LOCUS       pB7WGR2                11611 bp    DNA     circular     12-JUL-2022
DEFINITION  .
ACCESSION   urn.local...17r-dsop7al
KEYWORDS    .
SOURCE      
  ORGANISM  .
COMMENT     http://www.informaxinc.com/                                        
            This file is created by Vector NTI                                 
            http://www.invitrogen.com/                                         
            ORIGDB|GenBank                                                     
            VNTDATE|342126914|                                                 
            VNTDBDATE|457854192|                                               
            LSOWNER|                                                           
            VNTNAME|pB7WGR2,0|                                                 
            VNTAUTHORNAME|Demo User|                                           
            VNTAUTHOREML|manka@psb.ugent.be|                                   
            VNTOAUTHORNAME|Mansour Karimi|                                     
            VNTOAUTHORTEL|+3193313850|                                         
            VNTOAUTHOREML|mansour.karimi@psb.ugent.be|                         
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FEATURES             Location/Qualifiers
     Promoter        8..282
                     /Mismatches=0
                     /%_Identity=100
                     /Motif="CACGAACCCAGTGGACATAAGCCTGTTCGGTTCGTAAGCTGTAATGCAAG
                     TAGCGTATGCGCTCACGCAACTGGTCCAGAACCTTGACCGAACGCAGCGGTGGTAACG
                     GCGCAGTGGCGGTTTTCATGGCTTGTTATGACTGTTTTTTTGGGGTACAGTCTATGCC
                     TCGGGCATCCAAGCAGCAAGCGCGTTACGCCGTGGGTCGATGTTTGATGTTATGGAGC
                     AGCAACGATGTTACGCAGCAGGGCAGTCGCCCTAAAACAAAGTTAAACATC"
                     /X-Transferred_From="pGGZ004"
                     /X-Transferred_Similarity="100.00%"
                     /X-Primary_Match="100.00%; Adjusted-Interval-0:0/0; P1 (<a
                     href=""urn:local:maizel:2p5-ccnltde"">pGGZ004</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0; P1
                     (<a href=""urn:local:maizel:2p3-ccnltde"">pR450</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0; P1
                     (<a href=""urn:local:thjac:47-ckxljc3"">SpR</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0; P1
                     (<a
                     href=""urn:local:maizel:2oq-ccnltdd"">pR451_pFASTR_A-G</a>
                     )"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0; P1
                     (<a href=""urn:local:maizel:2p4-ccnltde"">pR449</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     pAgro1 (<a
                     href=""urn:local:mamee:3jv-esp0psa"">pGG-A-PAgro1-B</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     pAgro1 (<a
                     href=""urn:local:mamee:3k2-esp0psa"">pEN-L4-PAGRO1-SpCas9-
                     T3T-R1 </a>)"
                     /X-Alternative_Match="92.86%; Adjusted-Interval-0:12/-235;
                     Pc promoter (<a
                     href=""urn:local:mamee:405-esp0pt5"">pDEST32-GG</a>)"
                     /X-Alternative_Match="90.14%; Adjusted-Interval-0:0/0; P1
                     (<a href=""urn:local:mamee:3uc-esp0psx"">pGADT7-GG</a>)"
                     /note="Derived using Geneious Prime 2022.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="SpR"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; P1 (<a
                     href=""urn:local:.:1p-f0tk4mj"">SpR</a>)"
                     /label="P1"
     misc_feature    283..1074
                     /note="Derived using Geneious Prime 2021.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /X-Transferred_From="SpR"
                     /X-Transferred_Similarity="98.72%"
                     /X-Primary_Match="98.72%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:8l-dgycryy"">SpR</a>"
                     /modified_by="mamee"
                     /note="Derived using Geneious Prime 2022.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="SpR"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:1p-f0tk4mj"">SpR</a>"
                     /label="SpR"
                     /note="Geneious type: Bacterial Selection"
     primer_bind     complement(728..747)
                     /%GC="55.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="60.1"
                     /annotation_group="59831076898668004df777a4-e416-4333-a755
                     -33aed33d9c3e"
                     /created_by="primer3"
                     /Sequence="ATCAAAGAGTTCCTCCGCCG"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pB7WGR2"
