LOCUS       pEN-L4-2-L3             3661 bp    DNA     circular     06-SEP-2022
DEFINITION  .
ACCESSION   urn.local...12f-ds878jq
KEYWORDS    .
SOURCE      
  ORGANISM  .
COMMENT     This file is created by Vector NTI                                 
            http://www.invitrogen.com/                                         
            ORIGDB|GenBank                                                     
            VNTDATE|461504715|                                                 
            VNTDBDATE|461504715|                                               
            LSOWNER|                                                           
            VNTNAME|pEN-L4-2-L3,0|                                             
            VNTAUTHORNAME|Jelle Van Leene|                                     
            VNTAUTHORTEL|0485280590|                                           
            VNTAUTHOREML|jelee@psb.ugent.be|                                   
            VNTAUTHORAD1|Technologiepark 927|                                  
            VNTAUTHORAD2|9000 Gent|                                            
            Vector_NTI_Display_Data_(Do_Not_Edit!)                             
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            0.123113 1)                          #233=(CLabel                  
                                                       (CWidget 0 (0 0) 1 2 0 0
            #170# 26089628 100)                                (LOGPEN 0 0
            8437824) 1                                              (LOGFONT 35
            13 0 0 400 255 0 0 0 3 2 1 34                                
            "Arial") 1.15807 0.611111 0                                       
            "{\\i Pst}I (1337)" "@N (@S)" 5 12.0352                            
               -11.5807 0 -10 2.34558 0.670166 #232#))                        
            (CObjectList))                                               
            #234=(CGroupWidget                                                 
                  (CWidget 2 (10 #16# 0) 1 2 0 0 Nil -1223 100)                
                    (CObjectList                                               
                     #235=(CScratch                                            
                           (CWidget 1 (1 #16# 1) 1 2 0 0 #128# 0 100)          
                                 (LOGPEN 0 8 0) 8 1 6.00221 1.9 0.123113 1)    
                                 #236=(CLabel                                  
                                       (CWidget 0 (0 0) 1 2 0 0 #170# 26660924
            100)                                (LOGPEN 0 0 8437824) 1         
                                                (LOGFONT 35 13 0 0 400 255 0 0
            0 3 2 1 34                                 "Arial") 1.15807
            0.611111 0                                        "{\\i Sal}I
            (161)" "@N (@S)" 5 7.80408                                11.5807 0
            -10 2.10905 0.670166 #235#)                          #237=(CScratch
                                                                       (CWidget
            2 (1 #16# 2) 1 2 0 0 #128# 0 100)                               
            (LOGPEN 0 8 0) 8 1 4.26455 1.9 0.123113 1)                         
            #238=(CLabel                                                       
                  (CWidget 0 (0 0) 1 2 0 0 #170# 21427860 100)                 
                          (LOGPEN 0 0 8437824) 1                               
                          (LOGFONT 35 13 0 0 400 255 0 0 0 3 2 1 34            
                                "Arial") 1.15807 0.611111 0                    
                               "{\\i Sal}I (1187)" "@N (@S)" 5 14.0631         
                                  -3.47421 0 -10 2.34558 0.670166 #237#))      
                              (CObjectList))) (CObjectList))                   
                 #239=(CGroupWidget (CWidget 14 (16 0) 1 2 0 0 Nil -519 100)   
                          (CObjectList) (CObjectList))                         
                    #240=(CGroupWidget (CWidget 12 (0 0) 1 2 0 0 Nil -637 100) 
                            (CObjectList) (CObjectList))) (CObjectList)))      
              (CSeqView 10 5 (CObjectList) (CObList) 1 (CObList)) (CObList)
            1953068654    (CStringList) 1952671094 891318895 (CObList))).
FEATURES             Location/Qualifiers
     misc_feature    0^1
                     /Original_Bases="GCGTTAACGCTACCATGGAGCTC"
                     /label="GCGTTAACGCTACCATGGAGCTC"
                     /note="Geneious type: Editing History Deletion"
     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="98.97%"
                     /Primary_Match="98.97%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:lr-eztz0t4"">attL4</a>"
     misc_feature    55
                     /Original_Bases="A"
                     /label="A"
                     /note="Geneious type: Editing History Replacement"
     promoter        146..1192
                     /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    183
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     primer_bind     329..349
                     /%GC="57.1"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="4.4"
                     /Tm="60.0"
                     /annotation_group="605697738235540039477a8d-1cda-4339-ba01
                     -7daafd89fc06"
                     /created_by="primer3"
                     /Sequence="GATGGTTAGAGAGGCCTACGC"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-L4-2-L3"
                     /label="P10"
     misc_feature    927
                     /Original_Bases="T"
                     /label="T"
                     /note="Geneious type: Editing History Replacement"
     primer_bind     1071..1090
                     /%GC="50.0"
                     /Hairpin_Tm="45.0"
                     /Self_Dimer_Tm="None"
                     /Tm="57.8"
                     /annotation_group="6056977973511400b7260e62-db14-4fd3-8280
                     -1193fe1064c7"
                     /created_by="primer3"
                     /Sequence="TCGCAAGACCCTTCCTCTAT"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-L4-2-L3"
                     /label="P64"
     attL3           complement(1221..1316)
                     /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>"
     misc_feature    1318..1327
                     /Original_Bases="AATTCGAGCTCTAGAGCTGCAGGGCGGCCGC"
                     /label="AATTCGAGCTCTAGAGCTGCAGGGCGGCCGC"
                     /note="Geneious type: Editing History Replacement"
     primer          complement(1355..1375)
                     /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        1436..1486
