LOCUS       pPm43GW                 8560 bp    DNA     circular     07-JUL-2022
DEFINITION  .
ACCESSION   urn.local...lv-doz2q04
KEYWORDS    .
SOURCE      
  ORGANISM  .
COMMENT     http://www.informaxinc.com/                                        
            This file is created by Vector NTI                                 
            http://www.invitrogen.com/                                         
            ORIGDB|GenBank                                                     
            VNTDATE|342100386|                                                 
            VNTDBDATE|457776778|                                               
            LSOWNER|                                                           
            VNTNAME|pPm43GW,0|                                                 
            VNTAUTHORNAME|Demo User|                                           
            VNTAUTHOREML|manka@psb.ugent.be|                                   
            VNTOAUTHORNAME|Mansour Karimi|                                     
            VNTOAUTHORTEL|+3193313850|                                         
            VNTOAUTHOREML|mansour.karimi@psb.ugent.be|                         
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                                       #243#)) (CObjectList))) (CObjectList))  
                     #245=(CGroupWidget (CWidget 11 (8 0) 1 2 0 0 Nil -1996
            100)                  (CObjectList                                 
                                   #246=(CGroupWidget                          
                                         (CWidget 1 (10 #1# 0) 1 2 0 0 Nil -430
            100)                         (CObjectList                          
                                          #247=(CScratch                       
                                                (CWidget 1 (1 #1# 1) 1 2 0 0
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                                                                   (CWidget 0
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                          "@N (@S)" 5 2.23703 16.213 0 -10 2.15833             
                              0.694805 #247#)) (CObjectList))                  
                 #249=(CGroupWidget                                            
                       (CWidget 1 (10 #5# 0) 1 2 0 0 Nil -900 100)             
                       (CObjectList                                            
                        #250=(CScratch                                         
                              (CWidget 1 (1 #5# 1) 1 2 0 0 #166# 18023072      
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                                      0.123113 1)                              
                                #251=(CLabel                                   
                                      (CWidget 0 (0 0) 1 2 0 0 #195# 14398892
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                                                (LOGFONT 47 17 0 0 400 255 1 0
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            0.611111 0                                        "{\\i Cla}I
            (5067)" "@N (@S)" 5 -9.83473                               
            -10.4226 0 -10 2.9714 0.694805 #250#))                        
            (CObjectList))                                               
            #252=(CGroupWidget                                                 
                  (CWidget 1 (10 #8# 0) 1 2 0 0 Nil -351 100)                  
                  (CObjectList                                                 
                   #253=(CScratch                                              
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            14.8713 0                                0 -10 3.6662 0.694805
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                                                           (CWidget 1 (10 #9#
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              #257=(CLabel                                                     
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                                 "{\\i Nco}I (1010)" "@N (@S)" 5 13.4678       
                                    5.79035 0 -10 3.11923 0.694805 #256#))     
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                         #258=(CGroupWidget                                    
                               (CWidget 1 (10 #13# 0) 1 2 0 0 Nil -821 100)    
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                                 #259=(CScratch                                
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            #261=(CGroupWidget                                                 
                  (CWidget 1 (10 #14# 0) 1 2 0 0 Nil -1213 100)                
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            (1811)" "@N (@S)" 5 14.5568                                1.15807
            0 -10 3.07488 0.694805 #262#))                        
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            #264=(CGroupWidget                                                 
                  (CWidget 1 (10 #16# 0) 1 2 0 0 Nil -1918 100)                
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                     #265=(CScratch                                            
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            (183)" "@N (@S)" 5 4.48881                                12.7388 0
            -10 2.61661 0.694805 #265#))                         (CObjectList))
                                                           #267=(CGroupWidget  
                                                                 (CWidget 1 (10
            #19# 0) 1 2 0 0 Nil -510 100)                         (CObjectList 
                                                                  
