LOCUS       pEN-L1-LHGROATG4-L2        5599 bp    DNA     circular     06-SEP-2022
DEFINITION  .
ACCESSION   urn.local...ce-dstvtwn
KEYWORDS    .
SOURCE      
  ORGANISM  .
COMMENT     This file is created by Vector NTI                                 
            http://www.invitrogen.com/                                         
            VNTDATE|521742778|                                                 
            VNTDBDATE|521742778|                                               
            LSOWNER|                                                           
            VNTNAME|pEN-L1- LHGROATG4-L2|                                      
            VNTAUTHORNAME|Mansour Karimi|                                      
            VNTAUTHORTEL|003293313838|                                         
            VNTAUTHOREML|mansour.karimi@psb.ugent.be|                          
            VNTAUTHORWWW|http://www.psb.ugent.be|                              
            Vector_NTI_Display_Data_(Do_Not_Edit!)                             
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            #7=(CRSite (CStringList) "EcoRV" "gatatc" 4 0 2 897 0 5042 0 0 "") 
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            "SacII" "ccgcgg" 5 0 0 0 "")              #16=(CRSite (CStringList)
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                                                 #32=(CFSignal (CObList) "rrnB
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FEATURES             Location/Qualifiers
     attL1           1..100
                     /Color="#00cccc"
                     /created_by="mamee"
                     /note="Derived using Geneious Prime 2022.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="attL1"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:li-eztyqs1"">attL1</a>"
     misc_feature    80
                     /Original_Bases="G"
                     /label="G"
                     /note="Geneious type: Editing History Replacement"
     misc_feature    101
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     CDS             112..978
                     /label="GR"
     primer_bind     675..695
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-L1-LHGROATG4-L2"
                     /Sequence="GGAAGGTCTGAAGAGCCAAGA"
                     /Tm="59.4"
                     /%GC="52.4"
                     /Hairpin_Tm="43.1"
                     /Self_Dimer_Tm="5.9"
                     /modified_by="mamee"
                     /label="P61"
     CDS             1000..1965
                     /label="LacI"
     primer_bind     1461..1481
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-L1-LHGROATG4-L2"
                     /Sequence="TTTCTCCCATGAAGACGGTAC"
                     /Tm="57.4"
                     /%GC="47.6"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /modified_by="mamee"
                     /label="P62"
     misc_feature    1838
                     /Original_Bases="C"
                     /label="C"
                     /note="Geneious type: Editing History Replacement"
     CDS             1966..2326
                     /label="ATOG4"
     primer_bind     2071..2090
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-L1-LHGROATG4-L2"
                     /Sequence="ACTCAGACCAACTCTCAGGC"
                     /Tm="59.0"
                     /%GC="55.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /modified_by="mamee"
                     /label="P63"
     misc_feature    2327..2329
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     attL2           complement(2330..2429)
                     /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="attL2"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:ll-eztyzh8"">attL2</a>"
     misc_feature    2352
                     /Original_Bases="G"
                     /label="G"
                     /note="Geneious type: Editing History Replacement"
     promoter        2501..2551
                     /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"
     primer_bind     2577..2596
                     /%GC="55.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="25.9"
                     /Tm="59.8"
                     /annotation_group="60539447350608005ce4a136-96f4-42ef-b3d1
                     -6554641ee34d"
                     /created_by="primer3"
                     /Sequence="CGAGGCCGCGATTAAATTCC"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-L1-LHGROATG4-L2"
                     /label="P44"
     misc_feature    complement(3212..3745)
                     /note="Derived using Geneious Prime 2020.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /X-Transferred_From="GmR"
                     /X-Transferred_Similarity="100.00%"
                     /X-Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:8k-dgycryy"">GmR</a>"
                     /modified_by="mamee"
                     /note="Derived using Geneious Prime 2022.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="GmR"
