatttacctcaccttagttaccgtgcaccggaatcagttgcttcgtttttaaacggagcaaaaacgttgaatccaaagcttgtgacgttggttgaagaagaagttggttctgttattggtgggtttgtggaaaggttcatggactcacttcatcattattcagcggtttttgattcattggaggctggttttccgatgcagaaccgggcccggactttggtggagagggtattttttgggcctagaattgcaggctcgttgggccggatatatagaacgggtggtgaagaggagagaaggtcatggggggagtggttaggtgaggtagggtttaggggagttccggtgagctttgcaaatcattgtcaagcaaagctgttgttagggctttttaatgatgggtatagggttgaagaggtgggtgtgggtagtaataagttggtgttggattggaaatcaagacgtttgctttctgcttccctttggacttgttcttcttcagattctgatttatag
PRIMER_TASK=pick_sequencing_primers
PRIMER_PICK_RIGHT_PRIMER=0
PRIMER_PICK_INTERNAL_OLIGO=0
PRIMER_PICK_LEFT_PRIMER=1
SEQUENCE_TARGET=150,800
PRIMER_NUM_RETURN=50
PRIMER_WARNING=PRIMER_SALT_DIVALENT > 0.0 but PRIMER_DNTP_CONC <= 0.0; use reasonable value for PRIMER_DNTP_CONC
PRIMER_LEFT_NUM_RETURNED=2
PRIMER_RIGHT_NUM_RETURNED=0
PRIMER_INTERNAL_NUM_RETURNED=0
PRIMER_PAIR_NUM_RETURNED=0
PRIMER_LEFT_0_PENALTY=0.692905
PRIMER_LEFT_0_SEQUENCE=ggatgccatcaatgacctcc
PRIMER_LEFT_0=17,20
PRIMER_LEFT_0_TM=59.307
PRIMER_LEFT_0_GC_PERCENT=55.000
PRIMER_LEFT_0_SELF_ANY_TH=7.69
PRIMER_LEFT_0_SELF_END_TH=0.00
PRIMER_LEFT_0_HAIRPIN_TH=0.00
PRIMER_LEFT_0_END_STABILITY=4.3000
PRIMER_LEFT_1_PENALTY=0.097666
PRIMER_LEFT_1_SEQUENCE=ccgaagcccttcatggacta
PRIMER_LEFT_1=496,20
PRIMER_LEFT_1_TM=60.098
PRIMER_LEFT_1_GC_PERCENT=55.000
PRIMER_LEFT_1_SELF_ANY_TH=9.08
PRIMER_LEFT_1_SELF_END_TH=0.00
PRIMER_LEFT_1_HAIRPIN_TH=40.00
PRIMER_LEFT_1_END_STABILITY=2.5900
=
SEQUENCE_ID=Pick sequencing primers right only
SEQUENCE_TEMPLATE=atggatttgatggacatggatgccatcaatgacctccacttctctggacacagctccctcaccaacacccctacttccgacgaggattatggttgcacctggaaccactggtctcccatcgtcaactgggacacctttacaggtgccccagatgactttcaccacctcatggacaccatcatcgaggacagaacaactgtcctcgaacagctcagtccatctatcaccacaaccaccaccaccaccacaaccacggatgaggaagaagaagaaatggaaacaacaaccacaacaacaacaacggcaataaaaacgcatgaagttggtgatgattcaaaagggctaaaactagtgcacctattgatggctggcgcggaagccttaaccggttcaaccaaaaaccgtgacttagctcgagtgatattgatccggctcaaggaattagtctcacaacatgctaacggctccaacatggagagacttgccgctcatttcaccgaagcccttcatggactactagaaggtgctgggggtgcgcataataaccaccatcaccacaacaacaacaaacattacctcaccacaaatggtccccacgacaaccaaaacgacacacttgctgctttccaactacttcaagacatgtcaccttacgtcaaattcggccacttcacagccaatcaagccatcatcgaagccgtggcccacgaacgccgcgtccatgtcatcgactacgatatcatggaaggggtccaatgggcctcattaatccaatcacttgcttccaacaacaacggtccacatcttcggataaccgcattatcgcggacaggaaccggccggaggtcaatcgccaccgtacaagaaaccggcaggcgattaacctcctttgccgcttccctcggacaaccattttcttttcatcactgcagattagactccgatgaaactttccgtccttctgcattaaagcttgtacgtggagaggctttggttttcaactgcatgctgaatttacctcaccttagttaccgtgcaccggaatcagttgcttcgtttttaaacggagcaaaaacgttgaatccaaagcttgtgacgttggttgaagaagaagttggttctgttattggtgggtttgtggaaaggttcatggactcacttcatcattattcagcggtttttgattcattggaggctggttttccgatgcagaaccgggcccggactttggtggagagggtattttttgggcctagaattgcaggctcgttgggccggatatatagaacgggtggtgaagaggagagaaggtcatggggggagtggttaggtgaggtagggtttaggggagttccggtgagctttgcaaatcattgtcaagcaaagctgttgttagggctttttaatgatgggtatagggttgaagaggtgggtgtgggtagtaataagttggtgttggattggaaatcaagacgtttgctttctgcttccctttggacttgttcttcttcagattctgatttatag
PRIMER_TASK=pick_sequencing_primers
PRIMER_PICK_RIGHT_PRIMER=1
PRIMER_PICK_INTERNAL_OLIGO=0
PRIMER_PICK_LEFT_PRIMER=0
SEQUENCE_TARGET=150,800
PRIMER_NUM_RETURN=50
PRIMER_WARNING=PRIMER_SALT_DIVALENT > 0.0 but PRIMER_DNTP_CONC <= 0.0; use reasonable value for PRIMER_DNTP_CONC
PRIMER_LEFT_NUM_RETURNED=0
PRIMER_RIGHT_NUM_RETURNED=2
PRIMER_INTERNAL_NUM_RETURNED=0
PRIMER_PAIR_NUM_RETURNED=0
PRIMER_RIGHT_0_PENALTY=0.113686
PRIMER_RIGHT_0_SEQUENCE=tggaaagcagcaagtgtgtc
PRIMER_RIGHT_0=628,20
PRIMER_RIGHT_0_TM=59.886
PRIMER_RIGHT_0_GC_PERCENT=50.000
PRIMER_RIGHT_0_SELF_ANY_TH=0.00
PRIMER_RIGHT_0_SELF_END_TH=0.00
PRIMER_RIGHT_0_HAIRPIN_TH=0.00
PRIMER_RIGHT_0_END_STABILITY=3.6700
PRIMER_RIGHT_1_PENALTY=0.112929
PRIMER_RIGHT_1_SEQUENCE=ccaacgtcacaagctttgga
PRIMER_RIGHT_1=1101,20
PRIMER_RIGHT_1_TM=59.887
PRIMER_RIGHT_1_GC_PERCENT=50.000
PRIMER_RIGHT_1_SELF_ANY_TH=6.15
PRIMER_RIGHT_1_SELF_END_TH=0.00
PRIMER_RIGHT_1_HAIRPIN_TH=40.05
PRIMER_RIGHT_1_END_STABILITY=3.5300
=
SEQUENCE_ID=Pick sequencing primers three target
SEQUENCE_TEMPLATE=atggatttgatggacatggatgccatcaatgacctccacttctctggacacagctccctcaccaacacccctacttccgacgaggattatggttgcacctggaaccactggtctcccatcgtcaactgggacacctttacaggtgccccagatgactttcaccacctcatggacaccatcatcgaggacagaacaactgtcctcgaacagctcagtccatctatcaccacaaccaccaccaccaccacaaccacggatgaggaagaagaagaaatggaaacaacaaccacaacaacaacaacggcaataaaaacgcatgaagttggtgatgattcaaaagggctaaaactagtgcacctattgatggctggcgcggaagccttaaccggttcaaccaaaaaccgtgacttagctcgagtgatattgatccggctcaaggaattagtctcacaacatgctaacggctccaacatggagagacttgccgctcatttcaccgaagcccttcatggactactagaaggtgctgggggtgcgcataataaccaccatcaccacaacaacaacaaacattacctcaccacaaatggtccccacgacaaccaaaacgacacacttgctgctttccaactacttcaagacatgtcaccttacgtcaaattcggccacttcacagccaatcaagccatcatcgaagccgtggcccacgaacgccgcgtccatgtcatcgactacgatatcatggaaggggtccaatgggcctcattaatccaatcacttgcttccaacaacaacggtccacatcttcggataaccgcattatcgcggacaggaaccggccggaggtcaatcgccaccgtacaagaaaccggcaggcgattaacctcctttgccgcttccctcggacaaccattttcttttcatcactgcagattagactccgatgaaactttccgtccttctgcattaaagcttgtacgtggagaggctttggttttcaactgcatgctgaatttacctcaccttagttaccgtgcaccggaatcagttgcttcgtttttaaacggagcaaaaacgttgaatccaaagcttgtgacgttggttgaagaagaagttggttctgttattggtgggtttgtggaaaggttcatggactcacttcatcattattcagcggtttttgattcattggaggctggttttccgatgcagaaccgggcccggactttggtggagagggtattttttgggcctagaattgcaggctcgttgggccggatatatagaacgggtggtgaagaggagagaaggtcatggggggagtggttaggtgaggtagggtttaggggagttccggtgagctttgcaaatcattgtcaagcaaagctgttgttagggctttttaatgatgggtatagggttgaagaggtgggtgtgggtagtaataagttggtgttggattggaaatcaagacgtttgctttctgcttccctttggacttgttcttcttcagattctgatttatag
