LOCUS       pEN-R2-S-L3             4451 bp    DNA     circular     12-JUL-2022
DEFINITION  .
ACCESSION   urn.local...7t-dv3j7dn
KEYWORDS    .
SOURCE      
  ORGANISM  .
COMMENT     This file is created by Vector NTI                                 
            http://www.invitrogen.com/                                         
            VNTDATE|458702480|                                                 
            VNTDBDATE|459206801|                                               
            LSOWNER|                                                           
            VNTNAME|pEN-R2SL3,0|                                               
            VNTAUTHORNAME|Demo User|                                           
            VNTAUTHOREML|manka@psb.ugent.be|                                   
            VNTOAUTHORNAME|Mansour Karimi|                                     
            VNTOAUTHORTEL|+3193313850|                                         
            VNTOAUTHOREML|mansour.karimi@psb.ugent.be|                         
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                     -13.8968 0 -10 2.73968 0.676024 #227#))                   
                 (CObjectList))                                               
            #229=(CGroupWidget                                                 
                  (CWidget 2 (10 #14# 0) 1 2 0 0 Nil -745 100)                 
                   (CObjectList                                                
                    #230=(CScratch                                             
                          (CWidget 1 (1 #14# 1) 1 2 0 0 #163# 0 100)           
                                (LOGPEN 0 0 10079283) 1 1 2.95408 1.9          
                                 0.123113 1)                                   
                           #231=(CLabel                                        
                                 (CWidget 0 (0 0) 1 2 0 0 #151# 14352800 100)  
                                         (LOGPEN 0 0 8437824) 1                
                                         (LOGFONT 39 14 0 0 400 255 0 0 0 3 2 1
            34                                 "Arial") 1.15807 0.611111 0     
                                              "{\\i Sac}I (2356)" "@N (@S)" 5
            -4.80317                                -12.7388 0 -10 2.73968
            0.676024 #230#)                          #232=(CScratch            
                                                           (CWidget 2 (1 #14#
            2) 1 2 0 0 #163#                                  1196708929 100)
            (LOGPEN 0 0 10079283) 1 1                                0.025369
            1.9 0.123113 1)                                      #233=(CLabel  
                                                                       (CWidget
            0 (0 0) 1 2 0 0 #151# 2082602612 100)                              
             (LOGPEN 0 0 8437824) 1                                            
             (LOGFONT 39 14 0 0 400 255 0 0 0 3 2 1 34                         
                   "Arial") 1.15807 0.611111 0                                 
                  "{\\i Sac}I (4434)" "@N (@S)" 5 -4.74358                     
                      12.7388 0 -10 2.73968 0.676024 #232#))                   
                 (CObjectList))) (CObjectList))                        
            #234=(CGroupWidget (CWidget 14 (16 0) 1 2 0 0 Nil -781 100)        
                     (CObjectList) (CObjectList))                              
               #235=(CGroupWidget (CWidget 12 (0 0) 1 2 0 0 Nil 1635017063 100)
                             (CObjectList) (CObjectList))) (CObjectList)))     
               (CSeqView 10 5 (CObjectList) (CObList) 1 (CObList)) (CObList)
            27550216    (CStringList) 34078733 14353176 (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>"
     CDS             128..1936
                     /label="GUS"
     primer_bind     217..236
                     /%GC="50.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="59.9"
                     /annotation_group="6053060356573000ed4bb1ce-1742-44c5-af9d
                     -cf0685c5ab86"
                     /created_by="primer3"
                     /Sequence="TGATCAGCGTTGGTGGGAAA"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-R2-S-L3"
                     /label="P30"
     primer_bind     826..845
                     /%GC="50.0"
                     /Hairpin_Tm="46.3"