                     /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    1487..2296
                     /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     1512..1531
                     /%GC="55.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="25.9"
                     /Tm="59.8"
                     /annotation_group="60569778000828006090c24d-eb71-4c99-92b3
                     -da2e725490a6"
                     /created_by="primer3"
                     /Sequence="CGAGGCCGCGATTAAATTCC"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-L4-2-L3"
                     /label="P44"
     primer_bind     2146..2165
                     /%GC="60.0"
                     /Hairpin_Tm="40.0"
                     /Self_Dimer_Tm="None"
                     /Tm="60.0"
                     /annotation_group="605697768422310075908342-0c58-4104-9923
                     -110fc434e29e"
                     /created_by="primer3"
                     /Sequence="CGCAGACCGATACCAGGATC"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-L4-2-L3"
                     /label="P42"
     rep_origin      2443..3031
                     /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    2590
                     /Original_Bases="C"
                     /label="C"
                     /note="Geneious type: Editing History Replacement"
     misc_feature    2835..2836
                     /Original_Bases="AT"
                     /label="AT"
                     /note="Geneious type: Editing History Replacement"
     primer_bind     2931..2950
                     /%GC="55.0"
                     /Hairpin_Tm="37.3"
                     /Self_Dimer_Tm="None"
                     /Tm="59.7"
                     /annotation_group="60569778571346004f378156-7943-459a-989e
                     -3e673aaf6794"
                     /created_by="primer3"
                     /Sequence="GGGGAAACGCCTGGTATCTT"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-L4-2-L3"
                     /label="P45"
     primer_bind     3539..3558
                     /%GC="55.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="60.0"
                     /annotation_group="605697791416220055670301-ce7d-4461-82cb
                     -17a9386a469e"
                     /created_by="primer3"
                     /Sequence="CTTCCGACTGAGCCTTTCGT"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-L4-2-L3"
                     /label="P46"
     primer          3630..3650
                     /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"
ORIGIN      
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgatgagcaa
       61 tgctttttta taatgccaac tttgtataga aaagttgatt taggtgacac tatagaatac
      121 tcaagctatg catccaacgc gttgggagct ctcccatatg gtcgactaga gccaagctga
      181 tctcctttgc cccggagatc accatggacg actttctcta tctctacgat ctaggaagaa
      241 agttcgacgg agaaggtgac gataccatgt tcaccaccga taatgagaag attagcctct
      301 tcaatttcag aaagaatgct gacccacaga tggttagaga ggcctacgcg gcaggtctca
      361 tcaagacgat ctacccgagt aataatctcc aggagatcaa ataccttccc aagaaggtta
      421 aagatgcagt caaaagattc aggactaact gcatcaagaa cacagagaaa gatatatttc
      481 tcaagatcag aagtactatt ccagtatgga cgattcaagg cttgcttcat aaaccaaggc
      541 aagtaataga gattggagtc tctaagaaag tagttcctac tgaatcaaag gccatggagt
      601 caaaaattca gatcgaggat ctaacagaac tcgccgtgaa gactggcgaa cagttcatac
      661 agagtctttt acgactcaat gacaagaaga aaatcttcgt caacatggtg gagcacgaca
      721 ctctcgtcta ctccaagaat atcaaagata cagtctcaga agaccaaagg gctattgaga
      781 cttttcaaca aagggtaata tcgggaaacc tcctcggatt ccattgccca gctatctgtc
      841 acttcatcaa aaggacagta gaaaaggaag gtggcaccta caaatgccat cattgcgata
      901 aaggaaaggc tatcgttcaa gatgcccctg ccgacagtgg tcccaaagat ggacccccac
      961 ccacgaggag catcgtggaa aaagaagacg ttccaaccac gtcttcaaag caagtggatt
     1021 gatgtgatat ctccactgac gtaagggatg acgcacaatc ccactatcct tcgcaagacc
     1081 cttcctctat ataaggaagt tcatttcatt tggagaggac tccggtattt ttacaacaat
     1141 taccacaaca aaacaaacaa caaacaacat tacaatttac tattctagtc gacctgcagg
     1201 cggccgcact agtgatatca caactttatt atacaaagtt ggcattataa aaaagcattg
     1261 cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca
     1321 tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct
     1381 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
     1441 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
     1501 acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa
     1561 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc
     1621 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
     1681 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
     1741 tatccgtact cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt
     1801 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
     1861 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
     1921 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
     1981 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc
     2041 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     2101 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     2161 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     2221 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     2281 cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct
     2341 gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt
     2401 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     2461 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     2521 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     2581 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     2641 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     2701 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     2761 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     2821 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     2881 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     2941 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     3001 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     3061 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     3121 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     3181 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     3241 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     3301 cagtgagcgc aacgcaatta atacgcgtac cgctagccag gaagagtttg tagaaacgca
     3361 aaaaggccat ccgtcaggat ggccttctgc ttagtttgat gcctggcagt ttatggcggg
     3421 cgtcctgccc gccaccctcc gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga
     3481 tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct
     3541 tccgactgag cctttcgttt tatttgatgc ctggcagttc cctactctcg cgttaacgct
     3601 agcatggatg ttttcccagt cacgacgttg taaaacgacg gccagtctta agctcgggcc
     3661 c
//