PRIMER_TASK=pick_sequencing_primers
PRIMER_PICK_RIGHT_PRIMER=1
PRIMER_PICK_INTERNAL_OLIGO=0
PRIMER_PICK_LEFT_PRIMER=1
SEQUENCE_TARGET=350,37 450,100 850,500 1200,100
PRIMER_NUM_RETURN=50
PRIMER_WARNING=PRIMER_SALT_DIVALENT > 0.0 but PRIMER_DNTP_CONC <= 0.0; use reasonable value for PRIMER_DNTP_CONC
PRIMER_LEFT_NUM_RETURNED=5
PRIMER_RIGHT_NUM_RETURNED=5
PRIMER_INTERNAL_NUM_RETURNED=0
PRIMER_PAIR_NUM_RETURNED=0
PRIMER_LEFT_0_PENALTY=0.098220
PRIMER_RIGHT_0_PENALTY=0.042322
PRIMER_LEFT_0_SEQUENCE=ggacaccatcatcgaggaca
PRIMER_RIGHT_0_SEQUENCE=gttattatgcgcacccccag
PRIMER_LEFT_0=170,20
PRIMER_RIGHT_0=545,20
PRIMER_LEFT_0_TM=60.098
PRIMER_RIGHT_0_TM=59.958
PRIMER_LEFT_0_GC_PERCENT=55.000
PRIMER_RIGHT_0_GC_PERCENT=55.000
PRIMER_LEFT_0_SELF_ANY_TH=0.71
PRIMER_RIGHT_0_SELF_ANY_TH=15.51
PRIMER_LEFT_0_SELF_END_TH=0.00
PRIMER_RIGHT_0_SELF_END_TH=0.00
PRIMER_LEFT_0_HAIRPIN_TH=0.00
PRIMER_RIGHT_0_HAIRPIN_TH=0.00
PRIMER_LEFT_0_END_STABILITY=4.0200
PRIMER_RIGHT_0_END_STABILITY=4.4500
PRIMER_LEFT_1_PENALTY=0.808071
PRIMER_RIGHT_1_PENALTY=0.044209
PRIMER_LEFT_1_SEQUENCE=cacaacaacaacaacggcaa
PRIMER_RIGHT_1_SEQUENCE=ttgattggctgtgaagtggc
PRIMER_LEFT_1=287,20
PRIMER_RIGHT_1=683,20
PRIMER_LEFT_1_TM=59.192
PRIMER_RIGHT_1_TM=59.956
PRIMER_LEFT_1_GC_PERCENT=45.000
PRIMER_RIGHT_1_GC_PERCENT=50.000
PRIMER_LEFT_1_SELF_ANY_TH=0.00
PRIMER_RIGHT_1_SELF_ANY_TH=0.00
PRIMER_LEFT_1_SELF_END_TH=0.00
PRIMER_RIGHT_1_SELF_END_TH=0.00
PRIMER_LEFT_1_HAIRPIN_TH=0.00
PRIMER_RIGHT_1_HAIRPIN_TH=0.00
PRIMER_LEFT_1_END_STABILITY=4.5200
PRIMER_RIGHT_1_END_STABILITY=5.0100
PRIMER_LEFT_2_PENALTY=0.029777
PRIMER_RIGHT_2_PENALTY=0.252674
PRIMER_LEFT_2_SEQUENCE=acgtcaaattcggccacttc
PRIMER_RIGHT_2_SEQUENCE=aacgaagcaactgattccgg
PRIMER_LEFT_2=652,20
PRIMER_RIGHT_2=1057,20
PRIMER_LEFT_2_TM=60.030
PRIMER_RIGHT_2_TM=59.747
PRIMER_LEFT_2_GC_PERCENT=50.000
PRIMER_RIGHT_2_GC_PERCENT=50.000
PRIMER_LEFT_2_SELF_ANY_TH=2.58
PRIMER_RIGHT_2_SELF_ANY_TH=0.00
PRIMER_LEFT_2_SELF_END_TH=0.00
PRIMER_RIGHT_2_SELF_END_TH=0.00
PRIMER_LEFT_2_HAIRPIN_TH=0.00
PRIMER_RIGHT_2_HAIRPIN_TH=37.93
PRIMER_LEFT_2_END_STABILITY=3.0100
PRIMER_RIGHT_2_END_STABILITY=5.1400
PRIMER_LEFT_3_PENALTY=2.108988
PRIMER_RIGHT_3_PENALTY=3.267431
PRIMER_LEFT_3_SEQUENCE=gtggaaaggttcatggactcac
PRIMER_RIGHT_3_SEQUENCE=tgaagaagaacaagtccaaaggg
PRIMER_LEFT_3=1137,22
PRIMER_RIGHT_3=1511,23
PRIMER_LEFT_3_TM=60.109
PRIMER_RIGHT_3_TM=59.733
PRIMER_LEFT_3_GC_PERCENT=50.000
PRIMER_RIGHT_3_GC_PERCENT=43.478
PRIMER_LEFT_3_SELF_ANY_TH=4.28
PRIMER_RIGHT_3_SELF_ANY_TH=0.00
PRIMER_LEFT_3_SELF_END_TH=3.06
PRIMER_RIGHT_3_SELF_END_TH=0.00
PRIMER_LEFT_3_HAIRPIN_TH=0.00
PRIMER_RIGHT_3_HAIRPIN_TH=0.00
PRIMER_LEFT_3_END_STABILITY=3.5100
PRIMER_RIGHT_3_END_STABILITY=3.9500
PRIMER_LEFT_4_PENALTY=0.252674
PRIMER_RIGHT_4_PENALTY=0.169561
PRIMER_LEFT_4_SEQUENCE=ccggaatcagttgcttcgtt
PRIMER_RIGHT_4_SEQUENCE=tactacccacacccacctct
PRIMER_LEFT_4=1038,20
PRIMER_RIGHT_4=1443,20
PRIMER_LEFT_4_TM=59.747
PRIMER_RIGHT_4_TM=60.170
PRIMER_LEFT_4_GC_PERCENT=50.000
PRIMER_RIGHT_4_GC_PERCENT=55.000
PRIMER_LEFT_4_SELF_ANY_TH=0.00
PRIMER_RIGHT_4_SELF_ANY_TH=0.00
PRIMER_LEFT_4_SELF_END_TH=0.00
PRIMER_RIGHT_4_SELF_END_TH=0.00
PRIMER_LEFT_4_HAIRPIN_TH=0.00
PRIMER_RIGHT_4_HAIRPIN_TH=0.00
PRIMER_LEFT_4_END_STABILITY=3.8500
PRIMER_RIGHT_4_END_STABILITY=3.6900
=
SEQUENCE_ID=Pick fixed left primer
SEQUENCE_TEMPLATE=atggatttgatggacatggatgccatcaatgacctccacttctctggacacagctccctcaccaacacccctacttccgacgaggattatggttgcacctggaaccactggtctcccatcgtcaactgggacacctttacaggtgccccagatgactttcaccacctcatggacaccatcatcgaggacagaacaactgtcctcgaacagctcagtccatctatcaccacaaccaccaccaccaccacaaccacggatgaggaagaagaagaaatggaaacaacaaccacaacaacaacaacggcaataaaaacgcatgaagttggtgatgattcaaaagggctaaaactagtgcacctattgatggctggcgcggaagccttaaccggttcaaccaaaaaccgtgacttagctcgagtgatattgatccggctcaaggaattagtctcacaacatgctaacggctccaacatggagagacttgccgctcatttcaccgaagcccttcatggactactagaaggtgctgggggtgcgcataataaccaccatcaccacaacaacaacaaacattacctcaccacaaatggtccccacgacaaccaaaacgacacacttgctgctttccaactacttcaagacatgtcaccttacgtcaaattcggccacttcacagccaatcaagccatcatcgaagccgtggcccacgaacgccgcgtccatgtcatcgactacgatatcatggaaggggtccaatgggcctcattaatccaatcacttgcttccaacaacaacggtccacatcttcggataaccgcattatcgcggacaggaaccggccggaggtcaatcgccaccgtacaagaaaccggcaggcgattaacctcctttgccgcttccctcggacaaccattttcttttcatcactgcagattagactccgatgaaactttccgtccttctgcattaaagcttgtacgtggagaggctttggttttcaactgcatgctgaatttacctcaccttagttaccgtgcaccggaatcagttgcttcgtttttaaacggagcaaaaacgttgaatccaaagcttgtgacgttggttgaagaagaagttggttctgttattggtgggtttgtggaaaggttcatggactcacttcatcattattcagcggtttttgattcattggaggctggttttccgatgcagaaccgggcccggactttggtggagagggtattttttgggcctagaattgcaggctcgttgggccggatatatagaacgggtggtgaagaggagagaaggtcatggggggagtggttaggtgaggtagggtttaggggagttccggtgagctttgcaaatcattgtcaagcaaagctgttgttagggctttttaatgatgggtatagggttgaagaggtgggtgtgggtagtaataagttggtgttggattggaaatcaagacgtttgctttctgcttccctttggacttgttcttcttcagattctgatttatag
PRIMER_TASK=pick_detection_primers
PRIMER_PICK_RIGHT_PRIMER=1
PRIMER_PICK_INTERNAL_OLIGO=0
PRIMER_PICK_LEFT_PRIMER=1
SEQUENCE_FORCE_LEFT_START=15
SEQUENCE_FORCE_LEFT_END=34
PRIMER_NUM_RETURN=3
PRIMER_PICK_ANYWAY=1
PRIMER_WARNING=PRIMER_SALT_DIVALENT > 0.0 but PRIMER_DNTP_CONC <= 0.0; use reasonable value for PRIMER_DNTP_CONC
PRIMER_LEFT_NUM_RETURNED=3
PRIMER_RIGHT_NUM_RETURNED=3
PRIMER_INTERNAL_NUM_RETURNED=0
PRIMER_PAIR_NUM_RETURNED=3