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            14.5665                                -2.31614 0 -10 3.11923
            0.694805 #268#))                         (CObjectList))            
                                               #270=(CGroupWidget              
                                                     (CWidget 2 (10 #6# 0) 1 2
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            1) 1 2 0 0 #166# 96266844                                 100)
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            #272=(CLabel                                                       
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                               "{\\i Eco}RI (1311)" "@N (@S)" 5 14.1067        
                                   4.63228 0 -10 3.50359 0.694805 #271#)       
                              #273=(CScratch                                   
                                    (CWidget 2 (1 #6# 2) 1 2 0 0 #166# 18022816
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            (8508)" "@N (@S)" 5 -8.331                                11.5807 0
            -10 3.50359 0.694805 #273#))                         (CObjectList))
                                                           #275=(CGroupWidget  
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            #10# 0) 1 2 0 0 Nil -430 100)                         (CObjectList 
                                                                  
            #276=(CScratch                                                     
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                                              "{\\i Nde}I (3532)" "@N (@S)" 5
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                                   -3.47421 0 -10 3.16358 0.694805 #278#))     
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                         #280=(CGroupWidget                                    
                               (CWidget 2 (10 #17# 0) 1 2 0 0 Nil -116 100)    
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                                 #281=(CScratch                                
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            "{\\i Sma}I (444)" "@N (@S)" 5 11.1046                             
              9.26456 0 -10 2.98618 0.694805 #281#)                         
            #283=(CScratch                                                     
                  (CWidget 2 (1 #17# 2) 1 2 0 0 #166# 40607288                 
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                                          "{\\i Sma}I (8523)" "@N (@S)" 5
            -2.73241                                13.8968 0 -10 3.29663
            0.694805 #283#))                         (CObjectList))            
                                               #285=(CGroupWidget              
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            #20# 1) 1 2 0 0 #166# 18023168                                 100)
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                               "{\\i Xba}I (864)" "@N (@S)" 5 11.976 8.10649   
                                        0 -10 2.85313 0.694805 #286#)          
                                  #288=(CScratch                               
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            "{\\i Xba}I (8532)" "@N (@S)" 5 -2.62868                           
                15.0549 0 -10 3.16358 0.694805 #288#))                        
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            #290=(CGroupWidget                                                 
                  (CWidget 2 (10 #22# 0) 1 2 0 0 Nil -980 100)                 
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                    #291=(CScratch                                             
                          (CWidget 1 (1 #22# 1) 1 2 0 0 #166# 1 100)           
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                                 #292=(CLabel (CWidget 0 (0 0) 1 2 0 0 #194# 0
            100)                                (LOGPEN 0 0 8437824) 1         
                                                (LOGFONT 47 17 0 0 400 255 0 0
            0 3 2 1 34                                 "Arial") 1.15807
            0.611111 0                                        "{\\i Xma}I
            (442)" "@N (@S)" 5 9.83473                                10.4226 0
            -10 2.98618 0.694805 #291#)                          #293=(CScratch
                                                                       (CWidget
            2 (1 #22# 2) 1 2 0 0 #166# 18023176                                
            100) (LOGPEN 0 10 0) 8 1 0.0154241 1.9                             
              0.123113 1)                                                  
            #294=(CLabel                                                       
                  (CWidget 0 (0 0) 1 2 0 0 #194# 41014244 100)                 
                          (LOGPEN 0 0 8437824) 1                               
                          (LOGFONT 47 17 0 0 400 255 0 0 0 3 2 1 34            
                                "Arial") 1.15807 0.611111 0                    
                               "{\\i Xma}I (8521)" "@N (@S)" 5 -4.83044        
                                   12.7388 0 -10 3.29663 0.694805 #293#))      
                              (CObjectList))) (CObjectList))                   
                 #295=(CGroupWidget (CWidget 14 (16 0) 1 2 0 0 Nil -275 100)   
                          (CObjectList) (CObjectList))                         
                    #296=(CGroupWidget (CWidget 12 (0 0) 1 2 0 0 Nil -1056 100)
                             (CObjectList) (CObjectList))) (CObjectList)))     
               (CSeqView 10 5 (CObjectList) (CObList) 1 (CObList)) (CObList) 1 
               (CStringList) 1 36854592 (CObList))).