                     /Transferred_Similarity="99.81%"
                     /Primary_Match="99.81%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:2d-f0u4295"">GmR</a>"
                     /label="GmR"
                     /note="Geneious type: Bacterial Selection"
     primer_bind     3274..3293
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-L1-LHGROATG4-L2"
                     /Sequence="TATAGAGAGCCACTGCGGGA"
                     /Tm="59.8"
                     /%GC="55.0"
                     /Hairpin_Tm="44.3"
                     /Self_Dimer_Tm="None"
                     /modified_by="mamee"
                     /label="P59"
     misc_feature    3453
                     /Original_Bases="C"
                     /label="C"
                     /note="Geneious type: Editing History Replacement"
     promoter        complement(3934..3961)
                     /invitrogen=2250000
                     /vntifkey=29
                     /note="Derived using Geneious Prime 2020.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /X-Transferred_From="pDEST32-GG"
                     /X-Transferred_Similarity="100.00%"
                     /X-Primary_Match="100.00%; Adjusted-Interval-0:0/0; Pc
                     promoter (<a
                     href=""urn:local:mamee:405-esp0pt5"">pDEST32-GG</a>)"
                     /modified_by="mamee"
                     /note="Derived using Geneious Prime 2022.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="GmR"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; pc (<a
                     href=""urn:local:.:2d-f0u4295"">GmR</a>)"
                     /label="pc"
     primer_bind     4077..4095
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-L1-LHGROATG4-L2"
                     /Sequence="CACGCATCGTCAGGCATTG"
                     /Tm="59.9"
                     /%GC="57.9"
                     /Hairpin_Tm="38.3"
                     /Self_Dimer_Tm="None"
                     /modified_by="mamee"
                     /label="P60"
     rep_origin      4619..5207
                     /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"
     primer_bind     4689..4708
                     /%GC="50.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="5.1"
                     /Tm="60.0"
                     /annotation_group="60539446776473007ad5a85e-e847-4c38-ba7c
                     -8430819ec8e9"
                     /created_by="primer3"
                     /Sequence="TGTTTGCCGGATCAAGAGCT"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-L1-LHGROATG4-L2"
                     /label="P43"
     misc_feature    4766
                     /Original_Bases="C"
                     /label="C"
                     /note="Geneious type: Editing History Replacement"
     primer_bind     5520..5539
                     /%GC="55.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="60.0"
                     /annotation_group="605394484991230020d1c136-7c0b-4a6b-a758
                     -f16fba6133cd"
                     /created_by="primer3"
                     /Sequence="CTTCCGACTGAGCCTTTCGT"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-L1-LHGROATG4-L2"
                     /label="P46"
ORIGIN      
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgatgagcaa
       61 tgctttttta taatgccaac tttgtacaaa aaagcaggct taatggctag tgaagctcga
      121 aaaacaaaga aaaaaatcaa agggattcag caagccactg caggagtctc acaagacact
      181 tcggaaaatc ctaacaaaac aatagttcct gcagcattac cacagctcac ccctaccttg
      241 gtgtcactgc tggaggtgat tgaacccgag gtgttgtatg caggatatga tagctctgtt
      301 ccagattcag catggagaat tatgaccaca ctcaacatgt taggtgggcg tcaagtgatt
      361 gcagcagtga aatgggcaaa ggcgatacca ggcttcagaa acttacacct ggatgaccaa
      421 atgaccctgc tacagtactc atggatgttt ctcatggcat ttgccctggg ttggagatca
      481 tacagacaat caagtggaaa cctgctctgc tttgctcctg atctgattat taatgagcag
      541 agaatgtctc taccctgcat gtatgaccaa tgtaaacaca tgctgtttgt ctcctctgaa
      601 ttacaaagat tgcaggtatc ctatgaagag tatctctgta tgaaaacctt actgcttctc
      661 tcctcagttc ctaaggaagg tctgaagagc caagagttat ttgatgagat tcgaatgact
      721 tatatcaaag agctaggaaa agccatcgtc aaaagggaag ggaactccag tcagaactgg
      781 caacggtttt accaactgac aaagcttctg gactccatgc atgaggtggt tgagaatctc
      841 cttacctact gcttccagac atttttggat aagaccatga gtattgaatt cccagagatg
      901 ttagctgaaa tcatcactaa tcagatacca aaatattcaa atggaaatat caaaaagctt
      961 ctgtttcatc aaaaatctac tagcaaaccg gtaacgttat acgacgtcgc tgaatacgcc
     1021 ggcgtttctc atcaaaccgt ttctagagtg gttaaccagg cttcacatgt tagcgctaaa
     1081 acccgggaaa aagttgaagc tgccatggct gagctcaact acatcccgaa ccgtgttgcg
     1141 cagcagctgg ctggtaaaca aagcttgctg atcggtgtcg cgacctcgag cttggccctg
     1201 cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc
     1261 gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat
     1321 cttctcgcgc aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc
     1381 attgctgtgg aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag
     1441 acacccatca acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat
     1501 ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg
     1561 gcgcgtctgc gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata
     1621 gcggaacggg aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg
     1681 aatgagggca tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca
     1741 atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt agtgggatac
     1801 gacgataccg aagacagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt
     1861 cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg
     1921 aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa aaaccactag tggatcggaa
     1981 ttcgctaact tcaaccagtc cggaaacatc gctgattctt ccttgagctt cactttcact
     2041 aactcttcta acggacctaa ccttatcacc actcagacca actctcaggc tcttagccag
     2101 ccaatcgcta gctctaacgt gcacgacaac ttcatgaaca acgagatcac tgctagcaag
     2161 atcgatgatg gtaacaattc taagcctctt agcccaggat ggactgatca gactgcttac
     2221 aacgcattcg gtatcactac cggtatgttc aacaccacta ccatggacga tgtgtacaac
     2281 tacctcttcg acgatgagga tactccacct aaccctaaga aggagtgata cccagctttc
     2341 ttgtacaaag ttggcattat aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc
     2401 actatcagtc aaaataaaat cattatttgc catccagctg cagctctggc ccgtgtctca
     2461 aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc
     2521 tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag
     2581 gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa
     2641 tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt
     2701 gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact
     2761 aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga
     2821 tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga
     2881 atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca
     2941 ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc
     3001 gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg
     3061 ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc
     3121 agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattagg
     3181 ccgggaagcc gatctcggct tgaacgaatt gttaggtggc ggtacttggg tcgatatcaa
     3241 agtgcatcac ttcttcccgt atgcccaact ttgtatagag agccactgcg ggatcgtcac
     3301 cgtaatctgc ttgcacgtag atcacataag caccaagcgc gttggcctca tgcttgagga
     3361 gattgatgag cgcggtggca atgccctgcc tccggtgctc gccggagact gcgagatcat
     3421 agatatagat ctcactacgc ggctgctcaa acttgggcag aacgtaagcc gcgagagcgc
     3481 caacaaccgc ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta cggagcaagt
     3541 tcccgaggta atcggagtcc ggctgatgtt gggagtaggt ggctacgtct ccgaactcac
     3601 gaccgaaaag atcaagagca gcccgcatgg atttgacttg gtcagggccg agcctacatg
     3661 tgcgaatgat gcccatactt gagccaccta actttgtttt agggcgactg ccctgctgcg
     3721 taacatcgtt gctgctgcgt aacatcgttg ctgctccata acatcaaaca tcgacccacg
     3781 gcgtaacgcg cttgctgctt ggatgcccga ggcatagact gtacaaaaaa acagtcataa
     3841 caagccatga aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa ggttctggac
     3901 cagttgcgtg agcgcatacg ctacttgcat tacagtttac gaaccgaaca ggcttatgtc
     3961 aactgggttc gtgccttcat ccgtttccac ggtgtgcgtc acccggcaac cttgggcagc
     4021 agcgaagtcg aggcatttct gtcctggctg gcgaacgagc gcaaggtttc ggtctccacg
     4081 catcgtcagg cattggcggc cttgctgttc ttctacggca aggtgctgtg cacggatctg
     4141 ccctggcttc aggagatcgg aagacctcgg ccgtcgcggc gcttgccggt ggtgctgacc
     4201 ccggatgaag tggttcgcat cctcggtttt ctggaaggcg agcatcgttt gttcgcccag
     4261 gactctagct atagttctag tggttggcta ctaataggtt gtattgatgt tggacgagtc
     4321 ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct
     4381 ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa
     4441 ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa
     4501 cactggcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc aaaatccctt
     4561 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     4621 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     4681 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     4741 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     4801 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     4861 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     4921 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     4981 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5041 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5101 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5161 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     5221 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     5281 tatcccctga ttctgtggat aaccgtatta ccgctagcca ggaagagttt gtagaaacgc
     5341 aaaaaggcca tccgtcagga tggccttctg cttagtttga tgcctggcag tttatggcgg
     5401 gcgtcctgcc cgccaccctc cgggccgttg cttcacaacg ttcaaatccg ctcccggcgg
     5461 atttgtccta ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc
     5521 ttccgactga gcctttcgtt ttatttgatg cctggcagtt ccctactctc gcgttaacgc
     5581 tagcatggat ctcgggccc
//