PRIMER_PAIR_0_PENALTY=2.220283
PRIMER_LEFT_0_PENALTY=2.122953
PRIMER_RIGHT_0_PENALTY=0.097330
PRIMER_LEFT_0_SEQUENCE=atggatgccatcaatgacct
PRIMER_RIGHT_0_SEQUENCE=aaaggtgtcccagttgacga
PRIMER_LEFT_0=15,20
PRIMER_RIGHT_0=137,20
PRIMER_LEFT_0_TM=57.877
PRIMER_RIGHT_0_TM=60.097
PRIMER_LEFT_0_GC_PERCENT=45.000
PRIMER_RIGHT_0_GC_PERCENT=50.000
PRIMER_LEFT_0_SELF_ANY_TH=7.69
PRIMER_RIGHT_0_SELF_ANY_TH=0.00
PRIMER_LEFT_0_SELF_END_TH=0.00
PRIMER_RIGHT_0_SELF_END_TH=0.00
PRIMER_LEFT_0_HAIRPIN_TH=41.92
PRIMER_RIGHT_0_HAIRPIN_TH=38.67
PRIMER_LEFT_0_END_STABILITY=3.8500
PRIMER_RIGHT_0_END_STABILITY=4.2000
PRIMER_PAIR_0_COMPL_ANY_TH=0.00
PRIMER_PAIR_0_COMPL_END_TH=0.00
PRIMER_PAIR_0_PRODUCT_SIZE=123
PRIMER_PAIR_0_PRODUCT_TM=86.9
PRIMER_PAIR_1_PENALTY=2.221173
PRIMER_LEFT_1_PENALTY=2.122953
PRIMER_RIGHT_1_PENALTY=0.098220
PRIMER_LEFT_1_SEQUENCE=atggatgccatcaatgacct
PRIMER_RIGHT_1_SEQUENCE=gtcctcgatgatggtgtcca
PRIMER_LEFT_1=15,20
PRIMER_RIGHT_1=188,20
PRIMER_LEFT_1_TM=57.877
PRIMER_RIGHT_1_TM=60.098
PRIMER_LEFT_1_GC_PERCENT=45.000
PRIMER_RIGHT_1_GC_PERCENT=55.000
PRIMER_LEFT_1_SELF_ANY_TH=7.69
PRIMER_RIGHT_1_SELF_ANY_TH=0.00
PRIMER_LEFT_1_SELF_END_TH=0.00
PRIMER_RIGHT_1_SELF_END_TH=0.00
PRIMER_LEFT_1_HAIRPIN_TH=41.92
PRIMER_RIGHT_1_HAIRPIN_TH=38.57
PRIMER_LEFT_1_END_STABILITY=3.8500
PRIMER_RIGHT_1_END_STABILITY=4.0200
PRIMER_PAIR_1_COMPL_ANY_TH=20.44
PRIMER_PAIR_1_COMPL_END_TH=20.44
PRIMER_PAIR_1_PRODUCT_SIZE=174
PRIMER_PAIR_1_PRODUCT_TM=88.5
PRIMER_PAIR_2_PENALTY=2.221173
PRIMER_LEFT_2_PENALTY=2.122953
PRIMER_RIGHT_2_PENALTY=0.098220
PRIMER_LEFT_2_SEQUENCE=atggatgccatcaatgacct
PRIMER_RIGHT_2_SEQUENCE=tgtcctcgatgatggtgtcc
PRIMER_LEFT_2=15,20
PRIMER_RIGHT_2=189,20
PRIMER_LEFT_2_TM=57.877
PRIMER_RIGHT_2_TM=60.098
PRIMER_LEFT_2_GC_PERCENT=45.000
PRIMER_RIGHT_2_GC_PERCENT=55.000
PRIMER_LEFT_2_SELF_ANY_TH=7.69
PRIMER_RIGHT_2_SELF_ANY_TH=0.00
PRIMER_LEFT_2_SELF_END_TH=0.00
PRIMER_RIGHT_2_SELF_END_TH=0.00
PRIMER_LEFT_2_HAIRPIN_TH=41.92
PRIMER_RIGHT_2_HAIRPIN_TH=0.00
PRIMER_LEFT_2_END_STABILITY=3.8500
PRIMER_RIGHT_2_END_STABILITY=4.0200
PRIMER_PAIR_2_COMPL_ANY_TH=15.21
PRIMER_PAIR_2_COMPL_END_TH=15.21
PRIMER_PAIR_2_PRODUCT_SIZE=175
PRIMER_PAIR_2_PRODUCT_TM=88.4
=
SEQUENCE_ID=Pick fixed right primer
SEQUENCE_TEMPLATE=atggatttgatggacatggatgccatcaatgacctccacttctctggacacagctccctcaccaacacccctacttccgacgaggattatggttgcacctggaaccactggtctcccatcgtcaactgggacacctttacaggtgccccagatgactttcaccacctcatggacaccatcatcgaggacagaacaactgtcctcgaacagctcagtccatctatcaccacaaccaccaccaccaccacaaccacggatgaggaagaagaagaaatggaaacaacaaccacaacaacaacaacggcaataaaaacgcatgaagttggtgatgattcaaaagggctaaaactagtgcacctattgatggctggcgcggaagccttaaccggttcaaccaaaaaccgtgacttagctcgagtgatattgatccggctcaaggaattagtctcacaacatgctaacggctccaacatggagagacttgccgctcatttcaccgaagcccttcatggactactagaaggtgctgggggtgcgcataataaccaccatcaccacaacaacaacaaacattacctcaccacaaatggtccccacgacaaccaaaacgacacacttgctgctttccaactacttcaagacatgtcaccttacgtcaaattcggccacttcacagccaatcaagccatcatcgaagccgtggcccacgaacgccgcgtccatgtcatcgactacgatatcatggaaggggtccaatgggcctcattaatccaatcacttgcttccaacaacaacggtccacatcttcggataaccgcattatcgcggacaggaaccggccggaggtcaatcgccaccgtacaagaaaccggcaggcgattaacctcctttgccgcttccctcggacaaccattttcttttcatcactgcagattagactccgatgaaactttccgtccttctgcattaaagcttgtacgtggagaggctttggttttcaactgcatgctgaatttacctcaccttagttaccgtgcaccggaatcagttgcttcgtttttaaacggagcaaaaacgttgaatccaaagcttgtgacgttggttgaagaagaagttggttctgttattggtgggtttgtggaaaggttcatggactcacttcatcattattcagcggtttttgattcattggaggctggttttccgatgcagaaccgggcccggactttggtggagagggtattttttgggcctagaattgcaggctcgttgggccggatatatagaacgggtggtgaagaggagagaaggtcatggggggagtggttaggtgaggtagggtttaggggagttccggtgagctttgcaaatcattgtcaagcaaagctgttgttagggctttttaatgatgggtatagggttgaagaggtgggtgtgggtagtaataagttggtgttggattggaaatcaagacgtttgctttctgcttccctttggacttgttcttcttcagattctgatttatag
PRIMER_TASK=pick_detection_primers
PRIMER_PICK_RIGHT_PRIMER=1
PRIMER_PICK_INTERNAL_OLIGO=0
PRIMER_PICK_LEFT_PRIMER=1
SEQUENCE_FORCE_RIGHT_START=697
SEQUENCE_FORCE_RIGHT_END=678
PRIMER_NUM_RETURN=3
PRIMER_PICK_ANYWAY=1
PRIMER_WARNING=PRIMER_SALT_DIVALENT > 0.0 but PRIMER_DNTP_CONC <= 0.0; use reasonable value for PRIMER_DNTP_CONC
PRIMER_LEFT_NUM_RETURNED=3
PRIMER_RIGHT_NUM_RETURNED=3
PRIMER_INTERNAL_NUM_RETURNED=0
PRIMER_PAIR_NUM_RETURNED=3
PRIMER_PAIR_0_PENALTY=2.573940
PRIMER_LEFT_0_PENALTY=0.042322
PRIMER_RIGHT_0_PENALTY=2.531618
PRIMER_LEFT_0_SEQUENCE=ctgggggtgcgcataataac
PRIMER_RIGHT_0_SEQUENCE=gcttcgatgatggcttgatt
PRIMER_LEFT_0=526,20
PRIMER_RIGHT_0=697,20
PRIMER_LEFT_0_TM=59.958
PRIMER_RIGHT_0_TM=57.468
PRIMER_LEFT_0_GC_PERCENT=55.000
PRIMER_RIGHT_0_GC_PERCENT=45.000
PRIMER_LEFT_0_SELF_ANY_TH=16.48
PRIMER_RIGHT_0_SELF_ANY_TH=2.34
PRIMER_LEFT_0_SELF_END_TH=0.00
PRIMER_RIGHT_0_SELF_END_TH=0.00
PRIMER_LEFT_0_HAIRPIN_TH=0.00
PRIMER_RIGHT_0_HAIRPIN_TH=0.00
PRIMER_LEFT_0_END_STABILITY=1.8900
PRIMER_RIGHT_0_END_STABILITY=2.5700
PRIMER_PAIR_0_COMPL_ANY_TH=0.00
PRIMER_PAIR_0_COMPL_END_TH=0.00
PRIMER_PAIR_0_PRODUCT_SIZE=172
PRIMER_PAIR_0_PRODUCT_TM=85.8
PRIMER_PAIR_1_PENALTY=2.629283
PRIMER_LEFT_1_PENALTY=0.097666
PRIMER_RIGHT_1_PENALTY=2.531618
PRIMER_LEFT_1_SEQUENCE=ccgaagcccttcatggacta
PRIMER_RIGHT_1_SEQUENCE=gcttcgatgatggcttgatt
PRIMER_LEFT_1=496,20
PRIMER_RIGHT_1=697,20
PRIMER_LEFT_1_TM=60.098
PRIMER_RIGHT_1_TM=57.468
PRIMER_LEFT_1_GC_PERCENT=55.000
PRIMER_RIGHT_1_GC_PERCENT=45.000
PRIMER_LEFT_1_SELF_ANY_TH=9.08
PRIMER_RIGHT_1_SELF_ANY_TH=2.34
PRIMER_LEFT_1_SELF_END_TH=0.00
PRIMER_RIGHT_1_SELF_END_TH=0.00
PRIMER_LEFT_1_HAIRPIN_TH=40.00
PRIMER_RIGHT_1_HAIRPIN_TH=0.00
