LOCUS       pEN-L4-2-R1             3769 bp    DNA     circular     06-SEP-2022
DEFINITION  .
ACCESSION   urn.local...r3-doze616
KEYWORDS    .
SOURCE      
  ORGANISM  .
COMMENT     This file is created by Vector NTI                                 
            http://www.invitrogen.com/                                         
            ORIGDB|GenBank                                                     
            VNTDATE|459275533|                                                 
            VNTDBDATE|459275533|                                               
            LSOWNER|                                                           
            VNTNAME|pEN-L4-2-R1,0|                                             
            VNTAUTHORNAME|Demo User|                                           
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            "Arial") 1.15807 0.611111 0                                       
            "{\\i Nde}I (156)" "@N (@S)" 5 5.58122                             
              12.7388 0 -10 2.68925 0.68832 #234#)                         
            #236=(CScratch                                                     
                  (CWidget 2 (1 #10# 2) 1 2 0 0 #186# 0 100)                   
                        (LOGPEN 0 0 10079283) 1 1 4.0654 1.9 0.123113          
                                 1)                                            
                           #237=(CLabel (CWidget 0 (0 0) 1 2 0 0 #165# 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 Nde}I (1325)" "@N (@S)" 5
            12.3746                                -9.26456 0 -10 3.0094
            0.68832 #236#))                         (CObjectList))             
                                              #238=(CGroupWidget               
                                                    (CWidget 2 (10 #11# 0) 1 2
            0 0 Nil -1477 100)                         (CObjectList            
                                                        #239=(CScratch         
                                                              (CWidget 1 (1
            #11# 1) 1 2 0 0 #186# 0 100)                                (LOGPEN
            0 0 10079283) 1 1 4.27884 1.9                               
            0.123113 1)                                                  
            #240=(CLabel (CWidget 0 (0 0) 1 2 0 0 #165# 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 Not}I (1197)" "@N (@S)" 5 13.4558         
                                  -5.79035 0 -10 2.84933 0.68832 #239#)        
                             #241=(CScratch                                    
                                   (CWidget 2 (1 #11# 2) 1 2 0 0 #186# 0 100)  
                                         (LOGPEN 0 0 10079283) 1 1 3.942 1.9
            0.123113                                1)                         
                                              #242=(CLabel (CWidget 0 (0 0) 1 2
            0 0 #165# 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 Not}I (1399)" "@N (@S)" 5 11.5229                            
               -12.7388 0 -10 2.84933 0.68832 #241#))                        
            (CObjectList))) (CObjectList))                        
            #243=(CGroupWidget (CWidget 14 (16 0) 1 2 0 0 Nil -932 100)        
                     (CObjectList) (CObjectList))                              
               #244=(CGroupWidget (CWidget 12 (0 0) 1 2 0 0 Nil -865 100)      
                       (CObjectList) (CObjectList))) (CObjectList)))        
            (CSeqView 10 5 (CObjectList) (CObList) 1 (CObList)) (CObList)
            40836432    (CStringList) 50856079 1610612736 (CObList))).
