LOCUS       pEN-R2-3xFlag-L3        2723 bp    DNA     circular     06-SEP-2022
DEFINITION  .
ACCESSION   urn.local...5h-dv8t2tn
KEYWORDS    .
SOURCE      
  ORGANISM  .
COMMENT     http://www.informaxinc.com/                                        
            This file is created by Vector NTI                                 
            http://www.invitrogen.com/                                         
            ORIGDB|GenBank                                                     
            VNTDATE|470845167|                                                 
            VNTDBDATE|470845167|                                               
            LSOWNER|                                                           
            VNTNAME|pEN-R2-3xFlag-L3|                                          
            VNTAUTHORNAME|Mansour Karimi|                                      
            VNTAUTHORTEL|003293313838|                                         
            VNTAUTHOREML|mansour.karimi@psb.ugent.be|                          
            VNTOAUTHORNAME|Computer Group|                                     
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                                                (LOGFONT 35 13 0 0 400 255 1 0
            0 3 2 1 34                                 "Arial") 1.15807
            0.611111 0                                        "{\\i Nru}I
            (362)" "@N (@S)" 5 11.9355                                8.10649 0
            -10 2.24703 0.670166 #235#))                         (CObjectList))
                                                           #237=(CGroupWidget  
                                                                 (CWidget 2 (10
            #9# 0) 1 2 0 0 Nil -989 100)                         (CObjectList  
                                                                 
            #238=(CScratch                                                     
                  (CWidget 1 (1 #9# 1) 1 2 0 0 #131# 0 100)                    
                       (LOGPEN 0 8 0) 8 1 6.05441 1.9 0.123113 1)              
                       #239=(CLabel (CWidget 0 (0 0) 1 2 0 0 #184# 0 100)      
                                     (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 Sac}I (96)" "@N (@S)" 5 5.07849
            13.8968                                0 -10 1.99079 0.670166
            #238#)                                 #240=(CScratch              
                                                         (CWidget 2 (1 #9# 2) 1
            2 0 0 #131# 0 100)                                (LOGPEN 0 8 0) 8
            1 0.58955 1.9 0.123113 1)                          #241=(CLabel    
                                                                     (CWidget 0
            (0 0) 1 2 0 0 #184# 28334116 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 Sac}I (2469)" "@N (@S)" 5 -9.50744                      
                     11.5807 0 -10 2.48356 0.670166 #240#))                    
                (CObjectList))) (CObjectList))                        
            #242=(CGroupWidget (CWidget 14 (16 0) 1 2 0 0 Nil -964 100)        
                     (CObjectList) (CObjectList))                              
               #243=(CGroupWidget (CWidget 12 (0 0) 1 2 0 0 Nil -108 100)      
                       (CObjectList) (CObjectList))) (CObjectList)))        
            (CSeqView 10 5 (CObjectList) (CObList) 1 (CObList)) (CObList) 0    
            (CStringList) 0 0 (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>"
     Linker          126..139
                     /modified_by="mamee"
                     /label="Gly4Ser-linker"
     misc_feature    126^127
                     /Original_Bases="G"
                     /label="G"
                     /note="Geneious type: Editing History Deletion"
     CDS             140..208
                     /label="3xFlag"
     attL3           complement(210..305)
                     /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(366..382)
                     /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        444..494
                     /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    495..1304
                     /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     520..539
                     /%GC="55.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="25.9"
                     /Tm="59.8"
                     /annotation_group="60572728146475002dc56903-d868-43c5-9182
                     -a1b4434c6f7e"
                     /created_by="primer3"
                     /Sequence="CGAGGCCGCGATTAAATTCC"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-R2-3xFlag-L3"
                     /label="P44"
     primer_bind     1154..1173
                     /%GC="60.0"
                     /Hairpin_Tm="40.0"
                     /Self_Dimer_Tm="None"
                     /Tm="60.0"
                     /annotation_group="6057272698591700ca5311b9-a90b-461a-a575
                     -80204697c88f"
                     /created_by="primer3"
                     /Sequence="CGCAGACCGATACCAGGATC"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-R2-3xFlag-L3"
                     /label="P42"
     rep_origin      1451..2039
                     /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="99.49%"
                     /Primary_Match="99.49%; 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    1598
                     /Original_Bases="C"
                     /label="C"
                     /note="Geneious type: Editing History Replacement"
     misc_feature    1843..1844
                     /Original_Bases="AT"
                     /label="AT"
                     /note="Geneious type: Editing History Replacement"
     primer_bind     1939..1958
                     /%GC="55.0"
                     /Hairpin_Tm="37.3"
                     /Self_Dimer_Tm="None"
                     /Tm="59.7"
                     /annotation_group="6057272873293000a6beeaf2-7906-4e5a-b85a
                     -bf176fe4d71a"
                     /created_by="primer3"
                     /Sequence="GGGGAAACGCCTGGTATCTT"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-R2-3xFlag-L3"
                     /label="P45"
     primer_bind     2547..2566
                     /%GC="55.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="60.0"
                     /annotation_group="605727293528760097512d35-a255-4782-b158
                     -e6361f5476b2"
                     /created_by="primer3"
                     /Sequence="CTTCCGACTGAGCCTTTCGT"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-R2-3xFlag-L3"
                     /label="P46"
     primer          2618..2634
                     /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          2638..2653
                     /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    2722
                     /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 agtgggtgga ggcggttcag actacaaaga ccatgacggt gattataaag atcatgacat
      181 cgactacaag gatgacgatg acaagtgatc aactttatta tacaaagttg gcattataaa
      241 aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat
      301 tatttggagc tccatggtag cgttaacgcg gccgcgatat cccctatagt gagtcgtatt
      361 acatggtcat agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca
      421 ttgcacaaga taaaaatata tcatcatgaa caataaaact gtctgcttac ataaacagta
      481 atacaagggg tgttatgagc catattcaac gggaaacgtc gaggccgcga ttaaattcca
      541 acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg
      601 cgacaatcta tcgcttgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca
      661 aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat
      721 ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca
      781 ccactgcgat ccccggaaaa acagcattcc aggtattaga agaatatcct gattcaggtg
      841 aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta
      901 attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata
      961 acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag
     1021 tctggaaaga aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg
     1081 atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg
     1141 gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg
     1201 agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata
     1261 tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt
     1321 ggttgtaaca ctggcagagc attacgctga cttgacggga cggcgcaagc tcatgaccaa
     1381 aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca
     1441 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     1501 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     1561 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     1621 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     1681 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     1741 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     1801 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     1861 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     1921 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     1981 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     2041 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     2101 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     2161 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     2221 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     2281 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg
     2341 ctagccagga agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt
     2401 agtttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt
     2461 cacaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa
     2521 acaacagata aaacgaaagg cccagtcttc cgactgagcc tttcgtttta tttgatgcct
     2581 ggcagttccc tactctcgcg ttaacgctag catggatgtt ttcccagtca cgacgttgta
     2641 aaacgacggc cagtcttaag ctcgggccct gcagctctag agctcgaatt ctacaggtca
     2701 ctaataccat ctaagtagtt ggt
//