PRIMER_LEFT_1_END_STABILITY=2.5900
PRIMER_RIGHT_1_END_STABILITY=2.5700
PRIMER_PAIR_1_COMPL_ANY_TH=16.88
PRIMER_PAIR_1_COMPL_END_TH=13.73
PRIMER_PAIR_1_PRODUCT_SIZE=202
PRIMER_PAIR_1_PRODUCT_TM=86.7
PRIMER_PAIR_2_PENALTY=2.631616
PRIMER_LEFT_2_PENALTY=0.099998
PRIMER_RIGHT_2_PENALTY=2.531618
PRIMER_LEFT_2_SEQUENCE=ccgtgacttagctcgagtga
PRIMER_RIGHT_2_SEQUENCE=gcttcgatgatggcttgatt
PRIMER_LEFT_2=401,20
PRIMER_RIGHT_2=697,20
PRIMER_LEFT_2_TM=60.100
PRIMER_RIGHT_2_TM=57.468
PRIMER_LEFT_2_GC_PERCENT=55.000
PRIMER_RIGHT_2_GC_PERCENT=45.000
PRIMER_LEFT_2_SELF_ANY_TH=15.59
PRIMER_RIGHT_2_SELF_ANY_TH=2.34
PRIMER_LEFT_2_SELF_END_TH=0.00
PRIMER_RIGHT_2_SELF_END_TH=0.00
PRIMER_LEFT_2_HAIRPIN_TH=0.00
PRIMER_RIGHT_2_HAIRPIN_TH=0.00
PRIMER_LEFT_2_END_STABILITY=3.4100
PRIMER_RIGHT_2_END_STABILITY=2.5700
PRIMER_PAIR_2_COMPL_ANY_TH=0.00
PRIMER_PAIR_2_COMPL_END_TH=0.00
PRIMER_PAIR_2_PRODUCT_SIZE=297
PRIMER_PAIR_2_PRODUCT_TM=87.6
=
SEQUENCE_ID=Pick start fixed left primer
SEQUENCE_TEMPLATE=atggatttgatggacatggatgccatcaatgacctccacttctctggacacagctccctcaccaacacccctacttccgacgaggattatggttgcacctggaaccactggtctcccatcgtcaactgggacacctttacaggtgccccagatgactttcaccacctcatggacaccatcatcgaggacagaacaactgtcctcgaacagctcagtccatctatcaccacaaccaccaccaccaccacaaccacggatgaggaagaagaagaaatggaaacaacaaccacaacaacaacaacggcaataaaaacgcatgaagttggtgatgattcaaaagggctaaaactagtgcacctattgatggctggcgcggaagccttaaccggttcaaccaaaaaccgtgacttagctcgagtgatattgatccggctcaaggaattagtctcacaacatgctaacggctccaacatggagagacttgccgctcatttcaccgaagcccttcatggactactagaaggtgctgggggtgcgcataataaccaccatcaccacaacaacaacaaacattacctcaccacaaatggtccccacgacaaccaaaacgacacacttgctgctttccaactacttcaagacatgtcaccttacgtcaaattcggccacttcacagccaatcaagccatcatcgaagccgtggcccacgaacgccgcgtccatgtcatcgactacgatatcatggaaggggtccaatgggcctcattaatccaatcacttgcttccaacaacaacggtccacatcttcggataaccgcattatcgcggacaggaaccggccggaggtcaatcgccaccgtacaagaaaccggcaggcgattaacctcctttgccgcttccctcggacaaccattttcttttcatcactgcagattagactccgatgaaactttccgtccttctgcattaaagcttgtacgtggagaggctttggttttcaactgcatgctgaatttacctcaccttagttaccgtgcaccggaatcagttgcttcgtttttaaacggagcaaaaacgttgaatccaaagcttgtgacgttggttgaagaagaagttggttctgttattggtgggtttgtggaaaggttcatggactcacttcatcattattcagcggtttttgattcattggaggctggttttccgatgcagaaccgggcccggactttggtggagagggtattttttgggcctagaattgcaggctcgttgggccggatatatagaacgggtggtgaagaggagagaaggtcatggggggagtggttaggtgaggtagggtttaggggagttccggtgagctttgcaaatcattgtcaagcaaagctgttgttagggctttttaatgatgggtatagggttgaagaggtgggtgtgggtagtaataagttggtgttggattggaaatcaagacgtttgctttctgcttccctttggacttgttcttcttcagattctgatttatag
PRIMER_TASK=pick_detection_primers
PRIMER_PICK_RIGHT_PRIMER=1
PRIMER_PICK_INTERNAL_OLIGO=0
PRIMER_PICK_LEFT_PRIMER=1
SEQUENCE_FORCE_LEFT_START=15
PRIMER_NUM_RETURN=3
PRIMER_PICK_ANYWAY=1
PRIMER_WARNING=PRIMER_SALT_DIVALENT > 0.0 but PRIMER_DNTP_CONC <= 0.0; use reasonable value for PRIMER_DNTP_CONC
PRIMER_LEFT_NUM_RETURNED=3
PRIMER_RIGHT_NUM_RETURNED=3
PRIMER_INTERNAL_NUM_RETURNED=0
PRIMER_PAIR_NUM_RETURNED=3
PRIMER_PAIR_0_PENALTY=2.035489
PRIMER_LEFT_0_PENALTY=1.938159
PRIMER_RIGHT_0_PENALTY=0.097330
PRIMER_LEFT_0_SEQUENCE=atggatgccatcaatgacctc
PRIMER_RIGHT_0_SEQUENCE=aaaggtgtcccagttgacga
PRIMER_LEFT_0=15,21
PRIMER_RIGHT_0=137,20
PRIMER_LEFT_0_TM=59.062
PRIMER_RIGHT_0_TM=60.097
PRIMER_LEFT_0_GC_PERCENT=47.619
PRIMER_RIGHT_0_GC_PERCENT=50.000
PRIMER_LEFT_0_SELF_ANY_TH=7.69
PRIMER_RIGHT_0_SELF_ANY_TH=0.00
PRIMER_LEFT_0_SELF_END_TH=0.00
PRIMER_RIGHT_0_SELF_END_TH=0.00
PRIMER_LEFT_0_HAIRPIN_TH=41.92
PRIMER_RIGHT_0_HAIRPIN_TH=38.67
PRIMER_LEFT_0_END_STABILITY=3.8500
PRIMER_RIGHT_0_END_STABILITY=4.2000
PRIMER_PAIR_0_COMPL_ANY_TH=0.00
PRIMER_PAIR_0_COMPL_END_TH=0.00
PRIMER_PAIR_0_PRODUCT_SIZE=123
PRIMER_PAIR_0_PRODUCT_TM=86.9
PRIMER_PAIR_1_PENALTY=2.036379
PRIMER_LEFT_1_PENALTY=1.938159
PRIMER_RIGHT_1_PENALTY=0.098220
PRIMER_LEFT_1_SEQUENCE=atggatgccatcaatgacctc
PRIMER_RIGHT_1_SEQUENCE=gtcctcgatgatggtgtcca
PRIMER_LEFT_1=15,21
PRIMER_RIGHT_1=188,20
PRIMER_LEFT_1_TM=59.062
PRIMER_RIGHT_1_TM=60.098
PRIMER_LEFT_1_GC_PERCENT=47.619
PRIMER_RIGHT_1_GC_PERCENT=55.000
PRIMER_LEFT_1_SELF_ANY_TH=7.69
PRIMER_RIGHT_1_SELF_ANY_TH=0.00
PRIMER_LEFT_1_SELF_END_TH=0.00
PRIMER_RIGHT_1_SELF_END_TH=0.00
PRIMER_LEFT_1_HAIRPIN_TH=41.92
PRIMER_RIGHT_1_HAIRPIN_TH=38.57
PRIMER_LEFT_1_END_STABILITY=3.8500
PRIMER_RIGHT_1_END_STABILITY=4.0200
PRIMER_PAIR_1_COMPL_ANY_TH=20.44
PRIMER_PAIR_1_COMPL_END_TH=20.44
PRIMER_PAIR_1_PRODUCT_SIZE=174
PRIMER_PAIR_1_PRODUCT_TM=88.5
PRIMER_PAIR_2_PENALTY=2.036379
PRIMER_LEFT_2_PENALTY=1.938159
PRIMER_RIGHT_2_PENALTY=0.098220
PRIMER_LEFT_2_SEQUENCE=atggatgccatcaatgacctc
PRIMER_RIGHT_2_SEQUENCE=tgtcctcgatgatggtgtcc
PRIMER_LEFT_2=15,21
PRIMER_RIGHT_2=189,20
PRIMER_LEFT_2_TM=59.062
PRIMER_RIGHT_2_TM=60.098
PRIMER_LEFT_2_GC_PERCENT=47.619
PRIMER_RIGHT_2_GC_PERCENT=55.000
PRIMER_LEFT_2_SELF_ANY_TH=7.69
PRIMER_RIGHT_2_SELF_ANY_TH=0.00
PRIMER_LEFT_2_SELF_END_TH=0.00
PRIMER_RIGHT_2_SELF_END_TH=0.00
PRIMER_LEFT_2_HAIRPIN_TH=41.92
PRIMER_RIGHT_2_HAIRPIN_TH=0.00
PRIMER_LEFT_2_END_STABILITY=3.8500
PRIMER_RIGHT_2_END_STABILITY=4.0200
PRIMER_PAIR_2_COMPL_ANY_TH=15.21
PRIMER_PAIR_2_COMPL_END_TH=15.21
PRIMER_PAIR_2_PRODUCT_SIZE=175
PRIMER_PAIR_2_PRODUCT_TM=88.4
=
SEQUENCE_ID=Pick start fixed right primer