                     /Self_Dimer_Tm="None"
                     /Tm="59.7"
                     /annotation_group="605306041532450080913e23-9f10-4abd-9920
                     -7f9d47ec3734"
                     /created_by="primer3"
                     /Sequence="TTTGCAAGTGGTGAATCCGC"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-R2-S-L3"
                     /label="P31"
     primer_bind     1414..1433
                     /%GC="55.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="14.5"
                     /Tm="60.2"
                     /annotation_group="605306047336890000b1ff59-b2d4-4c84-8124
                     -352a89e52211"
                     /created_by="primer3"
                     /Sequence="GGAAGCAACGCGTAAACTCG"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-R2-S-L3"
                     /label="P32"
     attL3           complement(1938..2033)
                     /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_bind     2058..2077
                     /%GC="60.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="8.2"
                     /Tm="59.3"
                     /annotation_group="6053060591737300d957a957-6947-43f7-a066
                     -eaa015a3d9ae"
                     /created_by="primer3"
                     /Sequence="GGCCGCGATATCCCCTATAG"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-R2-S-L3"
                     /label="P40"
     primer          complement(2094..2110)
                     /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        2172..2222
                     /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    2223..3032
                     /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     2882..2901
                     /%GC="60.0"
                     /Hairpin_Tm="40.0"
                     /Self_Dimer_Tm="None"
                     /Tm="60.0"
                     /annotation_group="6053060651086800f9324046-7d5d-4c29-b57d
                     -7025045255ab"
                     /created_by="primer3"
                     /Sequence="CGCAGACCGATACCAGGATC"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-R2-S-L3"
                     /label="P42"
     rep_origin      3179..3767
                     /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    3326
                     /Original_Bases="C"
                     /label="C"
                     /note="Geneious type: Editing History Replacement"
     misc_feature    3571..3572
                     /Original_Bases="AT"
                     /label="AT"
                     /note="Geneious type: Editing History Replacement"
     primer_bind     3667..3686
                     /%GC="55.0"
                     /Hairpin_Tm="37.3"
                     /Self_Dimer_Tm="None"
                     /Tm="59.7"
                     /annotation_group="605306082444250026b51d17-039a-475d-9aea
                     -7cc45b8e997a"
                     /created_by="primer3"
                     /Sequence="GGGGAAACGCCTGGTATCTT"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-R2-S-L3"
                     /label="P45"
     primer_bind     4275..4294
                     /%GC="55.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="60.0"
                     /annotation_group="60530608819758002ce0e353-592d-444f-8a9d
                     -1836c948979a"
                     /created_by="primer3"
                     /Sequence="CTTCCGACTGAGCCTTTCGT"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-R2-S-L3"
                     /label="P46"
     primer          4346..4362
                     /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          4366..4381
                     /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"
ORIGIN      
        1 tcatagtgac tgcatatgtt gtgttttaca gtattatgta gtctgttttt tatgcaaaat
       61 ctaatttaat atattgatat ttatatcatt ttacgtttct cgttcaactt tcttgtacaa
      121 agtgggaatg ttacgtcctg tagaaacccc aacccgtgaa atcaaaaaac tcgacggcct
      181 gtgggcattc agtctggatc gcgaaaactg tggaattgat cagcgttggt gggaaagcgc
      241 gttacaagaa agccgggcaa ttgctgtgcc aggcagtttt aacgatcagt tcgccgatgc
      301 agatattcgt aattatgcgg gcaacgtctg gtatcagcgc gaagtcttta taccgaaagg
      361 ttgggcaggc cagcgtatcg tgctgcgttt cgatgcggtc actcattacg gcaaagtgtg
      421 ggtcaataat caggaagtga tggagcatca gggcggctat acgccatttg aagccgatgt
      481 cacgccgtat gttattgccg ggaaaagtgt acgtatcacc gtttgtgtga acaacgaact
      541 gaactggcag actatcccgc cgggaatggt gattaccgac gaaaacggca agaaaaagca
