LOCUS       pSG15:_P8U17C41T13v1-1        3395 bp    DNA     circular UNA 15-AUG-2017
DEFINITION  pBAD130-tetO111213-BCD2-deGFP-T13-v1-1.
ACCESSION   urn.local.turbidostat.1j-7xtygyd
VERSION     urn.local.turbidostat.1j-7xtygyd
KEYWORDS    .
SOURCE      
  ORGANISM  
            .
FEATURES             Location/Qualifiers
     ligation        1..4
                     /label="Ligation"
     misc_feature    <1..4
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     misc_feature    5..176
                     /Source="referenceToDocument:
                     urn:local:shaobinguo:1404168489878.289"
                     /label="Digestion of IA_pBAD130-tetO-111213 gBlock by BsaI
                     - Fragment 2 (concatenated sequence 1)"
                     /note="Geneious type: Concatenated sequence"
     promoter        25..154
                     /created_by="User"
                     /label="pBAD130-tetO-101013"
     -35             53..58
                     /created_by="User"
                     /label="-35"
     operator        62..81
                     /created_by="User"
                     /label="araI1"
     -10             76..84
                     /created_by="User"
                     /label="-10"
     operator        84..101
                     /created_by="User"
                     /label="araI2"
     misc_feature    101^102
                     /Original_Bases="TCCCTATCAGTGATAGAGAT"
                     /label="TCCCTATCAGTGATAGAGAT"
                     /note="Geneious type: Editing History Deletion"
     operator        155..174
                     /created_by="User"
                     /label="tetO"
     ligation        177..180
                     /label="Ligation"
     misc_feature    181..264
                     /Source="referenceToDocument:
                     urn:local:shaobinguo:1404168028232.39"
                     /label="Digestion of IA_U17 by BsaI - Fragment 2
                     (concatenated sequence 2)"
                     /note="Geneious type: Concatenated sequence"
     misc_feature    251..>264
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     ligation        265..268
                     /label="Ligation"
     misc_feature    <265..268
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     misc_feature    269..943
                     /Source="referenceToDocument:
                     urn:local:shaobinguo:1404168028210.32"
                     /label="Digestion of IA_C41 by BsaI - Fragment 2
                     (concatenated sequence 3)"
                     /note="Geneious type: Concatenated sequence"
     PCR_product     <269..>943
                     /pair="Pair 3"
                     /set="1200bp_amp"
                     /current=0
                     /score="0.264"
                     /modified_by="User"
                     /label="1319bp-linear fragment"
     ORF             <269..>943
                     /Genetic_code="Standard"
                     /Reading_frame=3
                     /created_by="ORF Finder"
                     /label="ORF 4 (frame 3)"
     ORF             complement(<269..>943)
                     /Genetic_code="Standard"
                     /Reading_frame=3
                     /created_by="ORF Finder"
                     /label="ORF 9 (frame 3)"
     source          <269..>943
                     /dnas_title="pBEST-OR2-OR1-Pr-UTR1-deGFP-T500"
                     /label="source"
     misc_feature    <269..943
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /label="deGFP"
     ORF             <269..943
                     /Genetic_code="Standard"
                     /Reading_frame=3
                     /created_by="ORF Finder"
                     /label="ORF 2 (frame 3)"
     PCR_product     <269..>943
                     /pair="Pair 1"
                     /set="800bp_amp"
                     /current=1
                     /score="0.000"
                     /modified_by="User"
                     /label="868bp-linear fragment"
     PCR_product     <269..>943
                     /pair="Pair 1"
                     /set="1600bp_amp"
                     /current=1
                     /score="1.000"
                     /modified_by="User"
                     /label="1768bp-linear fragment"
     ORF             <269..>943
                     /Genetic_code="Standard"
                     /Reading_frame=2
                     /created_by="ORF Finder"
                     /label="ORF 1 (frame 2)"
     PCR_primer      complement(285..302)
                     /created_by="User"
                     /label="ZS2091r"
     primer_bind     599..618
                     /created_by="primer3"
                     /%GC="70.0"
                     /Tm="59.98"
                     /Hairpin="6.0"
                     /Primer-Dimer="2.0"
                     /Monovalent_Salt_Concentration="50.0"
                     /Divalent_Salt_Concentration="0.0"
                     /DNTP_Concentration="0.0"
                     /Sequence="GGCGACACCCTGGTGAACCG"
                     /annotation_group="PrimerPlugin
                     Group7d1ca663-1fe3-43a5-8c5a-3bc75c428c59_urn:local:Zachar
                     y:1339793905171.9"
                     /label="1st forward primer"
     primer_bind     682..701
                     /created_by="User"
                     /label="30424a"
     primer_bind     682..701
                     /created_by="User"
                     /label="30424b"
     ORF             complement(747..>943)
                     /Genetic_code="Standard"
                     /Reading_frame=2
                     /created_by="ORF Finder"
                     /label="ORF 8 (frame 2)"
     misc_feature    complement(757..774)
                     /created_by="User"
                     /label="30425a"