SEQUENCE_TEMPLATE=atggatttgatggacatggatgccatcaatgacctccacttctctggacacagctccctcaccaacacccctacttccgacgaggattatggttgcacctggaaccactggtctcccatcgtcaactgggacacctttacaggtgccccagatgactttcaccacctcatggacaccatcatcgaggacagaacaactgtcctcgaacagctcagtccatctatcaccacaaccaccaccaccaccacaaccacggatgaggaagaagaagaaatggaaacaacaaccacaacaacaacaacggcaataaaaacgcatgaagttggtgatgattcaaaagggctaaaactagtgcacctattgatggctggcgcggaagccttaaccggttcaaccaaaaaccgtgacttagctcgagtgatattgatccggctcaaggaattagtctcacaacatgctaacggctccaacatggagagacttgccgctcatttcaccgaagcccttcatggactactagaaggtgctgggggtgcgcataataaccaccatcaccacaacaacaacaaacattacctcaccacaaatggtccccacgacaaccaaaacgacacacttgctgctttccaactacttcaagacatgtcaccttacgtcaaattcggccacttcacagccaatcaagccatcatcgaagccgtggcccacgaacgccgcgtccatgtcatcgactacgatatcatggaaggggtccaatgggcctcattaatccaatcacttgcttccaacaacaacggtccacatcttcggataaccgcattatcgcggacaggaaccggccggaggtcaatcgccaccgtacaagaaaccggcaggcgattaacctcctttgccgcttccctcggacaaccattttcttttcatcactgcagattagactccgatgaaactttccgtccttctgcattaaagcttgtacgtggagaggctttggttttcaactgcatgctgaatttacctcaccttagttaccgtgcaccggaatcagttgcttcgtttttaaacggagcaaaaacgttgaatccaaagcttgtgacgttggttgaagaagaagttggttctgttattggtgggtttgtggaaaggttcatggactcacttcatcattattcagcggtttttgattcattggaggctggttttccgatgcagaaccgggcccggactttggtggagagggtattttttgggcctagaattgcaggctcgttgggccggatatatagaacgggtggtgaagaggagagaaggtcatggggggagtggttaggtgaggtagggtttaggggagttccggtgagctttgcaaatcattgtcaagcaaagctgttgttagggctttttaatgatgggtatagggttgaagaggtgggtgtgggtagtaataagttggtgttggattggaaatcaagacgtttgctttctgcttccctttggacttgttcttcttcagattctgatttatag
PRIMER_TASK=pick_detection_primers
PRIMER_PICK_RIGHT_PRIMER=1
PRIMER_PICK_INTERNAL_OLIGO=0
PRIMER_PICK_LEFT_PRIMER=1
SEQUENCE_FORCE_RIGHT_START=697
PRIMER_NUM_RETURN=3
PRIMER_PICK_ANYWAY=1
PRIMER_WARNING=PRIMER_SALT_DIVALENT > 0.0 but PRIMER_DNTP_CONC <= 0.0; use reasonable value for PRIMER_DNTP_CONC
PRIMER_LEFT_NUM_RETURNED=3
PRIMER_RIGHT_NUM_RETURNED=3
PRIMER_INTERNAL_NUM_RETURNED=0
PRIMER_PAIR_NUM_RETURNED=3
PRIMER_PAIR_0_PENALTY=2.139074
PRIMER_LEFT_0_PENALTY=0.042322
PRIMER_RIGHT_0_PENALTY=2.096751
PRIMER_LEFT_0_SEQUENCE=ctgggggtgcgcataataac
PRIMER_RIGHT_0_SEQUENCE=gcttcgatgatggcttgattg
PRIMER_LEFT_0=526,20
PRIMER_RIGHT_0=697,21
PRIMER_LEFT_0_TM=59.958
PRIMER_RIGHT_0_TM=58.903
PRIMER_LEFT_0_GC_PERCENT=55.000
PRIMER_RIGHT_0_GC_PERCENT=47.619
PRIMER_LEFT_0_SELF_ANY_TH=16.48
PRIMER_RIGHT_0_SELF_ANY_TH=2.34
PRIMER_LEFT_0_SELF_END_TH=0.00
PRIMER_RIGHT_0_SELF_END_TH=0.00
PRIMER_LEFT_0_HAIRPIN_TH=0.00
PRIMER_RIGHT_0_HAIRPIN_TH=0.00
PRIMER_LEFT_0_END_STABILITY=1.8900
PRIMER_RIGHT_0_END_STABILITY=2.6700
PRIMER_PAIR_0_COMPL_ANY_TH=0.00
PRIMER_PAIR_0_COMPL_END_TH=0.00
PRIMER_PAIR_0_PRODUCT_SIZE=172
PRIMER_PAIR_0_PRODUCT_TM=85.8
PRIMER_PAIR_1_PENALTY=2.194417
PRIMER_LEFT_1_PENALTY=0.097666
PRIMER_RIGHT_1_PENALTY=2.096751
PRIMER_LEFT_1_SEQUENCE=ccgaagcccttcatggacta
PRIMER_RIGHT_1_SEQUENCE=gcttcgatgatggcttgattg
PRIMER_LEFT_1=496,20
PRIMER_RIGHT_1=697,21
PRIMER_LEFT_1_TM=60.098
PRIMER_RIGHT_1_TM=58.903
PRIMER_LEFT_1_GC_PERCENT=55.000
PRIMER_RIGHT_1_GC_PERCENT=47.619
PRIMER_LEFT_1_SELF_ANY_TH=9.08
PRIMER_RIGHT_1_SELF_ANY_TH=2.34
PRIMER_LEFT_1_SELF_END_TH=0.00
PRIMER_RIGHT_1_SELF_END_TH=0.00
PRIMER_LEFT_1_HAIRPIN_TH=40.00
PRIMER_RIGHT_1_HAIRPIN_TH=0.00
PRIMER_LEFT_1_END_STABILITY=2.5900
PRIMER_RIGHT_1_END_STABILITY=2.6700
PRIMER_PAIR_1_COMPL_ANY_TH=16.88
PRIMER_PAIR_1_COMPL_END_TH=13.73
PRIMER_PAIR_1_PRODUCT_SIZE=202
PRIMER_PAIR_1_PRODUCT_TM=86.7
PRIMER_PAIR_2_PENALTY=2.196749
PRIMER_LEFT_2_PENALTY=0.099998
PRIMER_RIGHT_2_PENALTY=2.096751
PRIMER_LEFT_2_SEQUENCE=ccgtgacttagctcgagtga
PRIMER_RIGHT_2_SEQUENCE=gcttcgatgatggcttgattg
PRIMER_LEFT_2=401,20
PRIMER_RIGHT_2=697,21
PRIMER_LEFT_2_TM=60.100
PRIMER_RIGHT_2_TM=58.903
PRIMER_LEFT_2_GC_PERCENT=55.000
PRIMER_RIGHT_2_GC_PERCENT=47.619
PRIMER_LEFT_2_SELF_ANY_TH=15.59
PRIMER_RIGHT_2_SELF_ANY_TH=2.34
PRIMER_LEFT_2_SELF_END_TH=0.00
PRIMER_RIGHT_2_SELF_END_TH=0.00
PRIMER_LEFT_2_HAIRPIN_TH=0.00
PRIMER_RIGHT_2_HAIRPIN_TH=0.00
PRIMER_LEFT_2_END_STABILITY=3.4100
PRIMER_RIGHT_2_END_STABILITY=2.6700
PRIMER_PAIR_2_COMPL_ANY_TH=0.00
PRIMER_PAIR_2_COMPL_END_TH=0.00
PRIMER_PAIR_2_PRODUCT_SIZE=297
PRIMER_PAIR_2_PRODUCT_TM=87.6
=
SEQUENCE_ID=Pick end fixed left primer
SEQUENCE_TEMPLATE=atggatttgatggacatggatgccatcaatgacctccacttctctggacacagctccctcaccaacacccctacttccgacgaggattatggttgcacctggaaccactggtctcccatcgtcaactgggacacctttacaggtgccccagatgactttcaccacctcatggacaccatcatcgaggacagaacaactgtcctcgaacagctcagtccatctatcaccacaaccaccaccaccaccacaaccacggatgaggaagaagaagaaatggaaacaacaaccacaacaacaacaacggcaataaaaacgcatgaagttggtgatgattcaaaagggctaaaactagtgcacctattgatggctggcgcggaagccttaaccggttcaaccaaaaaccgtgacttagctcgagtgatattgatccggctcaaggaattagtctcacaacatgctaacggctccaacatggagagacttgccgctcatttcaccgaagcccttcatggactactagaaggtgctgggggtgcgcataataaccaccatcaccacaacaacaacaaacattacctcaccacaaatggtccccacgacaaccaaaacgacacacttgctgctttccaactacttcaagacatgtcaccttacgtcaaattcggccacttcacagccaatcaagccatcatcgaagccgtggcccacgaacgccgcgtccatgtcatcgactacgatatcatggaaggggtccaatgggcctcattaatccaatcacttgcttccaacaacaacggtccacatcttcggataaccgcattatcgcggacaggaaccggccggaggtcaatcgccaccgtacaagaaaccggcaggcgattaacctcctttgccgcttccctcggacaaccattttcttttcatcactgcagattagactccgatgaaactttccgtccttctgcattaaagcttgtacgtggagaggctttggttttcaactgcatgctgaatttacctcaccttagttaccgtgcaccggaatcagttgcttcgtttttaaacggagcaaaaacgttgaatccaaagcttgtgacgttggttgaagaagaagttggttctgttattggtgggtttgtggaaaggttcatggactcacttcatcattattcagcggtttttgattcattggaggctggttttccgatgcagaaccgggcccggactttggtggagagggtattttttgggcctagaattgcaggctcgttgggccggatatatagaacgggtggtgaagaggagagaaggtcatggggggagtggttaggtgaggtagggtttaggggagttccggtgagctttgcaaatcattgtcaagcaaagctgttgttagggctttttaatgatgggtatagggttgaagaggtgggtgtgggtagtaataagttggtgttggattggaaatcaagacgtttgctttctgcttccctttggacttgttcttcttcagattctgatttatag
PRIMER_TASK=pick_detection_primers
PRIMER_PICK_RIGHT_PRIMER=1