FEATURES             Location/Qualifiers
     attL4           1..97
                     /Color="#00cccc"
                     /created_by="mamee"
                     /note="Derived using Geneious Prime 2022.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="attL4"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:lr-eztz0t4"">attL4</a>"
     misc_feature    98..99
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     promoter        148..1194
                     /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"
     misc_feature    168..180
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     primer_bind     331..351
                     /%GC="57.1"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="4.4"
                     /Tm="60.0"
                     /annotation_group="60523053606300000814a7f4-bb8a-49a8-b7fa
                     -d26557e9de2a"
                     /created_by="primer3"
                     /Sequence="GATGGTTAGAGAGGCCTACGC"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-L4-2-R1"
                     /label="P10"
     primer_bind     817..836
                     /%GC="55.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="3.1"
                     /Tm="60.2"
                     /annotation_group="60523054200048009c5c0bec-d599-4051-8978
                     -7b3e8b59d21a"
                     /created_by="primer3"
                     /Sequence="CGGATTCCATTGCCCAGCTA"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-L4-2-R1"
                     /label="P11"
     misc_feature    929
                     /Original_Bases="T"
                     /label="T"
                     /note="Geneious type: Editing History Replacement"
     misc_feature    1124
                     /Original_Bases="G"
                     /label="G"
                     /note="Geneious type: Editing History Replacement"
     misc_feature    1126
                     /Original_Bases="A"
                     /label="A"
                     /note="Geneious type: Editing History Replacement"
     misc_feature    1127^1128
                     /Original_Bases="GACGATCCG"
                     /label="GACGATCCG"
                     /note="Geneious type: Editing History Deletion"
     misc_feature    1223
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     attR1           1224..1380
                     /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="attR1"
                     /Transferred_Similarity="99.36%"
                     /Primary_Match="99.36%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:y-f0o11e8"">attR1</a>"
     misc_feature    1350
                     /Original_Bases="T"
                     /label="T"
                     /note="Geneious type: Editing History Replacement"
     primer          complement(1443..1459)
                     /invitrogen=2010000
                     /note="Derived using Geneious Prime 2020.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="pDONRP4-P3 - M13 reverse primer"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; M13
                     reverse primer (pDONRP4-P3 - M13 reverse primer)"
                     /label="M13 reverse primer"
     promoter        1521..1571
                     /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    1572..2381
                     /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; <a
                     href=""urn:local:.:8j-dgycryy"">KanR</a>"
                     /label="KanR"
                     /note="Geneious type: Bacterial Selection"
     primer_bind     1597..1616
                     /%GC="55.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="25.9"
                     /Tm="59.8"
                     /annotation_group="60523057819143003871436a-6251-4c58-a35e
                     -437fb617fa5d"
                     /created_by="primer3"
                     /Sequence="CGAGGCCGCGATTAAATTCC"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-L4-2-R1"
                     /label="P44"
     primer_bind     2231..2250
                     /%GC="60.0"
                     /Hairpin_Tm="40.0"
                     /Self_Dimer_Tm="None"
                     /Tm="60.0"
                     /annotation_group="605230565714010024e5cde3-0cd1-4f10-b6fb
                     -3da691dda779"
                     /created_by="primer3"
                     /Sequence="CGCAGACCGATACCAGGATC"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-L4-2-R1"
                     /label="P42"
     rep_origin      2528..3116
                     /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="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    2675
                     /Original_Bases="C"
                     /label="C"
                     /note="Geneious type: Editing History Replacement"
     misc_feature    2920..2921
                     /Original_Bases="AT"
                     /label="AT"
                     /note="Geneious type: Editing History Replacement"
     primer_bind     3016..3035
                     /%GC="55.0"
                     /Hairpin_Tm="37.3"
                     /Self_Dimer_Tm="None"
                     /Tm="59.7"
                     /annotation_group="60523058493578006978c923-96f3-4d83-86b8
                     -72f3eca53f48"
                     /created_by="primer3"
                     /Sequence="GGGGAAACGCCTGGTATCTT"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-L4-2-R1"
                     /label="P45"
     primer_bind     3624..3643
                     /%GC="55.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="60.0"
                     /annotation_group="6052305907507000b5b5359e-3d25-4a5a-980f
                     -5bf90e9a5945"