FEATURES             Location/Qualifiers
     T-DNA_border    1..333
                     /modified_by="wacae"
                     /note="Derived using Geneious Prime 2020.2.2 '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     70..89
                     /%GC="50.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="59.9"
                     /annotation_group="598665335042330040b41dd4-5ba3-4c40-8e48
                     -502e56cbdd57"
                     /created_by="primer3"
                     /Sequence="TCAAACCCGGCAGCTTAGTT"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pPm43GW"
                     /label="P8"
     misc_feature    336^337
                     /Original_Bases="GAATT"
                     /label="GAATT"
                     /note="Geneious type: Editing History Deletion"
     misc_feature    339
                     /Original_Bases="A"
                     /label="A"
                     /note="Geneious type: Editing History Replacement"
     misc_feature    341^342
                     /Original_Bases="G"
                     /label="G"
                     /note="Geneious type: Editing History Deletion"
     primer          387..403
                     /label="M13 R"
     attR4           423..546
                     /created_by="mamee"
                     /note="Derived using Geneious Prime 2022.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="attR4"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; attR4'
                     (<a href=""urn:local:.:l1-eztyg66"">attR4</a>)"
                     /Alternative_Match="99.19%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:o4-f0pbp83"">attR4</a>"
                     /Color="#00cccc"
                     /modified_by="mamee"
     CDS             complement(587..892)
                     /note="Derived using Geneious Prime 2020.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="ccdB"
                     /Transferred_Similarity="99.35%"
                     /Primary_Match="99.35%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:8r-dgycryy"">ccdB</a>"
                     /label="ccdB"
     primer_bind     847..867
                     /%GC="47.6"
                     /Hairpin_Tm="45.0"
                     /Self_Dimer_Tm="None"
                     /Tm="57.6"
                     /annotation_group="5986653583251700bff09e1f-261a-40c0-9714
                     -093e58de5791"
                     /created_by="primer3"
                     /Sequence="GACGATAACGGCTCTCTCTTT"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pPm43GW"
                     /label="P15"
     UTR             complement(893..1211)
                     /Mismatches=1
                     /%_Identity="99.68652037617555"
                     /Motif="AAGCCAGATAACAGTATGCGTATTTGCGCGCTGATTTTTGCGGTATAAGA
                     ATATATACTGATATGTATACCCGAAGTATGTCAAAAGGAGGTATGCTATGAAGCAGCG
                     TATTACAGTGACAGTTGACAGCGACAGCTATCAGTTGCTCAAGGCATATATGATGTCA
                     ATATCTCCGGTCTGGTAAGCACAACCATGCAGAATGAAGCCCGTCGTCTGCGTGCCGA
                     ACGCTGGAAAGCGGAAAATCAGGAAGGGATGGCTGAGGTCGCCCGGTTTATTGAAATG
                     AACGGCTCTTTTGCTGACGAGAACAGGGGCTGGTGAA"
                     /note="Derived using Geneious Prime 2020.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="ccdB"
                     /Transferred_Similarity="98.43%"
                     /Primary_Match="98.43%; Adjusted-Interval-0:0/0; ccdB
                     5'UTR (<a href=""urn:local:.:8r-dgycryy"">ccdB</a>)"
                     /label="ccdB 5'UTR"
     misc_feature    966
                     /Original_Bases="C"
                     /label="C"
                     /note="Geneious type: Editing History Replacement"
     misc_feature    1045
                     /Original_Bases="T"
                     /label="T"
                     /note="Geneious type: Editing History Replacement"
     misc_feature    complement(1234..1892)