                     /label="P7"
     rep_origin      1320..1908
                     /created_by="thjac"
                     /modified_by="mamee"
                     /note="Derived using Geneious Prime 2021.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="high-copy-number ColE1/pMB1/pBR322/pUC
                     origin of  replication"
                     /Transferred_Similarity="99.83%"
                     /Primary_Match="99.83%; Adjusted-Interval-0:0/0;
                     ColE1/pMB1/pBR322/pUC Ori (<a
                     href=""urn:local:.:8d-dk4mzop"">high-copy-number
                     ColE1/pMB1/pBR322/pUC origin of  replication</a>)"
                     /label="ColE1/pMB1/pBR322/pUC Ori"
     primer_bind     complement(1500..1519)
                     /%GC="55.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="58.9"
                     /annotation_group="5983107633032700ba0edf5c-2e6f-442e-b975
                     -917b6dd4f1cf"
                     /created_by="primer3"
                     /Sequence="GCGGTGCTACAGAGTTCTTG"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pB7WGR2"
                     /label="P6"
     regulatory      complement(1944..2204)
                     /%_Identity=100
                     /Motif="CGGAGTGTATACTGGCTTAACTATGCGGCATCAGAGCAGATTGTACTGAG
                     AGTGCACCATATGCGGTGTGAAATACCGCACAGATGCGTAAGGAGAAAATACCGCATC
                     AGGCGCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGC
                     GAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAA
                     CGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAG"
                     /annotation_group="pBR322: 7,710 -> 7,970"
                     /modified_by="User"
                     /modified_by="mamee"
                     /note="Derived using Geneious Prime 2021.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="pBR322 bom"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:7a-dlucaql"">pBR322 bom</a>"
                     /label="pBR322 bom"
     primer_bind     complement(2324..2343)
                     /%GC="55.0"
                     /Hairpin_Tm="33.5"
                     /Self_Dimer_Tm="None"
                     /Tm="60.0"
                     /annotation_group="59831075756834001d42f0ba-2b2a-4fb3-a091
                     -dece45366438"
                     /created_by="primer3"
                     /Sequence="ACCTCTGACACATGCAGCTC"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pB7WGR2"
                     /label="P5"
     rep_origin      2578..2772
                     /note="origin of replication fromÂ Pseudomonas pVS1
                     plasmid"
                     /modified_by="mamee"
                     /note="Derived using Geneious Prime 2021.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="pVS1 oriV"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:7t-dlucpl4"">pVS1 oriV</a>"
                     /label="pVS1 oriV"
     regulatory      complement(2838..3911)
                     /note="Pseudomonas pVS1 plasmid replication protein"
                     /product="replication protein RepA"
                     /gene="repA"
                     /regulatory_class="other"
                     /note="Derived using Geneious Prime 2021.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="pVS1 repA"
                     /Transferred_Similarity="99.91%"
                     /Primary_Match="99.91%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:7b-dlucaql"">pVS1 repA</a>"
                     /label="pVS1 repA"
     primer_bind     complement(3126..3143)
                     /%GC="61.1"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="0.7"
                     /Tm="60.0"
                     /annotation_group="5983107517401300d28b4127-6a27-4ebc-826a
                     -9741c07569f7"
                     /created_by="primer3"
                     /Sequence="AAGAACCCGGACGTGCTG"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pB7WGR2"
                     /label="P4"
     primer_bind     complement(3929..3947)
                     /%GC="63.2"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="61.0"
                     /annotation_group="5983107457128500be8724dc-01cd-454d-995f
                     -142adc0b18f8"
                     /created_by="primer3"
                     /Sequence="CAATGGCACTGGAACCCCC"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pB7WGR2"
                     /label="P3"
     regulatory      complement(4340..4969)
                     /note="Pseudomonas pVS1 plasmid stability protein"
