LOCUS       pEN-R2-8-L3             3409 bp    DNA     circular     06-SEP-2022
DEFINITION  .
ACCESSION   urn.local...132-dt4s1xu
KEYWORDS    .
SOURCE      
  ORGANISM  .
COMMENT     This file is created by Vector NTI                                 
            http://www.invitrogen.com/                                         
            ORIGDB|GenBank                                                     
            VNTDATE|459272238|                                                 
            VNTDBDATE|459272238|                                               
            LSOWNER|                                                           
            VNTNAME|pEN-R2-8-L3,0|                                             
            VNTAUTHORNAME|Demo User|                                           
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                                 (LOGPEN 0 0 8437824) 1                        
                                 (LOGFONT 43 16 0 0 400 255 0 0 0 3 2 1 34     
                                       "Arial") 1.15807 0.611111 0             
                                      "{\\i Apa}I (3371)" "@N (@S)" 5 -8.00674 
                                          11.5807 0 -10 3.0094 0.68832 #236#)) 
                                   (CObjectList))                              
                             #238=(CGroupWidget                                
                                   (CWidget 2 (10 #3# 0) 1 2 0 0 Nil -1488 100)
                                    (CObjectList                               
                                     #239=(CScratch                            
                                           (CWidget 1 (1 #3# 1) 1 2 0 0 #189# 0
            100)                                (LOGPEN 0 0 10079283) 1 1
            1.9257 1.9 0.123113                                1)              
                                                         #240=(CLabel (CWidget
            0 (0 0) 1 2 0 0 #168# 0 100)                                (LOGPEN
            0 0 8437824) 1                                             
            (LOGFONT 43 16 0 0 400 255 0 0 0 3 2 1 34                          
                  "Arial") 1.15807 0.611111 0                                  
                 "{\\i Ava}I (2446)" "@N (@S)" 5 -13.9876                      
                     -6.94842 0 -10 2.96138 0.68832 #239#)                     
                #241=(CScratch                                                 
                      (CWidget 2 (1 #3# 2) 1 2 0 0 #189# 0 100)                
                           (LOGPEN 0 0 10079283) 1 1 0.34851 1.9               
                            0.123113 1)                                        
                      #242=(CLabel (CWidget 0 (0 0) 1 2 0 0 #168# 0 100)       
                                    (LOGPEN 0 0 8437824) 1                     
                                    (LOGFONT 43 16 0 0 400 255 0 0 0 3 2 1 34  
                                          "Arial") 1.15807 0.611111 0          
                                         "{\\i Ava}I (3333)" "@N (@S)" 5
            -9.80578                                10.4226 0 -10 2.96138
            0.68832 #241#))                         (CObjectList))             
                                              #243=(CGroupWidget               
                                                    (CWidget 2 (10 #10# 0) 1 2
            0 0 Nil -186 100)                         (CObjectList             
                                                       #244=(CScratch          
                                                             (CWidget 1 (1 #10#
            1) 1 2 0 0 #189# 0 100)                                (LOGPEN 0 0
            10079283) 1 1 1.79056 1.9                                0.123113
            1)                                                   #245=(CLabel  
                                                                       (CWidget
            0 (0 0) 1 2 0 0 #168# 29769788 100)                               
            (LOGPEN 0 0 8437824) 1                                             
            (LOGFONT 43 16 0 0 400 255 0 0 0 3 2 1 34                          
                  "Arial") 1.15807 0.611111 0                                  
                 "{\\i Nde}I (2522)" "@N (@S)" 5 -14.5123 0 0                  
                         -10 3.0094 0.68832 #244#)                             
                   #246=(CScratch                                              
                         (CWidget 2 (1 #10# 2) 1 2 0 0 #189# 0 100)            
                               (LOGPEN 0 0 10079283) 1 1 1.57363 1.9           
                                0.123113 1)                                    
                          #247=(CLabel                                         
                                (CWidget 0 (0 0) 1 2 0 0 #168# 1944788432 100) 