                     /note="Geneious type: primer_bind_reverse"
     primer_bind     complement(929..943)
                     /NEB_tm=64
                     /created_by="User"
                     /modified_by="User"
                     /label="ZS3022r"
     ligation        944..947
                     /label="Ligation"
     misc_feature    <944..949
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     misc_feature    948..1047
                     /Source="referenceToDocument:
                     urn:local:shaobinguo:1404168028229.37"
                     /label="Digestion of IA_T13 by BsaI - Fragment 2
                     (concatenated sequence 4)"
                     /note="Geneious type: Concatenated sequence"
     terminator      950..1039
                     /created_by="User"
                     /label="ECK120029600"
     misc_feature    1040..>1047
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     ligation        1048..1051
                     /label="Ligation"
     misc_feature    <1048..1063
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     misc_feature    1052..3395
                     /Source="referenceToDocument:
                     urn:local:shaobinguo:1404168028236.42"
                     /label="Digestion of IA_v1-1 by BsaI - Fragment 2
                     (concatenated sequence 5)"
                     /note="Geneious type: Concatenated sequence"
     PCR_product     join(3168..>3383,<1064..1284)
                     /pair="Pair 3"
                     /set="1200bp_amp"
                     /current=0
                     /score="0.264"
                     /modified_by="User"
                     /label="1319bp-linear fragment"
     source          <1064..>3383
                     /dnas_title="pBEST-OR2-OR1-Pr-UTR1-deGFP-T500"
                     /label="source"
     PCR_product     join(2924..>3383,<1064..1511)
                     /pair="Pair 1"
                     /set="1600bp_amp"
                     /current=1
                     /score="1.000"
                     /modified_by="User"
                     /label="1768bp-linear fragment"
     conflict        1064..1088
                     /created_by="User"
                     /label="area for IA_v1, v2"
     primer_bind     1076..1095
                     /created_by="primer3"
                     /%GC="70.0"
                     /Tm="59.97"
                     /Hairpin="4.0"
                     /Primer-Dimer="2.0"
                     /Monovalent_Salt_Concentration="50.0"
                     /Divalent_Salt_Concentration="0.0"
                     /DNTP_Concentration="0.0"
                     /Sequence="GGGCGCGGGGCATGACTATC"
                     /annotation_group="PrimerPlugin
                     Groupd15da267-d8a2-4186-95fc-cf70491eedb3_urn:local:Zachar
                     y:1339793905171.9"
                     /label="4th forward primer"
     misc_feature    complement(1101..1127)
                     /NEB_Tm=64
                     /created_by="User"
                     /label="ZS3036r"
                     /note="Geneious type: primer_bind_reverse"
     modified_base   1116
                     /created_by="User"
                     /label="c->g"
     misc_feature    1116
                     /Original_Bases="C"
                     /label="C"
                     /note="Geneious type: Editing History Replacement"
     primer_bind     1231..1250
                     /created_by="User"
                     /label="30424b"
     misc_feature    complement(1260..1277)
                     /NEB_Tm=60
                     /created_by="User"
                     /label="ZS3037r"
                     /note="Geneious type: primer_bind_reverse"
     PCR_primer      complement(1261..1284)
                     /dnas_title="CACATGTTCTTTCCTGCGTTATCC"
                     /primer="CACATGTTCTTTCCTGCGTTATCC"
                     /set="1200bp_amp"
                     /pair="Pair 3"
                     /current=0
                     /label="PCR primer"
     misc_feature    complement(1302..1321)
                     /created_by="User"
                     /label="30425b"
                     /note="Geneious type: primer_bind_reverse"
     rep_origin      complement(1321..2003)
                     /ApEinfo_fwdcolor="gray50"
                     /ApEinfo_revcolor="gray50"
                     /label="ColE1 origin"
     primer_bind     1361..1380
                     /created_by="primer3"
                     /annotation_group="139769645922070900032f726cc-569b-408c-9
                     fdf-83e0dc984e98"
                     /%GC="45.0"
                     /Tm="55.8"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Pair_Dimer_Tm="10.0"
                     /Product_Size=500
                     /Sequence="GACGAGCATCACAAAAATCG"
                     /label="326 F"
     primer_bind     1362..1381
                     /created_by="primer3"
                     /annotation_group="1397696459225598000e24b160f-bd90-4f3d-8
                     23a-b853eef66643"
                     /%GC="40.0"
                     /Tm="56.1"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Pair_Dimer_Tm="10.0"
                     /Product_Size=500
                     /Sequence="ACGAGCATCACAAAAATCGA"
                     /label="327 F"
     PCR_primer      complement(1488..1511)
                     /dnas_title="AAGGGAGAAAGGCGGACAGGTATC"
                     /primer="AAGGGAGAAAGGCGGACAGGTATC"
                     /set="1600bp_amp"
                     /pair="Pair 1"
                     /current=1
                     /label="PCR primer"
     misc_feature    complement(1513..1527)
                     /NEB_tm=67
                     /created_by="User"
                     /label="ZS3038r"
                     /note="Geneious type: primer_bind_reverse"