                     /product="plasmid stability protein StaA"
                     /gene="staA"
                     /regulatory_class="other"
                     /note="Derived using Geneious Prime 2021.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="pVS1 staA"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:7d-dlucaql"">pVS1 staA</a>"
                     /label="pVS1 staA"
     primer_bind     complement(4725..4744)
                     /%GC="55.0"
                     /Hairpin_Tm="42.2"
                     /Self_Dimer_Tm="None"
                     /Tm="60.0"
                     /annotation_group="5983107365783100b1bcf8c0-b19c-47ef-8996
                     -ddfa6fee45f4"
                     /created_by="primer3"
                     /Sequence="CGCGACTTCGTAGTGATCGA"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pB7WGR2"
                     /label="P2"
     primer_bind     complement(5518..5536)
                     /%GC="63.2"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="61.3"
                     /annotation_group="5983107308570800548c6e1f-1ef1-4f42-89e8
                     -0b1b9312a5e4"
                     /created_by="primer3"
                     /Sequence="CCCGAGGCGTGAAGTTTGG"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pB7WGR2"
                     /label="P1"
     T-DNA_border    6155..6385
                     /modified_by="wacae"
                     /note="Derived using Geneious Prime 2022.0.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="RB"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:6f-dgycryw"">RB</a>"
                     /label="RB"
     primer_bind     complement(6345..6364)
                     /%GC="50.0"
                     /Hairpin_Tm="36.2"
                     /Self_Dimer_Tm="None"
                     /Tm="60.0"
                     /annotation_group="59831081132708003e14e87f-9178-44e2-909f
                     -a90b32b35c8c"
                     /created_by="primer3"
                     /Sequence="AGATTGTCGTTTCCCGCCTT"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pB7WGR2"
                     /label="P14"
     promoter        6387..7433
                     /modified_by="mamee"
                     /note="Derived using Geneious Prime 2022.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="pCaMV 35S - Gateway"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; p35S (<a
                     href=""urn:local:.:3a-dlt03ty"">pCaMV 35S - Gateway</a>)"
                     /label="p35S"
     primer_bind     complement(7117..7138)
                     /%GC="45.5"
                     /Hairpin_Tm="41.9"
                     /Self_Dimer_Tm="12.2"
                     /Tm="59.9"
                     /annotation_group="5983108342373000e613eb0f-dd6a-4f13-b876
                     -ab99332ad457"
                     /created_by="primer3"
                     /Sequence="CGCAATGATGGCATTTGTAGGT"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pB7WGR2"
                     /label="P18"
     CDS             7455..8129
                     /label="rfp"
     primer_bind     complement(7861..7880)
                     /%GC="55.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="11.7"
                     /Tm="60.3"
                     /annotation_group="59831084101107004cec30ce-129a-42d4-8739
                     -913b0407ff0c"
                     /created_by="primer3"
                     /Sequence="GCCCATGGTCTTCTTCTGCA"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pB7WGR2"
                     /label="P34"
     attR1           8136..8260
                     /modified_by="mamee"
                     /note="Derived using Geneious Prime 2022.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="attR1"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:1c-f0o2y6y"">attR1</a>"
     promoter        8285..8315
                     /note="Lac promoter of Escherichia coli containing 2 base
                     pair mutations"
                     /regulatory_class="promoter"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /X-Transferred_From="lacUV5"
                     /X-Transferred_Similarity="100.00%"
                     /X-Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:1g0-c949y1m"">lacUV5</a>"
                     /note="Derived using Geneious Prime 2022.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="CmR"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; lacUV5
                     (<a href=""urn:local:.:29-f0o4v7s"">CmR</a>)"
                     /label="lacUV5"
     misc_feature    8369..9028