                                          (LOGPEN 0 0 8437824) 1               
                                          (LOGFONT 43 16 0 0 400 255 0 0 0 3 2
            1 34                                 "Arial") 1.15807 0.611111 0   
                                                "{\\i Nde}I (2644)" "@N (@S)" 5
            -14.5218                                1.15807 0 -10 3.0094
            0.68832 #246#))                         (CObjectList))             
                                              #248=(CGroupWidget               
                                                    (CWidget 2 (10 #13# 0) 1 2
            0 0 Nil -331 100)                         (CObjectList             
                                                       #249=(CScratch          
                                                             (CWidget 1 (1 #13#
            1) 1 2 0 0 #189# 0 100)                                (LOGPEN 0 0
            10079283) 1 1 1.90436 1.9                                0.123113
            1)                                                   #250=(CLabel
            (CWidget 0 (0 0) 1 2 0 0 #168# 0 100)                              
             (LOGPEN 0 0 8437824) 1                                            
             (LOGFONT 43 16 0 0 400 255 0 0 0 3 2 1 34                         
                   "Arial") 1.15807 0.611111 0                                 
                  "{\\i Pst}I (2458)" "@N (@S)" 5 -13.8439                     
                      -4.63228 0 -10 2.8013 0.68832 #249#)                     
                #251=(CScratch                                                 
                      (CWidget 2 (1 #13# 2) 1 2 0 0 #189# 0 100)               
                            (LOGPEN 0 0 10079283) 1 1 0.103131 1.9             
                              0.123113 1)                                      
                        #252=(CLabel (CWidget 0 (0 0) 1 2 0 0 #168# 0 100)     
                                      (LOGPEN 0 0 8437824) 1                   
                                      (LOGFONT 43 16 0 0 400 255 0 0 0 3 2 1 34
                                            "Arial") 1.15807 0.611111 0        
                                           "{\\i Pst}I (3471)" "@N (@S)" 5
            -4.55829                                12.7388 0 -10 2.8013
            0.68832 #251#))                         (CObjectList))             
                                              #253=(CGroupWidget               
                                                    (CWidget 2 (10 #14# 0) 1 2
            0 0 Nil -259 100)                         (CObjectList             
                                                       #254=(CScratch          
                                                             (CWidget 1 (1 #14#
            1) 1 2 0 0 #189# 0 100)                                (LOGPEN 0 0
            10079283) 1 1 6.10426 1.9                                0.123113
            1)                                                   #255=(CLabel
            (CWidget 0 (0 0) 1 2 0 0 #168# 0 100)                              
             (LOGPEN 0 0 8437824) 1                                            
             (LOGFONT 43 16 0 0 400 255 0 0 0 3 2 1 34                         
                   "Arial") 1.15807 0.611111 0                                 
                  "{\\i Sac}I (96)" "@N (@S)" 5 4.35339 13.8968                
                           0 -10 2.32108 0.68832 #254#)                        
                     #256=(CScratch                                            
                           (CWidget 2 (1 #14# 2) 1 2 0 0 #189# 0 100)          
                                 (LOGPEN 0 0 10079283) 1 1 1.8848 1.9 0.123113 
                                          1)                                   
                                    #257=(CLabel (CWidget 0 (0 0) 1 2 0 0 #168#
            0 100)                                (LOGPEN 0 0 8437824) 1       
                                                  (LOGFONT 43 16 0 0 400 255 0
            0 0 3 2 1 34                                 "Arial") 1.15807
            0.611111 0                                        "{\\i Sac}I
            (2469)" "@N (@S)" 5 -14.324                                -2.31614
            0 -10 2.96138 0.68832 #256#))                        
            (CObjectList))) (CObjectList))                        
            #258=(CGroupWidget (CWidget 14 (16 0) 1 2 0 0 Nil -116 100)        
                     (CObjectList) (CObjectList))                              
               #259=(CGroupWidget (CWidget 12 (0 0) 1 2 0 0 Nil -476 100)      
                       (CObjectList) (CObjectList))) (CObjectList)))        
            (CSeqView 10 5 (CObjectList) (CObList) 1 (CObList)) (CObList)
            50855959    (CStringList) 1078493184 327701 (CObList))).
