P3_SETTINGS_FILE_USED=../settings_files/primer3web_v0_4_0_default_settings.txt
P3_FILE_TYPE=settings
P3_FILE_ID=Default settings of primer3web version 0.4.0
PRIMER_THERMODYNAMIC_OLIGO_ALIGNMENT=0
PRIMER_THERMODYNAMIC_TEMPLATE_ALIGNMENT=0
PRIMER_TASK=pick_detection_primers
PRIMER_PICK_LEFT_PRIMER=1
PRIMER_PICK_INTERNAL_OLIGO=0
PRIMER_PICK_RIGHT_PRIMER=1
PRIMER_NUM_RETURN=5
PRIMER_MIN_5_PRIME_OVERLAP_OF_JUNCTION=5
PRIMER_PRODUCT_SIZE_RANGE=150-250 100-300 301-400 401-500 501-600 601-700 701-850 851-1000
PRIMER_PRODUCT_OPT_SIZE=0
PRIMER_PAIR_WT_PRODUCT_SIZE_LT=0.0
PRIMER_PAIR_WT_PRODUCT_SIZE_GT=0.0
PRIMER_MIN_SIZE=18
PRIMER_INTERNAL_MIN_SIZE=18
PRIMER_OPT_SIZE=20
PRIMER_INTERNAL_OPT_SIZE=20
PRIMER_MAX_SIZE=27
PRIMER_INTERNAL_MAX_SIZE=27
PRIMER_WT_SIZE_LT=1.0
PRIMER_INTERNAL_WT_SIZE_LT=1.0
PRIMER_WT_SIZE_GT=1.0
PRIMER_INTERNAL_WT_SIZE_GT=1.0
PRIMER_MIN_GC=20.0
PRIMER_INTERNAL_MIN_GC=20.0
PRIMER_MAX_GC=80.0
PRIMER_INTERNAL_MAX_GC=80.0
PRIMER_WT_GC_PERCENT_LT=0.0
PRIMER_INTERNAL_WT_GC_PERCENT_LT=0.0
PRIMER_WT_GC_PERCENT_GT=0.0
PRIMER_INTERNAL_WT_GC_PERCENT_GT=0.0
PRIMER_GC_CLAMP=0
PRIMER_MAX_END_GC=5
PRIMER_MIN_TM=57.0
PRIMER_INTERNAL_MIN_TM=57.0
PRIMER_OPT_TM=60.0
PRIMER_INTERNAL_OPT_TM=60.0
PRIMER_MAX_TM=63.0
PRIMER_INTERNAL_MAX_TM=63.0
PRIMER_PAIR_MAX_DIFF_TM=100.0
PRIMER_WT_TM_LT=1.0
PRIMER_INTERNAL_WT_TM_LT=1.0
PRIMER_WT_TM_GT=1.0
PRIMER_INTERNAL_WT_TM_GT=1.0
PRIMER_PAIR_WT_DIFF_TM=0.0
PRIMER_PRODUCT_MIN_TM=-1000000.0
PRIMER_PRODUCT_OPT_TM=0.0
PRIMER_PRODUCT_MAX_TM=1000000.0
PRIMER_PAIR_WT_PRODUCT_TM_LT=0.0
PRIMER_PAIR_WT_PRODUCT_TM_GT=0.0
PRIMER_TM_FORMULA=0
PRIMER_SALT_MONOVALENT=50.0
PRIMER_INTERNAL_SALT_MONOVALENT=50.0
PRIMER_SALT_DIVALENT=0.0
PRIMER_INTERNAL_SALT_DIVALENT=0.0
PRIMER_DNTP_CONC=0.0
PRIMER_INTERNAL_DNTP_CONC=0.0
PRIMER_SALT_CORRECTIONS=0
PRIMER_DNA_CONC=50.0
PRIMER_INTERNAL_DNA_CONC=50.0
PRIMER_MAX_SELF_ANY=8.00
PRIMER_INTERNAL_MAX_SELF_ANY=12.00
PRIMER_PAIR_MAX_COMPL_ANY=8.00
PRIMER_WT_SELF_ANY=0.0
PRIMER_INTERNAL_WT_SELF_ANY=0.0
PRIMER_PAIR_WT_COMPL_ANY=0.0
PRIMER_MAX_SELF_END=3.00
PRIMER_INTERNAL_MAX_SELF_END=12.00
PRIMER_PAIR_MAX_COMPL_END=3.00
PRIMER_WT_SELF_END=0.0
PRIMER_INTERNAL_WT_SELF_END=0.0
PRIMER_PAIR_WT_COMPL_END=0.0
PRIMER_MAX_END_STABILITY=9.0
PRIMER_WT_END_STABILITY=0.0
PRIMER_MAX_NS_ACCEPTED=0
PRIMER_INTERNAL_MAX_NS_ACCEPTED=0
PRIMER_WT_NUM_NS=0.0
PRIMER_INTERNAL_WT_NUM_NS=0.0