                     /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="99.85%"
                     /Primary_Match="99.85%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:29-f0o4v7s"">CmR</a>"
                     /label="CmR"
                     /note="Geneious type: Bacterial Selection"
     misc_feature    1245^1246
                     /Original_Bases="C"
                     /label="C"
                     /note="Geneious type: Editing History Deletion"
     primer_bind     1643..1662
                     /%GC="55.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="60.0"
                     /annotation_group="598665364176830076c6d5d1-c451-46e6-a5c3
                     -e6836c08b3da"
                     /created_by="primer3"
                     /Sequence="TCACCAGCTCACCGTCTTTC"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pPm43GW"
                     /label="P16"
     promoter        complement(1946..1976)
                     /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    1984^1985
                     /Original_Bases="G"
                     /label="G"
                     /note="Geneious type: Editing History Deletion"
     attR3           complement(2000..2124)
                     /created_by="mamee"
                     /Color="#00cccc"
                     /modified_by="mamee"
                     /note="Derived using Geneious Prime 2022.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="attR3"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:kw-eztyb8e"">attR3</a>"
     primer          2133..2148
                     /label="M13F"
     T-DNA_border    2176..2406
                     /modified_by="wacae"
                     /note="Derived using Geneious Prime 2020.2.2 '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>"
                     /Alternative_Match="99.50%; Adjusted-Interval-0:0/-30; <a
                     href=""urn:local:.:3c-djznfva"">RB</a>"
                     /label="RB"
     primer_bind     2197..2216
                     /%GC="50.0"
                     /Hairpin_Tm="36.2"
                     /Self_Dimer_Tm="None"
                     /Tm="60.0"
                     /annotation_group="59866535250757000f4c125f-ffd5-47a0-8058
                     -5992470f56b9"
                     /created_by="primer3"
                     /Sequence="AGATTGTCGTTTCCCGCCTT"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pPm43GW"
                     /label="P14"
     misc_feature    2291
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     misc_feature    2384..2413
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     primer_bind     3025..3043
                     /%GC="63.2"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="61.3"
                     /annotation_group="59866529385529008a8d1e07-ca8f-498f-b5ba
                     -e7df70239c8b"
                     /created_by="primer3"
                     /Sequence="CCCGAGGCGTGAAGTTTGG"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pPm43GW"
                     /label="P1"
     regulatory      3592..4221
                     /note="Pseudomonas pVS1 plasmid stability protein"
                     /product="plasmid stability protein StaA"
                     /gene="staA"
                     /regulatory_class="other"
                     /note="Derived using Geneious Prime 2020.2.2 '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     3817..3836
                     /%GC="55.0"
                     /Hairpin_Tm="42.2"
                     /Self_Dimer_Tm="None"
                     /Tm="60.0"
                     /annotation_group="598665300033310049ac3501-adbf-42dc-9b3a
                     -c5c1d58490f1"
                     /created_by="primer3"
                     /Sequence="CGCGACTTCGTAGTGATCGA"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pPm43GW"
                     /label="P2"
     primer_bind     4614..4632