FEATURES             Location/Qualifiers
     attR2           complement(1..126)
                     /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="attR2"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:kr-ezty4ze"">attR2</a>"
     misc_feature    1^2
                     /Original_Bases="CATAGTGACTGCATATGTTGTGTTTTACAGTATTATGTAGT
                     CTGTTTTTTATGCAAAATCTAATTTAATATATTGATATTTATATCATTTTACGTTTCT
                     CGTTCAACTTTCTTGTACAAAGT"
                     /label="CATAGTGACTGCATATGTTGTGTTTTACAGTATTATGTAGTCTGTTTTTT
                     ATGCAAAATCTAATTTAATATATTGATATTTATATCATTTTACGTTTCTCGTTCAACT
                     TTCTTGTACAAAGT"
                     /note="Geneious type: Editing History Deletion"
     misc_feature    126..127
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     terminator      128..894
                     /created_by="lapau"
                     /modified_by="mamee"
                     /note="Derived using Geneious Prime 2022.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="OCST"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; tOCS (<a
                     href=""urn:local:.:69-dgycryw"">OCST</a>)"
                     /label="tOCS"
     primer_bind     541..560
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-R2-8-L3"
                     /Sequence="CACCGAGCGGCGAACTAATA"
                     /Tm="60.2"
                     /%GC="55.0"
                     /Hairpin_Tm="44.6"
                     /Self_Dimer_Tm="13.0"
                     /modified_by="mamee"
                     /label="P57"
     misc_feature    895
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     attL3           complement(896..991)
                     /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="attL3"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:lo-eztz06f"">attL3</a>"
     primer          complement(1049..1069)
                     /note="Derived using Geneious Prime 2021.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="pCDB-Cas9 - GWM13R"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; GWM13R
                     (pCDB-Cas9 - GWM13R)"
                     /label="GWM13R"
     promoter        1130..1180
                     /created_by="chgai"
                     /X-Transferred_From="pCG034_pKan_Kan-CBE"
                     /X-Transferred_Similarity="100.00%"
                     /X-Primary_Match="100.00%; Adjusted-Interval-0:0/0;
                     Kan_promoter (<a
                     href=""urn:local:chgai:ll-ch8iv1b"">pCG034_pKan_Kan-CBE</a
                     >)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     Kan_promoter (<a
                     href=""urn:local:chgai:lo-ch8iv1b"">pCG040_ABE-Nuc-NLS_Kan
                     -reporter_control</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     Kan_promoter (<a
                     href=""urn:local:chgai:lr-ch8iv1b"">pCG043_Kan-ABE-Dest_co
                     ntrol_ccdB</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     Kan_promoter (<a
                     href=""urn:local:chgai:lu-ch8iv1c"">pCG046_ABE-LbCas12a-NL
                     S_Kan_control</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     Kan_promoter (<a
                     href=""urn:local:chgai:kx-ch8iv19"">pCG005_pGGD-pKan-KanR-
                     E</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     Kan_promoter (<a
                     href=""urn:local:chgai:ky-ch8iv19"">pCG006_pGGA-Kan_promot
                     er-B</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     Kan_promoter (<a
                     href=""urn:local:chgai:lm-ch8iv1b"">pCG035_pKan_Kan</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     Kan_promoter (<a
                     href=""urn:local:chgai:ln-ch8iv1b"">pCG039_ABE-N7-NLS_Kan-
                     reporter_control</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     Kan_promoter (<a
                     href=""urn:local:chgai:lp-ch8iv1b"">pCG041_ABE-SV40-NLS_Ka
                     n-reporter_control</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     Kan_promoter (<a
                     href=""urn:local:chgai:lw-ch8iv1c"">pCG054_Kan_CBE_control
                     </a>)"
                     /modified_by="mamee"
                     /note="Derived using Geneious Prime 2022.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="KanR"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; kan (<a
                     href=""urn:local:.:8j-dgycryy"">KanR</a>)"
                     /label="kan"
     misc_feature    1181..1990
                     /modified_by="wacae"
                     /note="Derived using Geneious Prime 2021.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="KanR"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:8j-dgycryy"">KanR</a>"
                     /label="KanR"
                     /note="Geneious type: Bacterial Selection"
     primer_bind     1206..1225
                     /%GC="55.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="25.9"
                     /Tm="59.8"
                     /annotation_group="6053150055719800c6079120-b214-42bf-8b94
                     -b9f980048ef6"
                     /created_by="primer3"
                     /Sequence="CGAGGCCGCGATTAAATTCC"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-R2-8-L3"
                     /label="P44"
     primer_bind     1840..1859
                     /%GC="60.0"
                     /Hairpin_Tm="40.0"
                     /Self_Dimer_Tm="None"
                     /Tm="60.0"
                     /annotation_group="6053149938814400ce5b4c6b-5072-419b-a932
                     -301187d7b92c"
                     /created_by="primer3"
                     /Sequence="CGCAGACCGATACCAGGATC"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-R2-8-L3"
                     /label="P42"