                     /Transferred_From="cmR"
                     /Transferred_Similarity="99.85%"
                     /modified_by="mamee"
                     /note="Derived using Geneious Prime 2022.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="CmR"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:29-f0o4v7s"">CmR</a>"
                     /label="CmR"
                     /note="Geneious type: Bacterial Selection"
     primer_bind     complement(8599..8618)
                     /%GC="55.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="60.0"
                     /annotation_group="59831082280289000effee68-d2a1-44ff-a6dc
                     -56b15bcb1036"
                     /created_by="primer3"
                     /Sequence="TCACCAGCTCACCGTCTTTC"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pB7WGR2"
                     /label="P16"
     UTR             9051..9369
                     /Mismatches=1
                     /%_Identity="99.68652037617555"
                     /Motif="AAGCCAGATAACAGTATGCGTATTTGCGCGCTGATTTTTGCGGTATAAGA
                     ATATATACTGATATGTATACCCGAAGTATGTCAAAAGGAGGTATGCTATGAAGCAGCG
                     TATTACAGTGACAGTTGACAGCGACAGCTATCAGTTGCTCAAGGCATATATGATGTCA
                     ATATCTCCGGTCTGGTAAGCACAACCATGCAGAATGAAGCCCGTCGTCTGCGTGCCGA
                     ACGCTGGAAAGCGGAAAATCAGGAAGGGATGGCTGAGGTCGCCCGGTTTATTGAAATG
                     AACGGCTCTTTTGCTGACGAGAACAGGGGCTGGTGAA"
                     /note="Derived using Geneious Prime 2021.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="ccdB"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; ccdB
                     5'UTR (<a href=""urn:local:.:8r-dgycryy"">ccdB</a>)"
                     /label="ccdB 5'UTR"
     CDS             9370..9675
                     /note="Derived using Geneious Prime 2021.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="ccdB"
                     /Transferred_Similarity="99.67%"
                     /Primary_Match="99.67%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:8r-dgycryy"">ccdB</a>"
                     /label="ccdB"
     primer_bind     complement(9395..9415)
                     /%GC="47.6"
                     /Hairpin_Tm="45.0"
                     /Self_Dimer_Tm="None"
                     /Tm="57.6"
                     /annotation_group="59831081712833001f1cbfcd-e7e9-4748-8a7a
                     -14b845c9a211"
                     /created_by="primer3"
                     /Sequence="GACGATAACGGCTCTCTCTTT"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pB7WGR2"
                     /label="P15"
     attR2           complement(9716..9840)
                     /modified_by="mamee"
                     /note="Derived using Geneious Prime 2022.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="attR2"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:1j-f0o30z1"">attR2</a>"
     terminator      9850..10075
                     /created_by="mamee"
                     /note="Derived using Geneious Prime 2022.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="t35S"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:lk-f0ukkf4"">t35S</a>"
                     /label="t35S"
     primer_bind     complement(10062..10081)
                     /%GC="55.0"
                     /Hairpin_Tm="44.4"
                     /Self_Dimer_Tm="32.8"
                     /Tm="60.6"
                     /annotation_group="59831082856462000a85439d-5c35-4b26-a1e4
                     -862ade216871"
                     /created_by="primer3"
                     /Sequence="GCCTGCAGGTCACTGGATTT"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pB7WGR2"
                     /label="P17"
     promoter        10140..10412
                     /created_by="thjac"
                     /modified_by="mamee"
                     /note="Derived using Geneious Prime 2021.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /X-Transferred_From="pNOS"
                     /X-Transferred_Similarity="100.00%"
                     /X-Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:77-dgycryx"">pNOS</a>"
                     /note="Derived using Geneious Prime 2022.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="Basta"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; pNOS (<a
                     href=""urn:local:.:1k-e500sax"">Basta</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:ip-f0oyhrv"">pNOS</a>"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; pNOS
                     (<a href=""urn:local:.:lm-f0p7ifx"">Hygromycin</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; pNOS