                     /created_by="primer3"
                     /Sequence="CTTCCGACTGAGCCTTTCGT"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-L4-2-R1"
                     /label="P46"
     primer          3695..3711
                     /invitrogen=2000000
                     /note="Derived using Geneious Prime 2020.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="pDONRP4-P3 - M13 (-40) forward primer"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; M13
                     (-40) forward primer (pDONRP4-P3 - M13 (-40) forward
                     primer)"
                     /label="M13 (-40) forward primer"
     primer          3715..3730
                     /invitrogen=1980000
                     /note="Derived using Geneious Prime 2020.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="pDONRP4-P3 - M13 (-20) forward primer"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; M13
                     (-20) forward primer (pDONRP4-P3 - M13 (-20) forward
                     primer)"
                     /label="M13 (-20) forward primer"
     misc_feature    3748
                     /Original_Bases="C"
                     /label="C"
                     /note="Geneious type: Editing History Replacement"
ORIGIN      
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgataagcaa
       61 tgctttttta taatgccaac tttgtataga aaagttgtca tttaggtgac actatagaat
      121 actcaagcta tgcatccaac gcgttgggag ctctcccata tggtcgacta gagccaagct
      181 gatctccttt gccccggaga tcaccatgga cgactttctc tatctctacg atctaggaag
      241 aaagttcgac ggagaaggtg acgataccat gttcaccacc gataatgaga agattagcct
      301 cttcaatttc agaaagaatg ctgacccaca gatggttaga gaggcctacg cggcaggtct
      361 catcaagacg atctacccga gtaataatct ccaggagatc aaataccttc ccaagaaggt
      421 taaagatgca gtcaaaagat tcaggactaa ctgcatcaag aacacagaga aagatatatt
      481 tctcaagatc agaagtacta ttccagtatg gacgattcaa ggcttgcttc ataaaccaag
      541 gcaagtaata gagattggag tctctaagaa agtagttcct actgaatcaa aggccatgga
      601 gtcaaaaatt cagatcgagg atctaacaga actcgccgtg aagactggcg aacagttcat
      661 acagagtctt ttacgactca atgacaagaa gaaaatcttc gtcaacatgg tggagcacga
      721 cactctcgtc tactccaaga atatcaaaga tacagtctca gaagaccaaa gggctattga
      781 gacttttcaa caaagggtaa tatcgggaaa cctcctcgga ttccattgcc cagctatctg
      841 tcacttcatc aaaaggacag tagaaaagga aggtggcacc tacaaatgcc atcattgcga
      901 taaaggaaag gctatcgttc aagatgcccc tgccgacagt ggtcccaaag atggaccccc
      961 acccacgagg agcatcgtgg aaaaagaaga cgttccaacc acgtcttcaa agcaagtgga
     1021 ttgatgtgat atctccactg acgtaaggga tgacgcacaa tcccactatc cttcgcaaga
     1081 cccttcctct atataaggaa gttcatttca tttggagagg actccggtat ttttacaaca
     1141 attaccacaa caaaacaaac aacaaacaac attacaattt actattctag tcgacctgca
     1201 ggcggccgca ctagtgatat cagcaagttt gtacaaaaaa gttgaacgag aaacgtaaaa
     1261 tgatataaat atcaatatat taaattagat tttgcataaa aaacagacta cataatactg
     1321 taaaacacaa catatgcagt cactatgaac caactactta gatggtatta gtgacctgta
     1381 gaattcgagc tctagagctg cagggcggcc gcgatatccc ctatagtgag tcgtattaca
     1441 tggtcatagc tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg
     1501 cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata
     1561 caaggggtgt tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca
     1621 tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga
     1681 caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag
     1741 gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta
     1801 tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca
     1861 ctgcgatccc cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa
     1921 atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt
     1981 gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg
     2041 gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct
     2101 ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt
     2161 tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac
     2221 gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt
     2281 tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga
     2341 ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt
     2401 tgtaacactg gcagagcatt acgctgactt gacgggacgg cgcaagctca tgaccaaaat
     2461 cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     2521 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     2581 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     2641 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     2701 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     2761 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     2821 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     2881 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     2941 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     3001 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     3061 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     3121 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     3181 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     3241 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     3301 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     3361 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatacg cgtaccgcta
     3421 gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct tctgcttagt
     3481 ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc gttgcttcac
     3541 aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca ccgacaaaca
     3601 acagataaaa cgaaaggccc agtcttccga ctgagccttt cgttttattt gatgcctggc
     3661 agttccctac tctcgcgtta acgctagcat ggatgttttc ccagtcacga cgttgtaaaa
     3721 cgacggccag tcttaagctc gggcccgagt taacgctacc atggagctc
//