PRIMER_PICK_INTERNAL_OLIGO=0
PRIMER_PICK_LEFT_PRIMER=1
SEQUENCE_FORCE_LEFT_END=34
PRIMER_NUM_RETURN=2
PRIMER_PICK_ANYWAY=1
PRIMER_WARNING=PRIMER_SALT_DIVALENT > 0.0 but PRIMER_DNTP_CONC <= 0.0; use reasonable value for PRIMER_DNTP_CONC
PRIMER_LEFT_NUM_RETURNED=2
PRIMER_RIGHT_NUM_RETURNED=2
PRIMER_INTERNAL_NUM_RETURNED=0
PRIMER_PAIR_NUM_RETURNED=2
PRIMER_PAIR_0_PENALTY=1.757074
PRIMER_LEFT_0_PENALTY=1.659744
PRIMER_RIGHT_0_PENALTY=0.097330
PRIMER_LEFT_0_SEQUENCE=catggatgccatcaatgacct
PRIMER_RIGHT_0_SEQUENCE=aaaggtgtcccagttgacga
PRIMER_LEFT_0=14,21
PRIMER_RIGHT_0=137,20
PRIMER_LEFT_0_TM=59.340
PRIMER_RIGHT_0_TM=60.097
PRIMER_LEFT_0_GC_PERCENT=47.619
PRIMER_RIGHT_0_GC_PERCENT=50.000
PRIMER_LEFT_0_SELF_ANY_TH=16.82
PRIMER_RIGHT_0_SELF_ANY_TH=0.00
PRIMER_LEFT_0_SELF_END_TH=0.00
PRIMER_RIGHT_0_SELF_END_TH=0.00
PRIMER_LEFT_0_HAIRPIN_TH=44.12
PRIMER_RIGHT_0_HAIRPIN_TH=38.67
PRIMER_LEFT_0_END_STABILITY=3.8500
PRIMER_RIGHT_0_END_STABILITY=4.2000
PRIMER_PAIR_0_COMPL_ANY_TH=0.00
PRIMER_PAIR_0_COMPL_END_TH=0.00
PRIMER_PAIR_0_PRODUCT_SIZE=124
PRIMER_PAIR_0_PRODUCT_TM=87.1
PRIMER_PAIR_1_PENALTY=1.757964
PRIMER_LEFT_1_PENALTY=1.659744
PRIMER_RIGHT_1_PENALTY=0.098220
PRIMER_LEFT_1_SEQUENCE=catggatgccatcaatgacct
PRIMER_RIGHT_1_SEQUENCE=gtcctcgatgatggtgtcca
PRIMER_LEFT_1=14,21
PRIMER_RIGHT_1=188,20
PRIMER_LEFT_1_TM=59.340
PRIMER_RIGHT_1_TM=60.098
PRIMER_LEFT_1_GC_PERCENT=47.619
PRIMER_RIGHT_1_GC_PERCENT=55.000
PRIMER_LEFT_1_SELF_ANY_TH=16.82
PRIMER_RIGHT_1_SELF_ANY_TH=0.00
PRIMER_LEFT_1_SELF_END_TH=0.00
PRIMER_RIGHT_1_SELF_END_TH=0.00
PRIMER_LEFT_1_HAIRPIN_TH=44.12
PRIMER_RIGHT_1_HAIRPIN_TH=38.57
PRIMER_LEFT_1_END_STABILITY=3.8500
PRIMER_RIGHT_1_END_STABILITY=4.0200
PRIMER_PAIR_1_COMPL_ANY_TH=20.44
PRIMER_PAIR_1_COMPL_END_TH=20.44
PRIMER_PAIR_1_PRODUCT_SIZE=175
PRIMER_PAIR_1_PRODUCT_TM=88.6
=
SEQUENCE_ID=Pick end fixed right primer
SEQUENCE_TEMPLATE=atggatttgatggacatggatgccatcaatgacctccacttctctggacacagctccctcaccaacacccctacttccgacgaggattatggttgcacctggaaccactggtctcccatcgtcaactgggacacctttacaggtgccccagatgactttcaccacctcatggacaccatcatcgaggacagaacaactgtcctcgaacagctcagtccatctatcaccacaaccaccaccaccaccacaaccacggatgaggaagaagaagaaatggaaacaacaaccacaacaacaacaacggcaataaaaacgcatgaagttggtgatgattcaaaagggctaaaactagtgcacctattgatggctggcgcggaagccttaaccggttcaaccaaaaaccgtgacttagctcgagtgatattgatccggctcaaggaattagtctcacaacatgctaacggctccaacatggagagacttgccgctcatttcaccgaagcccttcatggactactagaaggtgctgggggtgcgcataataaccaccatcaccacaacaacaacaaacattacctcaccacaaatggtccccacgacaaccaaaacgacacacttgctgctttccaactacttcaagacatgtcaccttacgtcaaattcggccacttcacagccaatcaagccatcatcgaagccgtggcccacgaacgccgcgtccatgtcatcgactacgatatcatggaaggggtccaatgggcctcattaatccaatcacttgcttccaacaacaacggtccacatcttcggataaccgcattatcgcggacaggaaccggccggaggtcaatcgccaccgtacaagaaaccggcaggcgattaacctcctttgccgcttccctcggacaaccattttcttttcatcactgcagattagactccgatgaaactttccgtccttctgcattaaagcttgtacgtggagaggctttggttttcaactgcatgctgaatttacctcaccttagttaccgtgcaccggaatcagttgcttcgtttttaaacggagcaaaaacgttgaatccaaagcttgtgacgttggttgaagaagaagttggttctgttattggtgggtttgtggaaaggttcatggactcacttcatcattattcagcggtttttgattcattggaggctggttttccgatgcagaaccgggcccggactttggtggagagggtattttttgggcctagaattgcaggctcgttgggccggatatatagaacgggtggtgaagaggagagaaggtcatggggggagtggttaggtgaggtagggtttaggggagttccggtgagctttgcaaatcattgtcaagcaaagctgttgttagggctttttaatgatgggtatagggttgaagaggtgggtgtgggtagtaataagttggtgttggattggaaatcaagacgtttgctttctgcttccctttggacttgttcttcttcagattctgatttatag
PRIMER_TASK=pick_detection_primers
PRIMER_PICK_RIGHT_PRIMER=1
PRIMER_PICK_INTERNAL_OLIGO=0
PRIMER_PICK_LEFT_PRIMER=1
SEQUENCE_FORCE_RIGHT_END=678
PRIMER_NUM_RETURN=2
PRIMER_PICK_ANYWAY=1
PRIMER_WARNING=PRIMER_SALT_DIVALENT > 0.0 but PRIMER_DNTP_CONC <= 0.0; use reasonable value for PRIMER_DNTP_CONC
PRIMER_LEFT_NUM_RETURNED=2
PRIMER_RIGHT_NUM_RETURNED=2
PRIMER_INTERNAL_NUM_RETURNED=0
PRIMER_PAIR_NUM_RETURNED=2
PRIMER_PAIR_0_PENALTY=1.074493
PRIMER_LEFT_0_PENALTY=0.042322
PRIMER_RIGHT_0_PENALTY=1.032171
PRIMER_LEFT_0_SEQUENCE=ctgggggtgcgcataataac
PRIMER_RIGHT_0_SEQUENCE=ggcttcgatgatggcttgatt
PRIMER_LEFT_0=526,20
PRIMER_RIGHT_0=698,21
PRIMER_LEFT_0_TM=59.958
PRIMER_RIGHT_0_TM=59.968
PRIMER_LEFT_0_GC_PERCENT=55.000
PRIMER_RIGHT_0_GC_PERCENT=47.619
PRIMER_LEFT_0_SELF_ANY_TH=16.48
PRIMER_RIGHT_0_SELF_ANY_TH=2.34
PRIMER_LEFT_0_SELF_END_TH=0.00
PRIMER_RIGHT_0_SELF_END_TH=0.00
PRIMER_LEFT_0_HAIRPIN_TH=0.00
PRIMER_RIGHT_0_HAIRPIN_TH=31.99
PRIMER_LEFT_0_END_STABILITY=1.8900
PRIMER_RIGHT_0_END_STABILITY=2.5700
PRIMER_PAIR_0_COMPL_ANY_TH=0.00
PRIMER_PAIR_0_COMPL_END_TH=0.00
PRIMER_PAIR_0_PRODUCT_SIZE=173
PRIMER_PAIR_0_PRODUCT_TM=86.0
PRIMER_PAIR_1_PENALTY=1.129837
PRIMER_LEFT_1_PENALTY=0.097666
PRIMER_RIGHT_1_PENALTY=1.032171
PRIMER_LEFT_1_SEQUENCE=ccgaagcccttcatggacta
PRIMER_RIGHT_1_SEQUENCE=ggcttcgatgatggcttgatt
PRIMER_LEFT_1=496,20
PRIMER_RIGHT_1=698,21
PRIMER_LEFT_1_TM=60.098
PRIMER_RIGHT_1_TM=59.968
PRIMER_LEFT_1_GC_PERCENT=55.000
PRIMER_RIGHT_1_GC_PERCENT=47.619
PRIMER_LEFT_1_SELF_ANY_TH=9.08
PRIMER_RIGHT_1_SELF_ANY_TH=2.34
PRIMER_LEFT_1_SELF_END_TH=0.00
PRIMER_RIGHT_1_SELF_END_TH=0.00
PRIMER_LEFT_1_HAIRPIN_TH=40.00
PRIMER_RIGHT_1_HAIRPIN_TH=31.99
PRIMER_LEFT_1_END_STABILITY=2.5900
PRIMER_RIGHT_1_END_STABILITY=2.5700
PRIMER_PAIR_1_COMPL_ANY_TH=23.58
PRIMER_PAIR_1_COMPL_END_TH=23.58
PRIMER_PAIR_1_PRODUCT_SIZE=203
PRIMER_PAIR_1_PRODUCT_TM=86.8
=
SEQUENCE_ID=Pick cloning primers