                     (<a href=""urn:local:.:2j-e501a1r"">Kanamycin</a>)"
                     /label="pNOS"
     Plant_Selection 10140..11258
                     /modified_by="mamee"
                     /note="Derived using Geneious Prime 2022.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="Basta"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:1k-e500sax"">Basta</a>"
                     /label="Basta"
     CDS             10433..10984
                     /%_Identity=100
                     /Motif="ATGAGCCCAGAACGACGCCCGGCCGACATCCGCCGTGCCACCGAGGCGGA
                     CATGCCGGCGGTCTGCACCATCGTCAACCACTACATCGAGACAAGCACGGTCAACTTC
                     CGTACCGAGCCGCAGGAACCGCAGGAGTGGACGGACGACCTCGTCCGTCTGCGGGAGC
                     GCTATCCCTGGCTCGTCGCCGAGGTGGACGGCGAGGTCGCCGGCATCGCCTACGCGGG
                     TCCCTGGAAGGCACGCAACGCCTACGACTGGACGGCCGAGTCGACCGTGTACGTCTCC
                     CCCCGCCACCAGCGGACGGGACTGGGCTCCACGCTCTACACCCACCTGCTGAAGTCCC
                     TGGAGGCACAGGGCTTCAAGAGCGTGGTCGCTGTCATCGGGCTGCCCAACGACCCGAG
                     CGTGCGCATGCACGAGGCGCTCGGATATGCCCCCCGCGGCATGCTGCGGGCGGCCGGC
                     TTCAAGCACGGGAACTGGCATGACGTGGGTTTCTGGCAGCTGGACTTCAGCCTGCCGG
                     TGCCGCCCCGTCCGGTCCTGCCCGTCACCGAAATCTGA"
                     /annotation_group="PAT: 8,732 -> 9,283"
                     /modified_by="mamee"
                     /note="Derived using Geneious Prime 2022.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PAT"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:98-dgycryy"">PAT</a>"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; PAT
                     (<a href=""urn:local:.:1k-e500sax"">Basta</a>)"
                     /label="PAT"
     primer_bind     complement(10748..10767)
                     /%GC="55.0"
                     /Hairpin_Tm="45.2"
                     /Self_Dimer_Tm="None"
                     /Tm="59.0"
                     /annotation_group="5983107821995000b1ecd971-5052-418a-8687
                     -25ee8462bcb5"
                     /created_by="primer3"
                     /Sequence="TTCAGCAGGTGGGTGTAGAG"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pB7WGR2"
                     /label="P9"
     terminator      10991..11258
                     /note="Derived using Geneious Prime 2020.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /X-Transferred_From="NOST"
                     /X-Transferred_Similarity="100.00%"
                     /X-Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:6c-dgycryw"">NOST</a>"
                     /modified_by="mamee"
                     /note="Derived using Geneious Prime 2022.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="tNOS"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:38-f1t9off"">tNOS</a>"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; tNOS
                     (<a href=""urn:local:.:1k-e500sax"">Basta</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; tNOS
                     (<a href=""urn:local:.:lm-f0p7ifx"">Hygromycin</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; tNOS
                     (<a href=""urn:local:.:2j-e501a1r"">Kanamycin</a>)"
                     /label="tNOS"
     T-DNA_border    11279..11611
                     /modified_by="wacae"
                     /note="Derived using Geneious Prime 2021.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="LB"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:6e-dgycryw"">LB</a>"
                     /label="LB"
     primer_bind     complement(11523..11542)
                     /%GC="50.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="59.9"
                     /annotation_group="598310775462350044e70cfe-606d-4463-9897
                     -0bbccbc6a81c"
                     /created_by="primer3"
                     /Sequence="TCAAACCCGGCAGCTTAGTT"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pB7WGR2"
                     /label="P8"
ORIGIN      
        1 attcgggcac gaacccagtg gacataagcc tgttcggttc gtaagctgta atgcaagtag
       61 cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg taacggcgca
      121 gtggcggttt tcatggcttg ttatgactgt ttttttgggg tacagtctat gcctcgggca
      181 tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga tgttatggag cagcaacgat
      241 gttacgcagc agggcagtcg ccctaaaaca aagttaaaca tcatggggga agcggtgatc
      301 gccgaagtat cgactcaact atcagaggta gttggcgtca tcgagcgcca tctcgaaccg
      361 acgttgctgg ccgtacattt gtacggctcc gcagtggatg gcggcctgaa gccacacagt
      421 gatattgatt tgctggttac ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg
      481 atcaacgacc ttttggaaac ttcggcttcc cctggagaga gcgagattct ccgcgctgta
      541 gaagtcacca ttgttgtgca cgacgacatc attccgtggc gttatccagc taagcgcgaa
      601 ctgcaatttg gagaatggca gcgcaatgac attcttgcag gtatcttcga gccagccacg
      661 atcgacattg atctggctat cttgctgaca aaagcaagag aacatagcgt tgccttggta
      721 ggtccagcgg cggaggaact ctttgatccg gttcctgaac aggatctatt tgaggcgcta
      781 aatgaaacct taacgctatg gaactcgccg cccgactggg ctggcgatga gcgaaatgta
      841 gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat
      901 gtcgctgccg actgggcaat ggagcgcctg ccggcccagt atcagcccgt catacttgaa
      961 gctagacagg cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg
     1021 gaagaatttg tccactacgt gaaaggcgag atcaccaagg tagtcggcaa ataatgtcta
     1081 gctagaaatt cgttcaagcc gacgccgctt cgcggcgcgg cttaactcaa gcgttagatg
     1141 cactaagcac ataattgctc acagccaaac tatcaggtca agtctgcttt tattattttt
     1201 aagcgtgcat aataagccct acacaaattg ggagatatat catgcatgac caaaatccct
     1261 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     1321 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     1381 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     1441 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     1501 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     1561 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     1621 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     1681 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     1741 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     1801 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     1861 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     1921 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     1981 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     2041 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     2101 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt
     2161 acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact
     2221 gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     2281 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     2341 ggttttcacc gtcatcaccg aaacgcgcga ggcagggtgc cttgatgtgg gcgccggcgg
     2401 tcgagtggcg acggcgcggc ttgtccgcgc cctggtagat tgcctggccg taggccagcc
     2461 atttttgagc ggccagcggc cgcgataggc cgacgcgaag cggcggggcg tagggagcgc
     2521 agcgaccgaa gggtaggcgc tttttgcagc tcttcggctg tgcgctggcc agacagttat
     2581 gcacaggcca ggcgggtttt aagagtttta ataagtttta aagagtttta ggcggaaaaa
     2641 tcgccttttt tctcttttat atcagtcact tacatgtgtg accggttccc aatgtacggc
     2701 tttgggttcc caatgtacgg gttccggttc ccaatgtacg gctttgggtt cccaatgtac
     2761 gtgctatcca caggaaagag accttttcga cctttttccc ctgctagggc aatttgccct
     2821 agcatctgct ccgtacatta ggaaccggcg gatgcttcgc cctcgatcag gttgcggtag
     2881 cgcatgacta ggatcgggcc agcctgcccc gcctcctcct tcaaatcgta ctccggcagg
     2941 tcatttgacc cgatcagctt gcgcacggtg aaacagaact tcttgaactc tccggcgctg
     3001 ccactgcgtt cgtagatcgt cttgaacaac catctggctt ctgccttgcc tgcggcgcgg
     3061 cgtgccaggc ggtagagaaa acggccgatg ccgggatcga tcaaaaagta atcggggtga
     3121 accgtcagca cgtccgggtt cttgccttct gtgatctcgc ggtacatcca atcagctagc
     3181 tcgatctcga tgtactccgg ccgcccggtt tcgctcttta cgatcttgta gcggctaatc
     3241 aaggcttcac cctcggatac cgtcaccagg cggccgttct tggccttctt cgtacgctgc
     3301 atggcaacgt gcgtggtgtt taaccgaatg caggtttcta ccaggtcgtc tttctgcttt
     3361 ccgccatcgg ctcgccggca gaacttgagt acgtccgcaa cgtgtggacg gaacacgcgg
     3421 ccgggcttgt ctcccttccc ttcccggtat cggttcatgg attcggttag atgggaaacc
     3481 gccatcagta ccaggtcgta atcccacaca ctggccatgc cggccggccc tgcggaaacc
     3541 tctacgtgcc cgtctggaag ctcgtagcgg atcacctcgc cagctcgtcg gtcacgcttc
     3601 gacagacgga aaacggccac gtccatgatg ctgcgactat cgcgggtgcc cacgtcatag
     3661 agcatcggaa cgaaaaaatc tggttgctcg tcgcccttgg gcggcttcct aatcgacggc
     3721 gcaccggctg ccggcggttg ccgggattct ttgcggattc gatcagcggc cgcttgccac
     3781 gattcaccgg ggcgtgcttc tgcctcgatg cgttgccgct gggcggcctg cgcggccttc
     3841 aacttctcca ccaggtcatc acccagcgcc gcgccgattt gtaccgggcc ggatggtttg
     3901 cgaccgctca cgccgattcc tcgggcttgg gggttccagt gccattgcag ggccggcaga
     3961 caacccagcc gcttacgcct ggccaaccgc ccgttcctcc acacatgggg cattccacgg
     4021 cgtcggtgcc tggttgttct tgattttcca tgccgcctcc tttagccgct aaaattcatc
     4081 tactcattta ttcatttgct catttactct ggtagctgcg cgatgtattc agatagcagc
     4141 tcggtaatgg tcttgccttg gcgtaccgcg tacatcttca gcttggtgtg atcctccgcc
     4201 ggcaactgaa agttgacccg cttcatggct ggcgtgtctg ccaggctggc caacgttgca
     4261 gccttgctgc tgcgtgcgct cggacggccg gcacttagcg tgtttgtgct tttgctcatt