                     /%GC="63.2"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="61.0"
                     /annotation_group="598665305988010082e6996b-9511-491f-9312
                     -dfd38918a94f"
                     /created_by="primer3"
                     /Sequence="CAATGGCACTGGAACCCCC"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pPm43GW"
                     /label="P3"
     regulatory      4650..5723
                     /note="Pseudomonas pVS1 plasmid replication protein"
                     /product="replication protein RepA"
                     /gene="repA"
                     /regulatory_class="other"
                     /note="Derived using Geneious Prime 2020.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="pVS1 repA"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:7b-dlucaql"">pVS1 repA</a>"
                     /label="pVS1 repA"
     misc_feature    4654
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     primer_bind     5418..5435
                     /%GC="61.1"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="0.7"
                     /Tm="60.0"
                     /annotation_group="5986653117710200d7d8d119-3da7-4d3f-a836
                     -44f24aa44655"
                     /created_by="primer3"
                     /Sequence="AAGAACCCGGACGTGCTG"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pPm43GW"
                     /label="P4"
     rep_origin      5789..5983
                     /note="origin of replication fromÂ Pseudomonas pVS1
                     plasmid"
                     /modified_by="mamee"
                     /note="Derived using Geneious Prime 2020.2.2 '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"
     primer_bind     6218..6237
                     /%GC="55.0"
                     /Hairpin_Tm="33.5"
                     /Self_Dimer_Tm="None"
                     /Tm="60.0"
                     /annotation_group="5986653176016200b26871d8-73dc-4e95-b5bd
                     -60740c0a30d6"
                     /created_by="primer3"
                     /Sequence="ACCTCTGACACATGCAGCTC"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pPm43GW"
                     /label="P5"
     regulatory      6357..6617
                     /%_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 2020.2.2 '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>"
                     /Alternative_Match="99.29%; Adjusted-Interval-0:-30/-150;
                     <a href=""urn:local:.:79-dlucaql"">bom</a>"
                     /label="pBR322 bom"
     rep_origin      complement(6653..7241)
                     /created_by="thjac"
                     /modified_by="mamee"
                     /note="Derived using Geneious Prime 2020.2.2 '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     7042..7061
                     /%GC="55.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="58.9"
                     /annotation_group="5986653233020400ae411f6a-2011-4ce7-9c28
                     -82f9b1444048"
                     /created_by="primer3"
                     /Sequence="GCGGTGCTACAGAGTTCTTG"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pPm43GW"
                     /label="P6"
     misc_feature    7427^7428
                     /Original_Bases="T"
                     /label="T"
                     /note="Geneious type: Editing History Deletion"
     misc_feature    7439^7440
                     /Original_Bases="C"
                     /label="C"
                     /note="Geneious type: Editing History Deletion"
     misc_feature    7444..7447
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     misc_feature    complement(7487..8278)
                     /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     7814..7833
                     /%GC="55.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="60.1"
                     /annotation_group="5986653290212000b8de6960-ecbf-4397-9224
