LOCUS       pEN-R2-S*-L3            4454 bp    DNA     circular     12-JUL-2022
DEFINITION  .
ACCESSION   urn.local...7e-dv3ipbw
KEYWORDS    .
SOURCE      
  ORGANISM  .
COMMENT     This file is created by Vector NTI                                 
            http://www.invitrogen.com/                                         
            VNTDATE|459289516|                                                 
            VNTDBDATE|459289576|                                               
            LSOWNER|                                                           
            VNTNAME|pEN-R2-S*-L3,0|                                            
            VNTAUTHORNAME|Demo User|                                           
            VNTAUTHOREML|manka@psb.ugent.be|                                   
            VNTOAUTHORNAME|Mansour Karimi|                                     
            VNTOAUTHORTEL|+3193313850|                                         
            VNTOAUTHOREML|mansour.karimi@psb.ugent.be|                         
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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..1939
                     /modified_by="mamee"
                     /label="GUS*"
     primer_bind     217..236
                     /%GC="50.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="59.9"
                     /annotation_group="60530271695580000e87fb3d-d751-4aad-83df
                     -9cd0653adfe0"
                     /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="605302722845520031b94c6c-2b52-4e8a-9292
                     -4e5bfeeeae15"
                     /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="60530272861610007a02022d-f1f8-415a-8d3e
                     -a12a441bf188"
                     /created_by="primer3"
                     /Sequence="GGAAGCAACGCGTAAACTCG"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-R2-S*-L3"
                     /label="P32"
     attL3           complement(1941..2036)
                     /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     2061..2080
                     /%GC="60.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="8.2"
                     /Tm="59.3"
                     /annotation_group="6053027405166100d56a579c-1c2d-44ed-93dc
                     -7c2931a9d8fe"
                     /created_by="primer3"
                     /Sequence="GGCCGCGATATCCCCTATAG"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-R2-S*-L3"
                     /label="P40"
     primer          complement(2097..2113)
                     /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        2175..2225
                     /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    2226..3035
                     /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     2885..2904
                     /%GC="60.0"
                     /Hairpin_Tm="40.0"
                     /Self_Dimer_Tm="None"
                     /Tm="60.0"
                     /annotation_group="605302746209350001e9f8a6-d000-4f80-9753
                     -6eced3b4f6bb"
                     /created_by="primer3"
                     /Sequence="CGCAGACCGATACCAGGATC"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-R2-S*-L3"
                     /label="P42"
     rep_origin      3182..3770
                     /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    3329
                     /Original_Bases="C"
                     /label="C"
                     /note="Geneious type: Editing History Replacement"
     misc_feature    3574..3575
                     /Original_Bases="AT"
                     /label="AT"
                     /note="Geneious type: Editing History Replacement"
     primer_bind     3670..3689
                     /%GC="55.0"
                     /Hairpin_Tm="37.3"
                     /Self_Dimer_Tm="None"
                     /Tm="59.7"
                     /annotation_group="605302763475120058a9bc9f-a0f2-432d-9f13
                     -2182088b29d2"
                     /created_by="primer3"
                     /Sequence="GGGGAAACGCCTGGTATCTT"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-R2-S*-L3"
                     /label="P45"
     primer_bind     4278..4297
                     /%GC="55.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="60.0"
                     /annotation_group="60530276937396008b4f8bfa-152b-4fcd-af24
                     -5d77778179b9"
                     /created_by="primer3"
                     /Sequence="CTTCCGACTGAGCCTTTCGT"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-R2-S*-L3"
                     /label="P46"
     primer          4349..4365
                     /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          4369..4384
                     /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 aaacaatgaa caactttatt atacaaagtt ggcattataa aaaagcattg
     1981 cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttggag
     2041 ctccatggta gcgttaacgc ggccgcgata tcccctatag tgagtcgtat tacatggtca
     2101 tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag
     2161 ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg
     2221 gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg
     2281 ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct
     2341 atcgcttgta tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg
     2401 ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc
     2461 ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga
     2521 tccccggaaa aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg
     2581 ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt
     2641 ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg
     2701 ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag
     2761 aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac
     2821 ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg
     2881 gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc
     2941 cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat
     3001 tgcagtttca tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac
     3061 actggcagag cattacgctg acttgacggg acggcgcaag ctcatgacca aaatccctta
     3121 acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     3181 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     3241 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     3301 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     3361 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     3421 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     3481 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     3541 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     3601 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     3661 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     3721 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     3781 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     3841 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     3901 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     3961 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     4021 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc gctagccagg
     4081 aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg gccttctgct tagtttgatg
     4141 cctggcagtt tatggcgggc gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt
     4201 caaatccgct cccggcggat ttgtcctact caggagagcg ttcaccgaca aacaacagat
     4261 aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt atttgatgcc tggcagttcc
     4321 ctactctcgc gttaacgcta gcatggatgt tttcccagtc acgacgttgt aaaacgacgg
     4381 ccagtcttaa gctcgggccc tgcagctcta gagctcgaat tctacaggtc actaatacca
     4441 tctaagtagt tggt
//