     misc_feature    1533^1534
                     /Original_Bases="A"
                     /label="A"
                     /note="Geneious type: Editing History Deletion"
     misc_feature    1535
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     primer_bind     1727..1746
                     /created_by="User"
                     /label="30424c"
     misc_feature    1780
                     /Original_Bases="G"
                     /label="G"
                     /note="Geneious type: Editing History Replacement"
     misc_feature    complement(1806..1825)
                     /created_by="User"
                     /label="30425c"
                     /note="Geneious type: primer_bind_reverse"
     misc_feature    complement(1841..1860)
                     /created_by="primer3"
                     /annotation_group="139769645922070900032f726cc-569b-408c-9
                     fdf-83e0dc984e98"
                     /%GC="45.0"
                     /Tm="56.9"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Pair_Dimer_Tm="10.0"
                     /Product_Size=500
                     /Sequence="GTTTGTTTGCCGGATCAAGA"
                     /label="825 R"
                     /note="Geneious type: primer_bind_reverse"
     misc_feature    complement(1843..1861)
                     /created_by="primer3"
                     /annotation_group="1397696459225598000e24b160f-bd90-4f3d-8
                     23a-b853eef66643"
                     /%GC="47.4"
                     /Tm="56.7"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Pair_Dimer_Tm="10.0"
                     /Product_Size=500
                     /Sequence="GGTTTGTTTGCCGGATCAA"
                     /label="826 R"
                     /note="Geneious type: primer_bind_reverse"
     CDS             complement(2098..2958)
                     /ApEinfo_fwdcolor="yellow"
                     /ApEinfo_revcolor="yellow"
                     /modified_by="User"
                     /label="AmpR"
     primer_bind     2228..2246
                     /created_by="User"
                     /label="30424d"
     modified_base   2239
                     /created_by="User"
                     /label="a->t"
     misc_feature    2239
                     /Original_Bases="A"
                     /label="A"
                     /note="Geneious type: Editing History Replacement"
     misc_feature    complement(2308..2327)
                     /created_by="User"
                     /label="30425d"
                     /note="Geneious type: primer_bind_reverse"
     misc_feature    2489
                     /Original_Bases="A"
                     /label="A"
                     /note="Geneious type: Editing History Replacement"
     misc_feature    2489
                     /created_by="User"
                     /label="A->G unique mut. not inherited from 21p"
     primer_bind     2692..2711
                     /created_by="primer3"
                     /%GC="65.0"
                     /Tm="59.97"
                     /Hairpin="5.0"
                     /Primer-Dimer="1.0"
                     /Monovalent_Salt_Concentration="50.0"
                     /Divalent_Salt_Concentration="0.0"
                     /DNTP_Concentration="0.0"
                     /Sequence="GAGTTGCTCTTGCCCGGCGT"
                     /annotation_group="PrimerPlugin
                     Group35e94697-015b-4ba9-a196-d1b75a060cac_urn:local:Zachar
                     y:1339793905171.9"
                     /label="3rd forward primer"
     primer_bind     2693..2712
                     /created_by="primer3"
                     /%GC="65.0"
                     /Tm="59.97"
                     /Hairpin="5.0"
                     /Primer-Dimer="2.0"
                     /Monovalent_Salt_Concentration="50.0"
                     /Divalent_Salt_Concentration="0.0"
                     /DNTP_Concentration="0.0"
                     /Sequence="AGTTGCTCTTGCCCGGCGTC"
                     /annotation_group="PrimerPlugin
                     Group2b969374-cce2-477b-82d4-77ad94a045e5_urn:local:Zachar
                     y:1339793905171.9"
                     /label="2nd forward primer"
     misc_feature    2715
                     /created_by="User"
                     /label="nonsilenced mutation Ile -> Val"
     misc_feature    2715
                     /Original_Bases="T"
                     /label="T"
                     /note="Geneious type: Editing History Replacement"
     primer_bind     2729..2748
                     /created_by="User"
                     /label="30424e"
     misc_feature    complement(2804..2823)
                     /created_by="User"
                     /label="30425e"
                     /note="Geneious type: primer_bind_reverse"
     primer_bind     2920..2937
                     /NEB_Tm=63
                     /created_by="User"
                     /label="ZS3038f"
     PCR_primer      2924..2946
                     /dnas_title="GGAATAAGGGCGACACGGAAATG"
                     /primer="GGAATAAGGGCGACACGGAAATG"
                     /set="1600bp_amp"
                     /pair="Pair 1"
                     /current=1
                     /label="PCR primer"
     modified_base   3158
                     /created_by="User"
                     /label="c->g"
     misc_feature    3158
                     /Original_Bases="C"
                     /label="C"
                     /note="Geneious type: Editing History Replacement"
     PCR_primer      3168..3186
                     /dnas_title="TCTGGCGAATCCTCTGACC"
                     /primer="TCTGGCGAATCCTCTGACC"
                     /set="1200bp_amp"
                     /pair="Pair 3"
                     /current=0
                     /label="PCR primer"
     primer_bind     3170..3186