                     -42e672764fae"
                     /created_by="primer3"
                     /Sequence="ATCAAAGAGTTCCTCCGCCG"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pPm43GW"
                     /label="P7"
     Promoter        complement(8279..8553)
                     /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    8413^8414
                     /Original_Bases="T"
                     /label="T"
                     /note="Geneious type: Editing History Deletion"
     misc_feature    8424^8425
                     /Original_Bases="AAG"
                     /label="AAG"
                     /note="Geneious type: Editing History Deletion"
     misc_feature    8492..8493
                     /Original_Bases="CG"
                     /label="CG"
                     /note="Geneious type: Editing History Replacement"
     misc_feature    8496^8497
                     /Original_Bases="GT"
                     /label="GT"
                     /note="Geneious type: Editing History Deletion"
     misc_feature    8529^8530
                     /Original_Bases="G"
                     /label="G"
                     /note="Geneious type: Editing History Deletion"
ORIGIN      
        1 tgatcacagg cagcaacgct ctgtcatcgt tacaatcaac atgctaccct ccgcgagatc
       61 atccgtgttt caaacccggc agcttagttg ccgttcttcc gaatagcatc ggtaacatga
      121 gcaaagtctg ccgccttaca acggctctcc cgctgacgcc gtcccggact gatgggctgc
      181 ctgtatcgag tggtgatttt gtgccgagct gccggtcggg gagctgttgg ctggctggtg
      241 gcaggatata ttgtggtgta aacaaattga cgcttagaca acttaataac acattgcgga
      301 cgtttttaat gtactgaatt aacgccgaat tgaattcgcg gtacccgggg atcctctaga
      361 gggcccgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct
      421 atcaactttg tatagaaaag ttgaacgaga aacgtaaaat gatataaata tcaatatatt
      481 aaattagatt ttgcataaaa aacagactac ataatactgt aaaacacaac atatccagtc
      541 actatggtcg acctgcagac tggctgtgta taagggagcc tgacatttat attccccaga
      601 acatcaggtt aatggcgttt ttgatgtcat tttcgcggtg gctgagatca gccacttctt
      661 ccccgataac ggagaccggc acactggcca tatcggtggt catcatgcgc cagctttcat
      721 ccccgatatg caccaccggg taaagttcac gggggacttt atctgacagc agacgtgcac
      781 tggccagggg gatcaccatc cgtcgcccgg gcgtgtcaat aatatcactc tgtacatcca
      841 caaacagacg ataacggctc tctcttttat aggtgtaaac cttaaactgc atttcaccag
      901 cccctgttct cgtcggcaaa agagccgttc atttcaataa accgggcgac ctcagccatc
      961 ccttcttgat tttccgcttt ccagcgttcg gcacgcagac gacgggcttc attctgcatg
     1021 gttgtgctta ccgaaccgga gatactgaca tcatatatgc cttgagcaac tgatagctgt
     1081 cgctgtcaac tgtcactgta atacgctgct tcatagcata cctctttttg acatacttcg
     1141 ggtatacata tcagtatata ttcttatacc gcaaaaatca gcgcgcaaat acgcatactg
     1201 ttatctggct tttagtaagc cggatcctct agattacgcc ccgcctgcca ctcatcgcag
     1261 tactgttgta attcattaag cattctgccg acatggaagc catcacaaac ggcatgatga
     1321 acctgaatcg ccagcggcat cagcaccttg tcgccttgcg tataatattt gcccatggtg
     1381 aaaacggggg cgaagaagtt gtccatattg gccacgttta aatcaaaact ggtgaaactc
     1441 acccagggat tggctgagac gaaaaacata ttctcaataa accctttagg gaaataggcc
     1501 aggttttcac cgtaacacgc cacatcttgc gaatatatgt gtagaaactg ccggaaatcg
     1561 tcgtggtatt cactccagag cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa
     1621 caagggtgaa cactatccca tatcaccagc tcaccgtctt tcattgccat acggaattcc
     1681 ggatgagcat tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa cttgtgctta
     1741 tttttcttta cggtctttaa aaaggccgta atatccagct gaacggtctg gttataggta
     1801 cattgagcaa ctgactgaaa tgcctcaaaa tgttctttac gatgccattg ggatatatca
     1861 acggtggtat atccagtgat ttttttctcc attttagctt ccttagctcc tgaaaatctc
     1921 gacggatcct aactcaaaat ccacacatta tacgagccgg aagcataaag tgtaaagcct
     1981 ggggtgccta atgcggccgc catagtgact ggatatgttg tgttttacag tattatgtag
     2041 tctgtttttt atgcaaaatc taatttaata tattgatatt tatatcattt tacgtttctc
     2101 gttcaacttt attatacata gttgataatt cactggccgt cgttttacaa cgtcgtgact