     rep_origin      2137..2725
                     /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="100.00%"
                     /Primary_Match="100.00%; 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"
     misc_feature    2284
                     /Original_Bases="C"
                     /label="C"
                     /note="Geneious type: Editing History Replacement"
     misc_feature    2529..2530
                     /Original_Bases="AT"
                     /label="AT"
                     /note="Geneious type: Editing History Replacement"
     primer_bind     2625..2644
                     /%GC="55.0"
                     /Hairpin_Tm="37.3"
                     /Self_Dimer_Tm="None"
                     /Tm="59.7"
                     /annotation_group="6053150118162700304d2321-1838-48d9-ac0b
                     -17444104f96e"
                     /created_by="primer3"
                     /Sequence="GGGGAAACGCCTGGTATCTT"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-R2-8-L3"
                     /label="P45"
     primer_bind     3233..3252
                     /%GC="55.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="60.0"
                     /annotation_group="60531501766424005c85fed7-f872-436c-a4d1
                     -1c7b760513f0"
                     /created_by="primer3"
                     /Sequence="CTTCCGACTGAGCCTTTCGT"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-R2-8-L3"
                     /label="P46"
     primer          3324..3344
                     /note="Derived using Geneious Prime 2021.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="pCDB-Cas9 - GWM13F"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; GWM13F
                     (pCDB-Cas9 - GWM13F)"
                     /label="GWM13F"
     misc_feature    3408
                     /Original_Bases="A"
                     /label="A"
                     /note="Geneious type: Editing History Replacement"
ORIGIN      
        1 tcatagtgac tgcatatgtt gtgttttaca gtattatgta gtctgttttt tatgcaaaat
       61 ctaatttaat atattgatat ttatatcatt ttacgtttct cgttcaactt tcttgtacaa
      121 agtgggactg ctttaatgag atatgcgaga cgcctatgat cgcatgatat ttgctttcaa
      181 ttctgttgtg cacgttgtaa aaaacctgag catgtgtagc tcagatcctt accgccggtt
      241 tcggttcatt ctaatgaata tatcacccgt tactatcgta tttttatgaa taatattctc
      301 cgttcaattt actgattgta ccctactact tatatgtaca atattaaaat gaaaacaata
      361 tattgtgctg aataggttta tagcgacatc tatgatagag cgccacaata acaaacaatt
      421 gcgttttatt attacaaatc caattttaaa aaaagcggca gaaccggtca aacctaaaag
      481 actgattaca taaatcttat tcaaatttca aaagtgcccc aggggctagt atctacgaca
      541 caccgagcgg cgaactaata acgctcactg aagggaactc cggttccccg ccggcgcgca
      601 tgggtgagat tccttgaagt tgagtattgg ccgtccgctc taccgaaagt tacgggcacc
      661 attcaacccg gtccagcacg gcggccgggt aaccgacttg ctgccccgag aattatgcag
      721 catttttttg gtgtatgtgg gccccaaatg aagtgcaggt caaaccttga cagtgacgac
      781 aaatcgttgg gcgggtccag ggcgaatttt gcgacaacat gtcgaggctc agcaggacct
      841 gcaggcatgc aagctagctt actagtgatg catattctat agtgtcacct aaatacaact
      901 ttattataca aagttggcat tataaaaaag cattgcttat caatttgttg caacgaacag
      961 gtcactatca gtcaaaataa aatcattatt tggagctcca tggtagcgtt aacgcggccg
     1021 cgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc
     1081 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat
     1141 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
     1201 aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     1261 tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc
     1321 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     1381 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     1441 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt
     1501 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     1561 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
     1621 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     1681 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
     1741 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     1801 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     1861 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     1921 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     1981 gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg
     2041 acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     2101 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     2161 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     2221 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     2281 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     2341 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     2401 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     2461 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     2521 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     2581 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     2641 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     2701 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     2761 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     2821 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     2881 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     2941 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     3001 gcgcaacgca attaatacgc gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg
     3061 ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct
     3121 gcccgccacc ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc
     3181 ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtcttccgac
     3241 tgagcctttc gttttatttg atgcctggca gttccctact ctcgcgttaa cgctagcatg
     3301 gatgttttcc cagtcacgac gttgtaaaac gacggccagt cttaagctcg ggccctgcag
     3361 ctctagagct cgaattctac aggtcactaa taccatctaa gtagttggt
//