SEQUENCE_TEMPLATE=atggatttgatggacatggatgccatcaatgacctccacttctctggacacagctccctcaccaacacccctacttccgacgaggattatggttgcacctggaaccactggtctcccatcgtcaactgggacacctttacaggtgccccagatgactttcaccacctcatggacaccatcatcgaggacagaacaactgtcctcgaacagctcagtccatctatcaccacaaccaccaccaccaccacaaccacggatgaggaagaagaagaaatggaaacaacaaccacaacaacaacaacggcaataaaaacgcatgaagttggtgatgattcaaaagggctaaaactagtgcacctattgatggctggcgcggaagccttaaccggttcaaccaaaaaccgtgacttagctcgagtgatattgatccggctcaaggaattagtctcacaacatgctaacggctccaacatggagagacttgccgctcatttcaccgaagcccttcatggactactagaaggtgctgggggtgcgcataataaccaccatcaccacaacaacaacaaacattacctcaccacaaatggtccccacgacaaccaaaacgacacacttgctgctttccaactacttcaagacatgtcaccttacgtcaaattcggccacttcacagccaatcaagccatcatcgaagccgtggcccacgaacgccgcgtccatgtcatcgactacgatatcatggaaggggtccaatgggcctcattaatccaatcacttgcttccaacaacaacggtccacatcttcggataaccgcattatcgcggacaggaaccggccggaggtcaatcgccaccgtacaagaaaccggcaggcgattaacctcctttgccgcttccctcggacaaccattttcttttcatcactgcagattagactccgatgaaactttccgtccttctgcattaaagcttgtacgtggagaggctttggttttcaactgcatgctgaatttacctcaccttagttaccgtgcaccggaatcagttgcttcgtttttaaacggagcaaaaacgttgaatccaaagcttgtgacgttggttgaagaagaagttggttctgttattggtgggtttgtggaaaggttcatggactcacttcatcattattcagcggtttttgattcattggaggctggttttccgatgcagaaccgggcccggactttggtggagagggtattttttgggcctagaattgcaggctcgttgggccggatatatagaacgggtggtgaagaggagagaaggtcatggggggagtggttaggtgaggtagggtttaggggagttccggtgagctttgcaaatcattgtcaagcaaagctgttgttagggctttttaatgatgggtatagggttgaagaggtgggtgtgggtagtaataagttggtgttggattggaaatcaagacgtttgctttctgcttccctttggacttgttcttcttcagattctgatttatag
PRIMER_TASK=pick_cloning_primers
PRIMER_PICK_RIGHT_PRIMER=1
PRIMER_PICK_INTERNAL_OLIGO=0
PRIMER_PICK_LEFT_PRIMER=1
SEQUENCE_INCLUDED_REGION=15,203
PRIMER_NUM_RETURN=2
PRIMER_PICK_ANYWAY=1
PRIMER_WARNING=PRIMER_SALT_DIVALENT > 0.0 but PRIMER_DNTP_CONC <= 0.0; use reasonable value for PRIMER_DNTP_CONC
PRIMER_LEFT_NUM_RETURNED=2
PRIMER_RIGHT_NUM_RETURNED=2
PRIMER_INTERNAL_NUM_RETURNED=0
PRIMER_PAIR_NUM_RETURNED=2
PRIMER_PAIR_0_PENALTY=2.951836
PRIMER_LEFT_0_PENALTY=1.938159
PRIMER_RIGHT_0_PENALTY=1.013677
PRIMER_LEFT_0_SEQUENCE=atggatgccatcaatgacctc
PRIMER_RIGHT_0_SEQUENCE=ggactgagctgttcgaggac
PRIMER_LEFT_0=15,21
PRIMER_RIGHT_0=217,20
PRIMER_LEFT_0_TM=59.062
PRIMER_RIGHT_0_TM=61.014
PRIMER_LEFT_0_GC_PERCENT=47.619
PRIMER_RIGHT_0_GC_PERCENT=60.000
PRIMER_LEFT_0_SELF_ANY_TH=7.69
PRIMER_RIGHT_0_SELF_ANY_TH=0.00
PRIMER_LEFT_0_SELF_END_TH=0.00
PRIMER_RIGHT_0_SELF_END_TH=0.00
PRIMER_LEFT_0_HAIRPIN_TH=41.92
PRIMER_RIGHT_0_HAIRPIN_TH=0.00
PRIMER_LEFT_0_END_STABILITY=3.8500
PRIMER_RIGHT_0_END_STABILITY=3.8500
PRIMER_PAIR_0_COMPL_ANY_TH=0.00
PRIMER_PAIR_0_COMPL_END_TH=0.00
PRIMER_PAIR_0_PRODUCT_SIZE=203
PRIMER_PAIR_0_PRODUCT_TM=88.8
PRIMER_PAIR_1_PENALTY=3.136630
PRIMER_LEFT_1_PENALTY=2.122953
PRIMER_RIGHT_1_PENALTY=1.013677
PRIMER_LEFT_1_SEQUENCE=atggatgccatcaatgacct
PRIMER_RIGHT_1_SEQUENCE=ggactgagctgttcgaggac
PRIMER_LEFT_1=15,20
PRIMER_RIGHT_1=217,20
PRIMER_LEFT_1_TM=57.877
PRIMER_RIGHT_1_TM=61.014
PRIMER_LEFT_1_GC_PERCENT=45.000
PRIMER_RIGHT_1_GC_PERCENT=60.000
PRIMER_LEFT_1_SELF_ANY_TH=7.69
PRIMER_RIGHT_1_SELF_ANY_TH=0.00
PRIMER_LEFT_1_SELF_END_TH=0.00
PRIMER_RIGHT_1_SELF_END_TH=0.00
PRIMER_LEFT_1_HAIRPIN_TH=41.92
PRIMER_RIGHT_1_HAIRPIN_TH=0.00
PRIMER_LEFT_1_END_STABILITY=3.8500
PRIMER_RIGHT_1_END_STABILITY=3.8500
PRIMER_PAIR_1_COMPL_ANY_TH=0.00
PRIMER_PAIR_1_COMPL_END_TH=0.00
PRIMER_PAIR_1_PRODUCT_SIZE=203
PRIMER_PAIR_1_PRODUCT_TM=88.8
=
SEQUENCE_ID=Pick discriminative primers
SEQUENCE_TEMPLATE=atggatttgatggacatggatgccatcaatgacctccacttctctggacacagctccctcaccaacacccctacttccgacgaggattatggttgcacctggaaccactggtctcccatcgtcaactgggacacctttacaggtgccccagatgactttcaccacctcatggacaccatcatcgaggacagaacaactgtcctcgaacagctcagtccatctatcaccacaaccaccaccaccaccacaaccacggatgaggaagaagaagaaatggaaacaacaaccacaacaacaacaacggcaataaaaacgcatgaagttggtgatgattcaaaagggctaaaactagtgcacctattgatggctggcgcggaagccttaaccggttcaaccaaaaaccgtgacttagctcgagtgatattgatccggctcaaggaattagtctcacaacatgctaacggctccaacatggagagacttgccgctcatttcaccgaagcccttcatggactactagaaggtgctgggggtgcgcataataaccaccatcaccacaacaacaacaaacattacctcaccacaaatggtccccacgacaaccaaaacgacacacttgctgctttccaactacttcaagacatgtcaccttacgtcaaattcggccacttcacagccaatcaagccatcatcgaagccgtggcccacgaacgccgcgtccatgtcatcgactacgatatcatggaaggggtccaatgggcctcattaatccaatcacttgcttccaacaacaacggtccacatcttcggataaccgcattatcgcggacaggaaccggccggaggtcaatcgccaccgtacaagaaaccggcaggcgattaacctcctttgccgcttccctcggacaaccattttcttttcatcactgcagattagactccgatgaaactttccgtccttctgcattaaagcttgtacgtggagaggctttggttttcaactgcatgctgaatttacctcaccttagttaccgtgcaccggaatcagttgcttcgtttttaaacggagcaaaaacgttgaatccaaagcttgtgacgttggttgaagaagaagttggttctgttattggtgggtttgtggaaaggttcatggactcacttcatcattattcagcggtttttgattcattggaggctggttttccgatgcagaaccgggcccggactttggtggagagggtattttttgggcctagaattgcaggctcgttgggccggatatatagaacgggtggtgaagaggagagaaggtcatggggggagtggttaggtgaggtagggtttaggggagttccggtgagctttgcaaatcattgtcaagcaaagctgttgttagggctttttaatgatgggtatagggttgaagaggtgggtgtgggtagtaataagttggtgttggattggaaatcaagacgtttgctttctgcttccctttggacttgttcttcttcagattctgatttatag
PRIMER_TASK=pick_discriminative_primers
PRIMER_PICK_RIGHT_PRIMER=1
PRIMER_PICK_INTERNAL_OLIGO=0
PRIMER_PICK_LEFT_PRIMER=1
SEQUENCE_TARGET=35,163
PRIMER_NUM_RETURN=2
PRIMER_PICK_ANYWAY=1
PRIMER_WARNING=PRIMER_SALT_DIVALENT > 0.0 but PRIMER_DNTP_CONC <= 0.0; use reasonable value for PRIMER_DNTP_CONC
PRIMER_LEFT_NUM_RETURNED=2
PRIMER_RIGHT_NUM_RETURNED=2
PRIMER_INTERNAL_NUM_RETURNED=0
PRIMER_PAIR_NUM_RETURNED=2
PRIMER_PAIR_0_PENALTY=2.673422
PRIMER_LEFT_0_PENALTY=1.659744
PRIMER_RIGHT_0_PENALTY=1.013677
PRIMER_LEFT_0_SEQUENCE=catggatgccatcaatgacct
PRIMER_RIGHT_0_SEQUENCE=ggactgagctgttcgaggac
PRIMER_LEFT_0=14,21