     2161 gggaaaacga gctcaagctt agcttgagct tggatcagat tgtcgtttcc cgccttcagt
     2221 ttaaactatc agtgtttgac aggatatatt ggcgggtaaa cctaagagaa aagagcgttt
     2281 attagaataa tcggatattt aaaagggcgt gaaaaggttt atccgttcgt ccatttgtat
     2341 gtgcatgcca accacagggt tcccctcggg atcaaagtac tttaaagtac tttaaagtac
     2401 tttaaagtac tttgatccaa cccctccgct gctatagtgc agtcggcttc tgacgttcag
     2461 tgcagccgtc ttctgaaaac gacatgtcgc acaagtccta agttacgcga caggctgccg
     2521 ccctgccctt ttcctggcgt tttcttgtcg cgtgttttag tcgcataaag tagaatactt
     2581 gcgactagaa ccggagacat tacgccatga acaagagcgc cgccgctggc ctgctgggct
     2641 atgcccgcgt cagcaccgac gaccaggact tgaccaacca acgggccgaa ctgcacgcgg
     2701 ccggctgcac caagctgttt tccgagaaga tcaccggcac caggcgcgac cgcccggagc
     2761 tggccaggat gcttgaccac ctacgccctg gcgacgttgt gacagtgacc aggctagacc
     2821 gcctggcccg cagcacccgc gacctactgg acattgccga gcgcatccag gaggccggcg
     2881 cgggcctgcg tagcctggca gagccgtggg ccgacaccac cacgccggcc ggccgcatgg
     2941 tgttgaccgt gttcgccggc attgccgagt tcgagcgttc cctaatcatc gaccgcaccc
     3001 ggagcgggcg cgaggccgcc aaggcccgag gcgtgaagtt tggcccccgc cctaccctca
     3061 ccccggcaca gatcgcgcac gcccgcgagc tgatcgacca ggaaggccgc accgtgaaag
     3121 aggcggctgc actgcttggc gtgcatcgct cgaccctgta ccgcgcactt gagcgcagcg
     3181 aggaagtgac gcccaccgag gccaggcggc gcggtgcctt ccgtgaggac gcattgaccg
     3241 aggccgacgc cctggcggcc gccgagaatg aacgccaaga ggaacaagca tgaaaccgca
     3301 ccaggacggc caggacgaac cgtttttcat taccgaagag atcgaggcgg agatgatcgc
     3361 ggccgggtac gtgttcgagc cgcccgcgca cgtctcaacc gtgcggctgc atgaaatcct
     3421 ggccggtttg tctgatgcca agctggcggc ctggccggcc agcttggccg ctgaagaaac
     3481 cgagcgccgc cgtctaaaaa ggtgatgtgt atttgagtaa aacagcttgc gtcatgcggt
     3541 cgctgcgtat atgatgcgat gagtaaataa acaaatacgc aaggggaacg catgaaggtt
     3601 atcgctgtac ttaaccagaa aggcgggtca ggcaagacga ccatcgcaac ccatctagcc
     3661 cgcgccctgc aactcgccgg ggccgatgtt ctgttagtcg attccgatcc ccagggcagt
     3721 gcccgcgatt gggcggccgt gcgggaagat caaccgctaa ccgttgtcgg catcgaccgc
     3781 ccgacgattg accgcgacgt gaaggccatc ggccggcgcg acttcgtagt gatcgacgga
     3841 gcgccccagg cggcggactt ggctgtgtcc gcgatcaagg cagccgactt cgtgctgatt
     3901 ccggtgcagc caagccctta cgacatatgg gccaccgccg acctggtgga gctggttaag
     3961 cagcgcattg aggtcacgga tggaaggcta caagcggcct ttgtcgtgtc gcgggcgatc
     4021 aaaggcacgc gcatcggcgg tgaggttgcc gaggcgctgg ccgggtacga gctgcccatt
     4081 cttgagtccc gtatcacgca gcgcgtgagc tacccaggca ctgccgccgc cggcacaacc
     4141 gttcttgaat cagaacccga gggcgacgct gcccgcgagg tccaggcgct ggccgctgaa
     4201 attaaatcaa aactcatttg agttaatgag gtaaagagaa aatgagcaaa agcacaaaca
     4261 cgctaagtgc cggccgtccg agcgcacgca gcagcaaggc tgcaacgttg gccagcctgg
     4321 cagacacgcc agccatgaag cgggtcaact ttcagttgcc ggcggaggat cacaccaagc
     4381 tgaagatgta cgcggtacgc caaggcaaga ccattaccga gctgctatct gaatacatcg
     4441 cgcagctacc agagtaaatg agcaaatgaa taaatgagta gatgaatttt agcggctaaa
     4501 ggaggcggca tggaaaatca agaacaacca ggcaccgacg ccgtggaatg ccccatgtgt
     4561 ggaggaacgg gcggttggcc aggcgtaagc ggctgggttg tctgccggcc ctgcaatggc
     4621 actggaaccc ccaagcccga ggaatcggcg tgagcggtcg caaaccatcc ggcccggtac
     4681 aaatcggcgc ggcgctgggt gatgacctgg tggagaagtt gaaggccgcg caggccgccc
     4741 agcggcaacg catcgaggca gaagcacgcc ccggtgaatc gtggcaagcg gccgctgatc
     4801 gaatccgcaa agaatcccgg caaccgccgg cagccggtgc gccgtcgatt aggaagccgc
     4861 ccaagggcga cgagcaacca gattttttcg ttccgatgct ctatgacgtg ggcacccgcg
     4921 atagtcgcag catcatggac gtggccgttt tccgtctgtc gaagcgtgac cgacgagctg
     4981 gcgaggtgat ccgctacgag cttccagacg ggcacgtaga ggtttccgca gggccggccg
     5041 gcatggccag tgtgtgggat tacgacctgg tactgatggc ggtttcccat ctaaccgaat
     5101 ccatgaaccg ataccgggaa gggaagggag acaagcccgg ccgcgtgttc cgtccacacg
     5161 ttgcggacgt actcaagttc tgccggcgag ccgatggcgg aaagcagaaa gacgacctgg
     5221 tagaaacctg cattcggtta aacaccacgc acgttgccat gcagcgtacg aagaaggcca