PRIMER_MAX_POLY_X=5
PRIMER_INTERNAL_MAX_POLY_X=5
PRIMER_MIN_THREE_PRIME_DISTANCE=-1
PRIMER_PICK_ANYWAY=1
PRIMER_LOWERCASE_MASKING=0
PRIMER_EXPLAIN_FLAG=1
PRIMER_LIBERAL_BASE=1
PRIMER_FIRST_BASE_INDEX=1
PRIMER_MAX_TEMPLATE_MISPRIMING=12.00
PRIMER_PAIR_MAX_TEMPLATE_MISPRIMING=24.00
PRIMER_WT_TEMPLATE_MISPRIMING=0.0
PRIMER_PAIR_WT_TEMPLATE_MISPRIMING=0.0
PRIMER_LIB_AMBIGUITY_CODES_CONSENSUS=0
PRIMER_MAX_LIBRARY_MISPRIMING=12.00
PRIMER_INTERNAL_MAX_LIBRARY_MISHYB=12.00
PRIMER_PAIR_MAX_LIBRARY_MISPRIMING=24.00
PRIMER_WT_LIBRARY_MISPRIMING=0.0
PRIMER_INTERNAL_WT_LIBRARY_MISHYB=0.0
PRIMER_PAIR_WT_LIBRARY_MISPRIMING=0.0
PRIMER_MIN_QUALITY=0
PRIMER_INTERNAL_MIN_QUALITY=0
PRIMER_MIN_END_QUALITY=0
PRIMER_QUALITY_RANGE_MIN=0
PRIMER_QUALITY_RANGE_MAX=100
PRIMER_WT_SEQ_QUAL=0.0
PRIMER_INTERNAL_WT_SEQ_QUAL=0.0
PRIMER_PAIR_WT_PR_PENALTY=1.0
PRIMER_PAIR_WT_IO_PENALTY=0.0
PRIMER_INSIDE_PENALTY=-1.0
PRIMER_OUTSIDE_PENALTY=0.0
PRIMER_WT_POS_PENALTY=0.0
PRIMER_SEQUENCING_LEAD=50
PRIMER_SEQUENCING_SPACING=500
PRIMER_SEQUENCING_INTERVAL=250
PRIMER_SEQUENCING_ACCURACY=20
PRIMER_WT_END_QUAL=0.0
PRIMER_INTERNAL_WT_END_QUAL=0.0
P3_SETTINGS_FILE_END=
SEQUENCE_ID=test_settings
SEQUENCE_TEMPLATE=atggatttgatggacatggatgccatcaatgacctccacttctctggacacagctccctcaccaacacccctacttccgacgaggattatggttgcacctggaaccactggtctcccatcgtcaactgggacacctttacaggtgccccagatgactttcaccacctcatggacaccatcatcgaggacagaacaactgtcctcgaacagctcagtccatctatcaccacaaccaccaccaccaccacaaccacggatgaggaagaagaagaaatggaaacaacaaccacaacaacaacaacggcaataaaaacgcatgaagttggtgatgattcaaaagggctaaaactagtgcacctattgatggctggcgcggaagccttaaccggttcaaccaaaaaccgtgacttagctcgagtgatattgatccggctcaaggaattagtctcacaacatgctaacggctccaacatggagagacttgccgctcatttcaccgaagcccttcatggactactagaaggtgctgggggtgcgcataataaccaccatcaccacaacaacaacaaacattacctcaccacaaatggtccccacgacaaccaaaacgacacacttgctgctttccaactacttcaagacatgtcaccttacgtcaaattcggccacttcacagccaatcaagccatcatcgaagccgtggcccacgaacgccgcgtccatgtcatcgactacgatatcatggaaggggtccaatgggcctcattaatccaatcacttgcttccaacaacaacggtccacatcttcggataaccgcattatcgcggacaggaaccggccggaggtcaatcgccaccgtacaagaaaccggcaggcgattaacctcctttgccgcttccctcggacaaccattttcttttcatcactgcagattagactccgatgaaactttccgtccttctgcattaaagcttgtacgtggagaggctttggttttcaactgcatgctgaatttacctcaccttagttaccgtgcaccggaatcagttgcttcgtttttaaacggagcaaaaacgttgaatccaaagcttgtgacgttggttgaagaagaagttggttctgttattggtgggtttgtggaaaggttcatggactcacttcatcattattcagcggtttttgattcattggaggctggttttccgatgcagaaccgggcccggactttggtggagagggtattttttgggcctagaattgcaggctcgttgggccggatatatagaacgggtggtgaagaggagagaaggtcatggggggagtggttaggtgaggtagggtttaggggagttccggtgagctttgcaaatcattgtcaagcaaagctgttgttagggctttttaatgatgggtatagggttgaagaggtgggtgtgggtagtaataagttggtgttggattggaaatcaagacgtttgctttctgcttccctttggacttgttcttcttcagattctgatttatag
PRIMER_LEFT_EXPLAIN=considered 12229, GC content failed 3, low tm 2664, high tm 6186, long poly-x seq 38, high 3' stability 222, ok 3116
PRIMER_RIGHT_EXPLAIN=considered 12305, GC content failed 3, low tm 2880, high tm 5939, long poly-x seq 43, high 3' stability 212, ok 3228
PRIMER_PAIR_EXPLAIN=considered 60, unacceptable product size 54, ok 6
PRIMER_LEFT_NUM_RETURNED=5
PRIMER_RIGHT_NUM_RETURNED=5
PRIMER_INTERNAL_NUM_RETURNED=0
PRIMER_PAIR_NUM_RETURNED=5
PRIMER_PAIR_0_PENALTY=0.037132
PRIMER_LEFT_0_PENALTY=0.031043
PRIMER_RIGHT_0_PENALTY=0.006089
PRIMER_LEFT_0_SEQUENCE=actggtctcccatcgtcaac
PRIMER_RIGHT_0_SEQUENCE=gcgtttttattgccgttgtt
PRIMER_LEFT_0=107,20
PRIMER_RIGHT_0=316,20
PRIMER_LEFT_0_TM=59.969
PRIMER_RIGHT_0_TM=60.006
PRIMER_LEFT_0_GC_PERCENT=55.000
PRIMER_RIGHT_0_GC_PERCENT=40.000
PRIMER_LEFT_0_SELF_ANY=3.00
PRIMER_RIGHT_0_SELF_ANY=2.00
PRIMER_LEFT_0_SELF_END=2.00
PRIMER_RIGHT_0_SELF_END=0.00
PRIMER_LEFT_0_END_STABILITY=6.7000
PRIMER_RIGHT_0_END_STABILITY=7.0000
PRIMER_LEFT_0_TEMPLATE_MISPRIMING=10.0000
PRIMER_RIGHT_0_TEMPLATE_MISPRIMING=8.0000
PRIMER_PAIR_0_COMPL_ANY=5.00
PRIMER_PAIR_0_COMPL_END=3.00
PRIMER_PAIR_0_PRODUCT_SIZE=210
PRIMER_PAIR_0_PRODUCT_TM=77.4
PRIMER_PAIR_0_TEMPLATE_MISPRIMING=17.00
PRIMER_PAIR_1_PENALTY=0.050921
PRIMER_LEFT_1_PENALTY=0.044832
PRIMER_RIGHT_1_PENALTY=0.006089
PRIMER_LEFT_1_SEQUENCE=cgaggattatggttgcacct
PRIMER_RIGHT_1_SEQUENCE=gcgtttttattgccgttgtt
PRIMER_LEFT_1=81,20
PRIMER_RIGHT_1=316,20
PRIMER_LEFT_1_TM=59.955
PRIMER_RIGHT_1_TM=60.006
PRIMER_LEFT_1_GC_PERCENT=50.000
PRIMER_RIGHT_1_GC_PERCENT=40.000
PRIMER_LEFT_1_SELF_ANY=4.00
PRIMER_RIGHT_1_SELF_ANY=2.00
PRIMER_LEFT_1_SELF_END=2.00
PRIMER_RIGHT_1_SELF_END=0.00
PRIMER_LEFT_1_END_STABILITY=7.9000
PRIMER_RIGHT_1_END_STABILITY=7.0000
PRIMER_LEFT_1_TEMPLATE_MISPRIMING=9.0000
PRIMER_RIGHT_1_TEMPLATE_MISPRIMING=8.0000
PRIMER_PAIR_1_COMPL_ANY=3.00
PRIMER_PAIR_1_COMPL_END=0.00
PRIMER_PAIR_1_PRODUCT_SIZE=236
PRIMER_PAIR_1_PRODUCT_TM=77.9