     4321 ttctctttac ctcattaact caaatgagtt ttgatttaat ttcagcggcc agcgcctgga
     4381 cctcgcgggc agcgtcgccc tcgggttctg attcaagaac ggttgtgccg gcggcggcag
     4441 tgcctgggta gctcacgcgc tgcgtgatac gggactcaag aatgggcagc tcgtacccgg
     4501 ccagcgcctc ggcaacctca ccgccgatgc gcgtgccttt gatcgcccgc gacacgacaa
     4561 aggccgcttg tagccttcca tccgtgacct caatgcgctg cttaaccagc tccaccaggt
     4621 cggcggtggc ccatatgtcg taagggcttg gctgcaccgg aatcagcacg aagtcggctg
     4681 ccttgatcgc ggacacagcc aagtccgccg cctggggcgc tccgtcgatc actacgaagt
     4741 cgcgccggcc gatggccttc acgtcgcggt caatcgtcgg gcggtcgatg ccgacaacgg
     4801 ttagcggttg atcttcccgc acggccgccc aatcgcgggc actgccctgg ggatcggaat
     4861 cgactaacag aacatcggcc ccggcgagtt gcagggcgcg ggctagatgg gttgcgatgg
     4921 tcgtcttgcc tgacccgcct ttctggttaa gtacagcgat aaccttcatg cgttcccctt
     4981 gcgtatttgt ttatttactc atcgcatcat atacgcagcg accgcatgac gcaagctgtt
     5041 ttactcaaat acacatcacc tttttagacg gcggcgctcg gtttcttcag cggccaagct
     5101 ggccggccag gccgccagct tggcatcaga caaaccggcc aggatttcat gcagccgcac
     5161 ggttgagacg tgcgcgggcg gctcgaacac gtacccggcc gcgatcatct ccgcctcgat
     5221 ctcttcggta atgaaaaacg gttcgtcctg gccgtcctgg tgcggtttca tgcttgttcc
     5281 tcttggcgtt cattctcggc ggccgccagg gcgtcggcct cggtcaatgc gtcctcacgg
     5341 aaggcaccgc gccgcctggc ctcggtgggc gtcacttcct cgctgcgctc aagtgcgcgg
     5401 tacagggtcg agcgatgcac gccaagcagt gcagccgcct ctttcacggt gcggccttcc
     5461 tggtcgatca gctcgcgggc gtgcgcgatc tgtgccgggg tgagggtagg gcgggggcca
     5521 aacttcacgc ctcgggcctt ggcggcctcg cgcccgctcc gggtgcggtc gatgattagg
     5581 gaacgctcga actcggcaat gccggcgaac acggtcaaca ccatgcggcc ggccggcgtg
     5641 gtggtgtcgg cccacggctc tgccaggcta cgcaggcccg cgccggcctc ctggatgcgc
     5701 tcggcaatgt ccagtaggtc gcgggtgctg cgggccaggc ggtctagcct ggtcactgtc
     5761 acaacgtcgc cagggcgtag gtggtcaagc atcctggcca gctccgggcg gtcgcgcctg
     5821 gtgccggtga tcttctcgga aaacagcttg gtgcagccgg ccgcgtgcag ttcggcccgt
     5881 tggttggtca agtcctggtc gtcggtgctg acgcgggcat agcccagcag gccagcggcg
     5941 gcgctcttgt tcatggcgta atgtctccgg ttctagtcgc aagtattcta ctttatgcga
     6001 ctaaaacacg cgacaagaaa acgccaggaa aagggcaggg cggcagcctg tcgcgtaact
     6061 taggacttgt gcgacatgtc gttttcagaa gacggctgca ctgaacgtca gaagccgact
     6121 gcactatagc agcggagggg ttggatcaaa gtactttaaa gtactttaaa gtactttaaa
     6181 gtactttgat cccgagggga accctgtggt tggcatgcac atacaaatgg acgaacggat
     6241 aaaccttttc acgccctttt aaatatccga ttattctaat aaacgctctt ttctcttagg
     6301 tttacccgcc aatatatcct gtcaaacact gatagtttaa actgaaggcg ggaaacgaca
     6361 atctgatcca agctcaagct aagcttgagc tctcccatat ggtcgactag agccaagctg
     6421 atctcctttg ccccggagat caccatggac gactttctct atctctacga tctaggaaga
     6481 aagttcgacg gagaaggtga cgataccatg ttcaccaccg ataatgagaa gattagcctc
     6541 ttcaatttca gaaagaatgc tgacccacag atggttagag aggcctacgc ggcaggtctc
     6601 atcaagacga tctacccgag taataatctc caggagatca aataccttcc caagaaggtt
     6661 aaagatgcag tcaaaagatt caggactaac tgcatcaaga acacagagaa agatatattt
     6721 ctcaagatca gaagtactat tccagtatgg acgattcaag gcttgcttca taaaccaagg
     6781 caagtaatag agattggagt ctctaagaaa gtagttccta ctgaatcaaa ggccatggag
     6841 tcaaaaattc agatcgagga tctaacagaa ctcgccgtga agactggcga acagttcata
     6901 cagagtcttt tacgactcaa tgacaagaag aaaatcttcg tcaacatggt ggagcacgac
     6961 actctcgtct actccaagaa tatcaaagat acagtctcag aagaccaaag ggctattgag
     7021 acttttcaac aaagggtaat atcgggaaac ctcctcggat tccattgccc agctatctgt
     7081 cacttcatca aaaggacagt agaaaaggaa ggtggcacct acaaatgcca tcattgcgat
     7141 aaaggaaagg ctatcgttca agatgcccct gccgacagtg gtcccaaaga tggaccccca
     7201 cccacgagga gcatcgtgga aaaagaagac gttccaacca cgtcttcaaa gcaagtggat
     7261 tgatgtgata tctccactga cgtaagggat gacgcacaat cccactatcc ttcgcaagac
     7321 ccttcctcta tataaggaag ttcatttcat ttggagagga ctccggtatt tttacaacaa
     7381 ttaccacaac aaaacaaaca acaaacaaca ttacaattta ctattctagt cgacctgcag
     7441 gcggccgcac tagtatggcc tcctccgagg acgtcatcaa ggagttcatg cgcttcaagg
     7501 tgcgcatgga gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc
     7561 gcccctacga gggcacccag accgccaagc tgaaggtgac caagggcggc cccctgccct
     7621 tcgcctggga catcctgtcc cctcagttcc agtacggctc caaggcctac gtgaagcacc
     7681 ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag tgggagcgcg
     7741 tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg
     7801 gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtaa
     7861 tgcagaagaa gaccatgggc tgggaggcct ccaccgagcg gatgtacccc gaggacggcg
     7921 ccctgaaggg cgagatcaag atgaggctga agctgaagga cggcggccac tacgacgccg