PRIMER_RIGHT_0=217,20
PRIMER_LEFT_0_TM=59.340
PRIMER_RIGHT_0_TM=61.014
PRIMER_LEFT_0_GC_PERCENT=47.619
PRIMER_RIGHT_0_GC_PERCENT=60.000
PRIMER_LEFT_0_SELF_ANY_TH=16.82
PRIMER_RIGHT_0_SELF_ANY_TH=0.00
PRIMER_LEFT_0_SELF_END_TH=0.00
PRIMER_RIGHT_0_SELF_END_TH=0.00
PRIMER_LEFT_0_HAIRPIN_TH=44.12
PRIMER_RIGHT_0_HAIRPIN_TH=0.00
PRIMER_LEFT_0_END_STABILITY=3.8500
PRIMER_RIGHT_0_END_STABILITY=3.8500
PRIMER_PAIR_0_COMPL_ANY_TH=0.00
PRIMER_PAIR_0_COMPL_END_TH=0.00
PRIMER_PAIR_0_PRODUCT_SIZE=204
PRIMER_PAIR_0_PRODUCT_TM=89.0
PRIMER_PAIR_1_PENALTY=3.136630
PRIMER_LEFT_1_PENALTY=2.122953
PRIMER_RIGHT_1_PENALTY=1.013677
PRIMER_LEFT_1_SEQUENCE=atggatgccatcaatgacct
PRIMER_RIGHT_1_SEQUENCE=ggactgagctgttcgaggac
PRIMER_LEFT_1=15,20
PRIMER_RIGHT_1=217,20
PRIMER_LEFT_1_TM=57.877
PRIMER_RIGHT_1_TM=61.014
PRIMER_LEFT_1_GC_PERCENT=45.000
PRIMER_RIGHT_1_GC_PERCENT=60.000
PRIMER_LEFT_1_SELF_ANY_TH=7.69
PRIMER_RIGHT_1_SELF_ANY_TH=0.00
PRIMER_LEFT_1_SELF_END_TH=0.00
PRIMER_RIGHT_1_SELF_END_TH=0.00
PRIMER_LEFT_1_HAIRPIN_TH=41.92
PRIMER_RIGHT_1_HAIRPIN_TH=0.00
PRIMER_LEFT_1_END_STABILITY=3.8500
PRIMER_RIGHT_1_END_STABILITY=3.8500
PRIMER_PAIR_1_COMPL_ANY_TH=0.00
PRIMER_PAIR_1_COMPL_END_TH=0.00
PRIMER_PAIR_1_PRODUCT_SIZE=203
PRIMER_PAIR_1_PRODUCT_TM=88.8
=
SEQUENCE_ID=Pick start fixed left primer unique
SEQUENCE_TEMPLATE=atggatttgatggacatggatgccatcaatgacctccacttctctggacacagctccctcaccaacacccctacttccgacgaggattatggttgcacctggaaccactggtctcccatcgtcaactgggacacctttacaggtgccccagatgactttcaccacctcatggacaccatcatcgaggacagaacaactgtcctcgaacagctcagtccatctatcaccacaaccaccaccaccaccacaaccacggatgaggaagaagaagaaatggaaacaacaaccacaacaacaacaacggcaataaaaacgcatgaagttggtgatgattcaaaagggctaaaactagtgcacctattgatggctggcgcggaagccttaaccggttcaaccaaaaaccgtgacttagctcgagtgatattgatccggctcaaggaattagtctcacaacatgctaacggctccaacatggagagacttgccgctcatttcaccgaagcccttcatggactactagaaggtgctgggggtgcgcataataaccaccatcaccacaacaacaacaaacattacctcaccacaaatggtccccacgacaaccaaaacgacacacttgctgctttccaactacttcaagacatgtcaccttacgtcaaattcggccacttcacagccaatcaagccatcatcgaagccgtggcccacgaacgccgcgtccatgtcatcgactacgatatcatggaaggggtccaatgggcctcattaatccaatcacttgcttccaacaacaacggtccacatcttcggataaccgcattatcgcggacaggaaccggccggaggtcaatcgccaccgtacaagaaaccggcaggcgattaacctcctttgccgcttccctcggacaaccattttcttttcatcactgcagattagactccgatgaaactttccgtccttctgcattaaagcttgtacgtggagaggctttggttttcaactgcatgctgaatttacctcaccttagttaccgtgcaccggaatcagttgcttcgtttttaaacggagcaaaaacgttgaatccaaagcttgtgacgttggttgaagaagaagttggttctgttattggtgggtttgtggaaaggttcatggactcacttcatcattattcagcggtttttgattcattggaggctggttttccgatgcagaaccgggcccggactttggtggagagggtattttttgggcctagaattgcaggctcgttgggccggatatatagaacgggtggtgaagaggagagaaggtcatggggggagtggttaggtgaggtagggtttaggggagttccggtgagctttgcaaatcattgtcaagcaaagctgttgttagggctttttaatgatgggtatagggttgaagaggtgggtgtgggtagtaataagttggtgttggattggaaatcaagacgtttgctttctgcttccctttggacttgttcttcttcagattctgatttatag
PRIMER_TASK=pick_detection_primers
PRIMER_PICK_RIGHT_PRIMER=1
PRIMER_PICK_INTERNAL_OLIGO=0
PRIMER_PICK_LEFT_PRIMER=1
SEQUENCE_FORCE_LEFT_START=15
PRIMER_NUM_RETURN=3
PRIMER_MIN_THREE_PRIME_DISTANCE=0
PRIMER_PICK_ANYWAY=1
PRIMER_WARNING=PRIMER_SALT_DIVALENT > 0.0 but PRIMER_DNTP_CONC <= 0.0; use reasonable value for PRIMER_DNTP_CONC
PRIMER_LEFT_NUM_RETURNED=3
PRIMER_RIGHT_NUM_RETURNED=3
PRIMER_INTERNAL_NUM_RETURNED=0
PRIMER_PAIR_NUM_RETURNED=3
PRIMER_PAIR_0_PENALTY=2.035489
PRIMER_LEFT_0_PENALTY=1.938159
PRIMER_RIGHT_0_PENALTY=0.097330
PRIMER_LEFT_0_SEQUENCE=atggatgccatcaatgacctc
PRIMER_RIGHT_0_SEQUENCE=aaaggtgtcccagttgacga
PRIMER_LEFT_0=15,21
PRIMER_RIGHT_0=137,20
PRIMER_LEFT_0_TM=59.062
PRIMER_RIGHT_0_TM=60.097
PRIMER_LEFT_0_GC_PERCENT=47.619
PRIMER_RIGHT_0_GC_PERCENT=50.000
PRIMER_LEFT_0_SELF_ANY_TH=7.69
PRIMER_RIGHT_0_SELF_ANY_TH=0.00
PRIMER_LEFT_0_SELF_END_TH=0.00
PRIMER_RIGHT_0_SELF_END_TH=0.00
PRIMER_LEFT_0_HAIRPIN_TH=41.92
PRIMER_RIGHT_0_HAIRPIN_TH=38.67
PRIMER_LEFT_0_END_STABILITY=3.8500
PRIMER_RIGHT_0_END_STABILITY=4.2000
PRIMER_PAIR_0_COMPL_ANY_TH=0.00
PRIMER_PAIR_0_COMPL_END_TH=0.00
PRIMER_PAIR_0_PRODUCT_SIZE=123
PRIMER_PAIR_0_PRODUCT_TM=86.9
PRIMER_PAIR_1_PENALTY=2.221173
PRIMER_LEFT_1_PENALTY=2.122953
PRIMER_RIGHT_1_PENALTY=0.098220
PRIMER_LEFT_1_SEQUENCE=atggatgccatcaatgacct
PRIMER_RIGHT_1_SEQUENCE=gtcctcgatgatggtgtcca
PRIMER_LEFT_1=15,20
PRIMER_RIGHT_1=188,20
PRIMER_LEFT_1_TM=57.877
PRIMER_RIGHT_1_TM=60.098
PRIMER_LEFT_1_GC_PERCENT=45.000
PRIMER_RIGHT_1_GC_PERCENT=55.000
PRIMER_LEFT_1_SELF_ANY_TH=7.69
PRIMER_RIGHT_1_SELF_ANY_TH=0.00
PRIMER_LEFT_1_SELF_END_TH=0.00
PRIMER_RIGHT_1_SELF_END_TH=0.00
PRIMER_LEFT_1_HAIRPIN_TH=41.92
PRIMER_RIGHT_1_HAIRPIN_TH=38.57
PRIMER_LEFT_1_END_STABILITY=3.8500
PRIMER_RIGHT_1_END_STABILITY=4.0200
PRIMER_PAIR_1_COMPL_ANY_TH=20.44
PRIMER_PAIR_1_COMPL_END_TH=20.44
PRIMER_PAIR_1_PRODUCT_SIZE=174
PRIMER_PAIR_1_PRODUCT_TM=88.5
PRIMER_PAIR_2_PENALTY=3.536430
PRIMER_LEFT_2_PENALTY=3.438211
PRIMER_RIGHT_2_PENALTY=0.098220
PRIMER_LEFT_2_SEQUENCE=atggatgccatcaatgacctcc
PRIMER_RIGHT_2_SEQUENCE=tgtcctcgatgatggtgtcc
PRIMER_LEFT_2=15,22
PRIMER_RIGHT_2=189,20
PRIMER_LEFT_2_TM=61.438
PRIMER_RIGHT_2_TM=60.098
PRIMER_LEFT_2_GC_PERCENT=50.000
PRIMER_RIGHT_2_GC_PERCENT=55.000
PRIMER_LEFT_2_SELF_ANY_TH=7.69
PRIMER_RIGHT_2_SELF_ANY_TH=0.00
PRIMER_LEFT_2_SELF_END_TH=0.00
PRIMER_RIGHT_2_SELF_END_TH=0.00
PRIMER_LEFT_2_HAIRPIN_TH=41.92
PRIMER_RIGHT_2_HAIRPIN_TH=0.00
PRIMER_LEFT_2_END_STABILITY=4.3000
PRIMER_RIGHT_2_END_STABILITY=4.0200
PRIMER_PAIR_2_COMPL_ANY_TH=15.21
PRIMER_PAIR_2_COMPL_END_TH=15.21
PRIMER_PAIR_2_PRODUCT_SIZE=175
PRIMER_PAIR_2_PRODUCT_TM=88.4
=