                     /NEB_tm=63
                     /created_by="User"
                     /label="ZS3037f"
     primer_bind     3228..3246
                     /created_by="User"
                     /label="30424f"
     misc_feature    3238^3239
                     /Original_Bases="ACCGGATGCTGCAATTCAGAGCG"
                     /label="ACCGGATGCTGCAATTCAGAGCG"
                     /note="Geneious type: Editing History Deletion"
     modified_base   3260
                     /created_by="User"
                     /label="a->t"
     misc_feature    3260
                     /Original_Bases="A"
                     /label="A"
                     /note="Geneious type: Editing History Replacement"
     primer_bind     3290..3309
                     /NEB_Tm=61
                     /created_by="User"
                     /modified_by="User"
                     /label="ZS21111f"
     misc_feature    3298
                     /Original_Bases="A"
                     /label="A"
                     /note="Geneious type: Editing History Replacement"
     modified_base   3298
                     /created_by="User"
                     /modified_by="User"
                     /label="a->t"
     misc_feature    complement(3310..3329)
                     /neb_tm=64
                     /designed_by_primer3
                     /created_by="User"
                     /label="ZS21110r"
                     /note="Geneious type: primer_bind_reverse"
     misc_feature    complement(3310..3329)
                     /created_by="User"
                     /label="30425f"
                     /note="Geneious type: primer_bind_reverse"
     primer_bind     3320..3337
                     /NEb_Tm=60
                     /created_by="User"
                     /label="ZS3036f"
     conflict        3366..3383
                     /created_by="User"
                     /label="area for IA_vA1, IA_vA2"
     misc_feature    3384..>3395
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
ORIGIN      
        1 gcattgctgt tccgctgggc atgcacattg attatttgca cggcgtcaca ctttgctatg
       61 ccatagcatt tttatccata agattagcgg atcctacctg acgcttttta tcgcaactct
      121 ctactgtttc tccataccgt ttttttgggc tagctcccta tcagtgatag agatgcaagc
      181 gggcccaagt tcacttaaaa aggagatcaa caatgaaagc aattttcgta ctgaaacatc
      241 ttaatcatgc taaggaggtt ttctaatgga gcttttcact ggcgttgttc ccatcctggt
      301 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
      361 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
      421 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
      481 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
      541 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
      601 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
      661 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
      721 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
      781 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
      841 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
      901 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc taatgaactt tcagccaaaa
      961 aacttaagac cgccggtctt gtccactacc ttgcagtaat gcggtggaca ggatcggcgg
     1021 ttttcttttc tcttctcaac cttgagagtc gggcattgtc ttcgctcctt ccggtgggcg
     1081 cggggcatga ctatcgtcgc cgcacttatg actgtgttct ttatcatgca actcgtagga
     1141 caggtgccgg cagcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     1201 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     1261 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     1321 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     1381 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     1441 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     1501 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     1561 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     1621 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     1681 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     1741 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     1801 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     1861 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     1921 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     1981 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     2041 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     2101 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     2161 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     2221 tgctgcaatg ataccgcgtg acccacgctc accggctcca gatttatcag caataaacca
     2281 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     2341 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     2401 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     2461 ctccggttcc caacgatcaa ggcgagttgc atgatccccc atgttgtgca aaaaagcggt
     2521 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     2581 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     2641 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     2701 ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     2761 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     2821 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     2881 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     2941 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     3001 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     3061 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     3121 aacctataaa aataggcgta tcacgaggcc ctttcgtgtt caagaattct ggcgaatcct
     3181 ctgaccagcc agaaaacgac ctttctgtgg tgaaaccgga tgctgcaatt cagagcgcca
     3241 gcaagtgggg gacagcagat gacctgaccg ccgcagagtg gatgtttgac atggtgatga
     3301 ctatcgcacc atcagccaga aaaccgaatt ttgctgggtg ggctaacgat atccgcctga
     3361 tgcgtgaacg tgacggacgt aacgaagaca accac
//