PRIMER_PAIR_1_TEMPLATE_MISPRIMING=17.00
PRIMER_PAIR_2_PENALTY=0.052224
PRIMER_LEFT_2_PENALTY=0.000037
PRIMER_RIGHT_2_PENALTY=0.052187
PRIMER_LEFT_2_SEQUENCE=atcgccaccgtacaagaaac
PRIMER_RIGHT_2_SEQUENCE=cggtgcacggtaactaaggt
PRIMER_LEFT_2=850,20
PRIMER_RIGHT_2=1041,20
PRIMER_LEFT_2_TM=60.000
PRIMER_RIGHT_2_TM=60.052
PRIMER_LEFT_2_GC_PERCENT=50.000
PRIMER_RIGHT_2_GC_PERCENT=55.000
PRIMER_LEFT_2_SELF_ANY=4.00
PRIMER_RIGHT_2_SELF_ANY=6.00
PRIMER_LEFT_2_SELF_END=0.00
PRIMER_RIGHT_2_SELF_END=3.00
PRIMER_LEFT_2_END_STABILITY=6.7000
PRIMER_RIGHT_2_END_STABILITY=7.9000
PRIMER_LEFT_2_TEMPLATE_MISPRIMING=8.0000
PRIMER_RIGHT_2_TEMPLATE_MISPRIMING=7.0000
PRIMER_PAIR_2_COMPL_ANY=6.00
PRIMER_PAIR_2_COMPL_END=2.00
PRIMER_PAIR_2_PRODUCT_SIZE=192
PRIMER_PAIR_2_PRODUCT_TM=76.4
PRIMER_PAIR_2_TEMPLATE_MISPRIMING=15.00
PRIMER_PAIR_3_PENALTY=0.065561
PRIMER_LEFT_3_PENALTY=0.006937
PRIMER_RIGHT_3_PENALTY=0.058624
PRIMER_LEFT_3_SEQUENCE=gtcctcgaacagctcagtcc
PRIMER_RIGHT_3_SEQUENCE=agccggatcaatatcactcg
PRIMER_LEFT_3=199,20
PRIMER_RIGHT_3=434,20
PRIMER_LEFT_3_TM=59.993
PRIMER_RIGHT_3_TM=60.059
PRIMER_LEFT_3_GC_PERCENT=60.000
PRIMER_RIGHT_3_GC_PERCENT=50.000
PRIMER_LEFT_3_SELF_ANY=4.00
PRIMER_RIGHT_3_SELF_ANY=4.00
PRIMER_LEFT_3_SELF_END=2.00
PRIMER_RIGHT_3_SELF_END=2.00
PRIMER_LEFT_3_END_STABILITY=7.6000
PRIMER_RIGHT_3_END_STABILITY=8.1000
PRIMER_LEFT_3_TEMPLATE_MISPRIMING=7.0000
PRIMER_RIGHT_3_TEMPLATE_MISPRIMING=8.0000
PRIMER_PAIR_3_COMPL_ANY=3.00
PRIMER_PAIR_3_COMPL_END=1.00
PRIMER_PAIR_3_PRODUCT_SIZE=236
PRIMER_PAIR_3_PRODUCT_TM=76.6
PRIMER_PAIR_3_TEMPLATE_MISPRIMING=15.00
PRIMER_PAIR_4_PENALTY=0.073199
PRIMER_LEFT_4_PENALTY=0.066262
PRIMER_RIGHT_4_PENALTY=0.006937
PRIMER_LEFT_4_SEQUENCE=gccatcaatgacctccactt
PRIMER_RIGHT_4_SEQUENCE=ggactgagctgttcgaggac
PRIMER_LEFT_4=22,20
PRIMER_RIGHT_4=218,20
PRIMER_LEFT_4_TM=59.934
PRIMER_RIGHT_4_TM=59.993
PRIMER_LEFT_4_GC_PERCENT=50.000
PRIMER_RIGHT_4_GC_PERCENT=60.000
PRIMER_LEFT_4_SELF_ANY=5.00
PRIMER_RIGHT_4_SELF_ANY=4.00
PRIMER_LEFT_4_SELF_END=0.00
PRIMER_RIGHT_4_SELF_END=2.00
PRIMER_LEFT_4_END_STABILITY=6.7000
PRIMER_RIGHT_4_END_STABILITY=7.6000
PRIMER_LEFT_4_TEMPLATE_MISPRIMING=9.0000
PRIMER_RIGHT_4_TEMPLATE_MISPRIMING=8.0000
PRIMER_PAIR_4_COMPL_ANY=4.00
PRIMER_PAIR_4_COMPL_END=1.00
PRIMER_PAIR_4_PRODUCT_SIZE=197
PRIMER_PAIR_4_PRODUCT_TM=79.1
PRIMER_PAIR_4_TEMPLATE_MISPRIMING=16.00
=