     5281 agaacggccg cctggtgacg gtatccgagg gtgaagcctt gattagccgc tacaagatcg
     5341 taaagagcga aaccgggcgg ccggagtaca tcgagatcga gctagctgat tggatgtacc
     5401 gcgagatcac agaaggcaag aacccggacg tgctgacggt tcaccccgat tactttttga
     5461 tcgatcccgg catcggccgt tttctctacc gcctggcacg ccgcgccgca ggcaaggcag
     5521 aagccagatg gttgttcaag acgatctacg aacgcagtgg cagcgccgga gagttcaaga
     5581 agttctgttt caccgtgcgc aagctgatcg ggtcaaatga cctgccggag tacgatttga
     5641 aggaggaggc ggggcaggct ggcccgatcc tagtcatgcg ctaccgcaac ctgatcgagg
     5701 gcgaagcatc cgccggttcc taatgtacgg agcagatgct agggcaaatt gccctagcag
     5761 gggaaaaagg tcgaaaaggt ctctttcctg tggatagcac gtacattggg aacccaaagc
     5821 cgtacattgg gaaccggaac ccgtacattg ggaacccaaa gccgtacatt gggaaccggt
     5881 cacacatgta agtgactgat ataaaagaga aaaaaggcga tttttccgcc taaaactctt
     5941 taaaacttat taaaactctt aaaacccgcc tggcctgtgc ataactgtct ggccagcgca
     6001 cagccgaaga gctgcaaaaa gcgcctaccc ttcggtcgct gcgctcccta cgccccgccg
     6061 cttcgcgtcg gcctatcgcg gccgctggcc gctcaaaaat ggctggccta cggccaggca
     6121 atctaccagg gcgcggacaa gccgcgccgt cgccactcga ccgccggcgc ccacatcaag
     6181 gcaccctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     6241 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     6301 tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga
     6361 gtgtatactg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
     6421 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt
     6481 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     6541 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     6601 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     6661 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     6721 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     6781 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     6841 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     6901 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     6961 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     7021 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     7081 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     7141 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     7201 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     7261 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgcatg
     7321 atatatctcc caatttgtgt agggcttatt atgcacgctt aaaaataata aaagcagact
     7381 tgacctgata gtttggctgt gagcaattat gtgcttagtg catctaacgc ttgagttaag
     7441 ccgcgccgcg aagcggcgtc ggcttgaacg aatttctagc tagacattat ttgccgacta
     7501 ccttggtgat ctcgcctttc acgtagtgga caaattcttc caactgatct gcgcgcgagg
     7561 ccaagcgatc ttcttcttgt ccaagataag cctgtctagc ttcaagtatg acgggctgat
     7621 actgggccgg caggcgctcc attgcccagt cggcagcgac atccttcggc gcgattttgc
     7681 cggttactgc gctgtaccaa atgcgggaca acgtaagcac tacatttcgc tcatcgccag
     7741 cccagtcggg cggcgagttc catagcgtta aggtttcatt tagcgcctca aatagatcct
     7801 gttcaggaac cggatcaaag agttcctccg ccgctggacc taccaaggca acgctatgtt
     7861 ctcttgcttt tgtcagcaag atagccagat caatgtcgat cgtggctggc tcgaagatac
     7921 ctgcaagaat gtcattgcgc tgccattctc caaattgcag ttcgcgctta gctggataac
     7981 gccacggaat gatgtcgtcg tgcacaacaa tggtgacttc tacagcgcgg agaatctcgc
     8041 tctctccagg ggaagccgaa gtttccaaaa ggtcgttgat caaagctcgc cgcgttgttt
     8101 catcaagcct tacggtcacc gtaaccagca aatcaatatc actgtgtggc ttcaggccgc
     8161 catccactgc ggagccgtac aaatgtacgg ccagcaacgt cggttcgaga tggcgctcga
     8221 tgacgccaac tacctctgat agttgagtcg atacttcggc gatcaccgct tcccccatga
     8281 tgtttaactt tgttttaggg cgactgccct gctgcgtaac atcgttgctg ctccataaca
     8341 tcaaacatcg acccacggcg taacgcgctt gctgcttgga tgcccgaggc atagactgta
     8401 ccccaaaaaa acagtcataa caagccatga aaaccgccac tgcgccgtta ccaccgctgc
     8461 gttcggtcaa ggttctggac cagttgcgtg agcgcatacg ctacttgcat tacagcttac
     8521 gaaccgaaca ggcttatgtc cactgggttc gtgcccgaat
//