     7981 aggtcaagac cacctacatg gccaagaagc ccgtgcagct gcccggcgcc tacaagaccg
     8041 acatcaagct ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgagc
     8101 gcgccgaggg ccgccactcc accggcgccg atatcacaag tttgtacaaa aaagctgaac
     8161 gagaaacgta aaatgatata aatatcaata tattaaatta gattttgcat aaaaaacaga
     8221 ctacataata ctgtaaaaca caacatatcc agtcactatg gcggccgcat taggcacccc
     8281 aggctttaca ctttatgctt ccggctcgta taatgtgtgg attttgagtt aggatccgtc
     8341 gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat ataccaccgt
     8401 tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg
     8461 tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg taaagaaaaa
     8521 taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga atgctcatcc
     8581 ggaattccgt atggcaatga aagacggtga gctggtgata tgggatagtg ttcacccttg
     8641 ttacaccgtt ttccatgagc aaactgaaac gttttcatcg ctctggagtg aataccacga
     8701 cgatttccgg cagtttctac acatatattc gcaagatgtg gcgtgttacg gtgaaaacct
     8761 ggcctatttc cctaaagggt ttattgagaa tatgtttttc gtctcagcca atccctgggt
     8821 gagtttcacc agttttgatt taaacgtggc caatatggac aacttcttcg cccccgtttt
     8881 caccatgggc aaatattata cgcaaggcga caaggtgctg atgccgctgg cgattcaggt
     8941 tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg cttaatgaat tacaacagta
     9001 ctgcgatgag tggcagggcg gggcgtaaac gcgtggatcc ggcttactaa aagccagata
     9061 acagtatgcg tatttgcgcg ctgatttttg cggtataaga atatatactg atatgtatac
     9121 ccgaagtatg tcaaaaagag gtatgctatg aagcagcgta ttacagtgac agttgacagc
     9181 gacagctatc agttgctcaa ggcatatatg atgtcaatat ctccggtctg gtaagcacaa
     9241 ccatgcagaa tgaagcccgt cgtctgcgtg ccgaacgctg gaaagcggaa aatcaggaag
     9301 ggatggctga ggtcgcccgg tttattgaaa tgaacggctc ttttgctgac gagaacaggg
     9361 gctggtgaaa tgcagtttaa ggtttacacc tataaaagag agagccgtta tcgtctgttt
     9421 gtggatgtac agagtgatat tattgacacg cccgggcgac ggatggtgat ccccctggcc
     9481 agtgcacgtc tgctgtcaga taaagtctcc cgtgaacttt acccggtggt gcatatcggg
     9541 gatgaaagct ggcgcatgat gaccaccgat atggccagtg tgccggtctc cgttatcggg
     9601 gaagaagtgg ctgatctcag ccaccgcgaa aatgacatca aaaacgccat taacctgatg
     9661 ttctggggaa tataaatgtc aggctccctt atacacagcc agtctgcagg tcgaccatag
     9721 tgactggata tgttgtgttt tacagtatta tgtagtctgt tttttatgca aaatctaatt
     9781 taatatattg atatttatat cattttacgt ttctcgttca gctttcttgt acaaagtggt
     9841 gatatcccgc ggccatgcta gagtccgcaa aaatcaccag tctctctcta caaatctatc
     9901 tctctctatt tttctccaga ataatgtgtg agtagttccc agataaggga attagggttc
     9961 ttatagggtt tcgctcatgt gttgagcata taagaaaccc ttagtatgta tttgtatttg
    10021 taaaatactt ctatcaataa aatttctaat tcctaaaacc aaaatccagt gacctgcagg
    10081 catgcgacgt cgggccctct agaggatccc cgggtaccgc gaattatcga tcatgagcgg
    10141 agaattaagg gagtcacgtt atgacccccg ccgatgacgc gggacaagcc gttttacgtt
    10201 tggaactgac agaaccgcaa cgttgaagga gccactgagc cgcgggtttc tggagtttaa
    10261 tgagctaagc acatacgtca gaaaccatta ttgcgcgttc aaaagtcgcc taaggtcact
    10321 atcagctagc aaatatttct tgtcaaaaat gctccactga cgttccataa attcccctcg
    10381 gtatccaatt agagtctcat attcactctc aactcgatcg aggggatcta ccatgagccc
    10441 agaacgacgc ccggccgaca tccgccgtgc caccgaggcg gacatgccgg cggtctgcac
    10501 catcgtcaac cactacatcg agacaagcac ggtcaacttc cgtaccgagc cgcaggaacc
    10561 gcaggagtgg acggacgacc tcgtccgtct gcgggagcgc tatccctggc tcgtcgccga
    10621 ggtggacggc gaggtcgccg gcatcgccta cgcgggtccc tggaaggcac gcaacgccta
    10681 cgactggacg gccgagtcga ccgtgtacgt ctccccccgc caccagcgga cgggactggg
    10741 ctccacgctc tacacccacc tgctgaagtc cctggaggca cagggcttca agagcgtggt
    10801 cgctgtcatc gggctgccca acgacccgag cgtgcgcatg cacgaggcgc tcggatatgc
    10861 cccccgcggc atgctgcggg cggccggctt caagcacggg aactggcatg acgtgggttt
    10921 ctggcagctg gacttcagcc tgccggtgcc gccccgtccg gtcctgcccg tcaccgaaat
    10981 ctgatgaccc ctagagtcaa gcagatcgtt caaacatttg gcaataaagt ttcttaagat
    11041 tgaatcctgt tgccggtctt gcgatgatta tcatataatt tctgttgaat tacgttaagc
    11101 atgtaataat taacatgtaa tgcatgacgt tatttatgag atgggttttt atgattagag
    11161 tcccgcaatt atacatttaa tacgcgatag aaaacaaaat atagcgcgca aactaggata
    11221 aattatcgcg cgcggtgtca tctatgttac tagatcgacc ggcatgcaag ctgataattc
    11281 aattcggcgt taattcagta cattaaaaac gtccgcaatg tgttattaag ttgtctaagc
    11341 gtcaatttgt ttacaccaca atatatcctg ccaccagcca gccaacagct ccccgaccgg
    11401 cagctcggca caaaatcacc actcgataca ggcagcccat cagtccggga cggcgtcagc
    11461 gggagagccg ttgtaaggcg gcagactttg ctcatgttac cgatgctatt cggaagaacg
    11521 gcaactaagc tgccgggttt gaaacacgga tgatctcgcg gagggtagca tgttgattgt
    11581 aacgatgaca gagcgttgct gcctgtgatc a
//