      601 gtcttacttc catgatttct ttaactatgc cggaatccat cgcagcgtaa tgctctacac
      661 cacgccgaac acctgggtgg acgatatcac cgtggtgacg catgtcgcgc aagactgtaa
      721 ccacgcgtct gttgactggc aggtggtggc caatggtgat gtcagcgttg aactgcgtga
      781 tgcggatcaa caggtggttg caactggaca aggcactagc gggactttgc aagtggtgaa
      841 tccgcacctc tggcaaccgg gtgaaggtta tctctatgaa ctgtgcgtca cagccaaaag
      901 ccagacagag tgtgatatct acccgcttcg cgtcggcatc cggtcagtgg cagtgaaggg
      961 cgaacagttc ctgattaacc acaaaccgtt ctactttact ggctttggtc gtcatgaaga
     1021 tgcggacttg cgtggcaaag gattcgataa cgtgctgatg gtgcacgacc acgcattaat
     1081 ggactggatt ggggccaact cctaccgtac ctcgcattac ccttacgctg aagagatgct
     1141 cgactgggca gatgaacatg gcatcgtggt gattgatgaa actgctgctg tcggctttaa
     1201 cctctcttta ggcattggtt tcgaagcggg caacaagccg aaagaactgt acagcgaaga
     1261 ggcagtcaac ggggaaactc agcaagcgca cttacaggcg attaaagagc tgatagcgcg
     1321 tgacaaaaac cacccaagcg tggtgatgtg gagtattgcc aacgaaccgg atacccgtcc
     1381 gcaaggtgca cgggaatatt tcgcgccact ggcggaagca acgcgtaaac tcgacccgac
     1441 gcgtccgatc acctgcgtca atgtaatgtt ctgcgacgct cacaccgata ccatcagcga
     1501 tctctttgat gtgctgtgcc tgaaccgtta ttacggatgg tatgtccaaa gcggcgattt
     1561 ggaaacggca gagaaggtac tggaaaaaga acttctggcc tggcaggaga aactgcatca
     1621 gccgattatc atcaccgaat acggcgtgga tacgttagcc gggctgcact caatgtacac
     1681 cgacatgtgg agtgaagagt atcagtgtgc atggctggat atgtatcacc gcgtctttga
     1741 tcgcgtcagc gccgtcgtcg gtgaacaggt atggaatttc gccgattttg cgacctcgca
     1801 aggcatattg cgcgttggcg gtaacaagaa agggatcttc actcgcgacc gcaaaccgaa
     1861 gtcggcggct tttctgctgc aaaaacgctg gactggcatg aacttcggtg aaaaaccgca
     1921 gcagggaggc aaacaaacaa ctttattata caaagttggc attataaaaa agcattgctt
     1981 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttggagctc
     2041 catggtagcg ttaacgcggc cgcgatatcc cctatagtga gtcgtattac atggtcatag
     2101 ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata
     2161 aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg
     2221 ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg
     2281 atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc
     2341 gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg
     2401 ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc
     2461 cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc
     2521 ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg
     2581 atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta
     2641 acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg
     2701 atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa
     2761 tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg
     2821 ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa
     2881 tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt
     2941 cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc
     3001 agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact
     3061 ggcagagcat tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg
     3121 tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     3181 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     3241 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     3301 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     3361 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     3421 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     3481 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     3541 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     3601 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     3661 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     3721 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     3781 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     3841 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     3901 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     3961 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     4021 ccgactggaa agcgggcagt gagcgcaacg caattaatac gcgtaccgct agccaggaag
     4081 agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttag tttgatgcct
     4141 ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca caacgttcaa
     4201 atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa
     4261 acgaaaggcc cagtcttccg actgagcctt tcgttttatt tgatgcctgg cagttcccta
     4321 ctctcgcgtt aacgctagca tggatgtttt cccagtcacg acgttgtaaa acgacggcca
     4381 gtcttaagct cgggccctgc agctctagag ctcgaattct acaggtcact aataccatct
     4441 aagtagttgg t
//
