LOCUS       pEN-L1-R-L2             3222 bp    DNA     circular     06-SEP-2022
DEFINITION  .
ACCESSION   urn.local...hc-dt3ck2q
KEYWORDS    .
SOURCE      
  ORGANISM  .
COMMENT     This file is created by Vector NTI                                 
            http://www.invitrogen.com/                                         
            VNTDATE|459370838|                                                 
            VNTDBDATE|459370838|                                               
            LSOWNER|                                                           
            VNTNAME|pEN-L1-R-L2,0|                                             
            VNTAUTHORNAME|Demo User|                                           
            VNTAUTHOREML|manka@psb.ugent.be|                                   
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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    48
                     /Original_Bases="C"
                     /label="C"
                     /note="Geneious type: Editing History Replacement"
     CDS             103..777
                     /label="RFP"
     primer_bind     332..351
                     /%GC="60.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="60.0"
                     /annotation_group="60520735952078003575887c-74cd-407d-b185
                     -fb4c38c3315f"
                     /created_by="primer3"
                     /Sequence="CCGACATCCCCGACTACATG"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-L1-R-L2"
                     /label="P49"
     attL2           complement(779..878)
                     /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>"
     primer          complement(916..936)
                     /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"
     misc_feature    989..990
                     /Original_Bases="TA"
                     /label="TA"
                     /note="Geneious type: Editing History Replacement"
     promoter        997..1047
                     /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    1048..1857
                     /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     1073..1092
                     /%GC="55.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="25.9"
                     /Tm="59.8"
                     /annotation_group="60520734150058009c4bb0ee-3d88-458f-80b1
                     -5d206a5ec29a"
                     /created_by="primer3"
                     /Sequence="CGAGGCCGCGATTAAATTCC"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-L1-R-L2"
                     /label="P44"
     primer_bind     1707..1726
                     /%GC="60.0"
                     /Hairpin_Tm="40.0"
                     /Self_Dimer_Tm="None"
                     /Tm="60.0"
                     /annotation_group="6052073298502800595170cd-1e3b-4425-9f99
                     -a889c7085b59"
                     /created_by="primer3"
                     /Sequence="CGCAGACCGATACCAGGATC"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-L1-R-L2"
                     /label="P42"
     rep_origin      2004..2592
                     /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     2492..2511
                     /%GC="55.0"
                     /Hairpin_Tm="37.3"
                     /Self_Dimer_Tm="None"
                     /Tm="59.7"
                     /annotation_group="6052073475428300912cb506-8421-4e14-bb86
                     -e98443412427"
                     /created_by="primer3"
                     /Sequence="GGGGAAACGCCTGGTATCTT"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-L1-R-L2"
                     /label="P45"
     primer_bind     3100..3119
                     /%GC="55.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="60.0"
                     /annotation_group="6052073533506300febcb41b-10d3-4b36-b1da
                     -6cf824880549"
                     /created_by="primer3"
                     /Sequence="CTTCCGACTGAGCCTTTCGT"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-L1-R-L2"
                     /label="P46"
     primer          3191..3211
                     /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 tttgtacaaa aaagcaggct taatggacaa caccgaggac
      121 gtcatcaagg agttcatgca gttcaaggtg cgcatggagg gctccgtgaa cggccactac
      181 ttcgagatcg agggcgaggg cgagggcaag ccctacgagg gcacccagac cgccaagctg
      241 caggtgacca agggcggccc cctgcccttc gcctgggaca tcctgtcccc ccagttccag
      301 tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactacat gaagctgtcc
      361 ttccccgagg gcttcacctg ggagcgctcc atgaacttcg aggacggcgg cgtggtggag
      421 gtgcagcagg actcctccct gcaggacggc accttcatct acaaggtgaa gttcaagggc
      481 gtgaacttcc ccgccgacgg ccccgtaatg cagaagaaga ctgccggctg ggagccctcc
      541 accgagaagc tgtaccccca ggacggcgtg ctgaagggcg agatctccca cgccctgaag
      601 ctgaaggacg gcggccacta cacctgcgac ttcaagaccg tgtacaaggc caagaagccc
      661 gtgcagctgc ccggcaacca ctacgtggac tccaagctgg acatcaccaa ccacaacgag
      721 gactacaccg tggtggagca gtacgagcac gccgaggccc gccactccgg ctcccagtac
      781 ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat ttgttgcaac
      841 gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga tatcccctat
      901 agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt gtctcaaaat
      961 ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa actgtctgct
     1021 tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac gtcgaggccg
     1081 cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc
     1141 gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt
     1201 ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac
     1261 tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat
     1321 gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt agaagaatat
     1381 cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg
     1441 attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa
     1501 tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg
     1561 cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc
     1621 gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt
     1681 tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg
     1741 aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt
     1801 gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaatca
     1861 gaattggtta attggttgta acactggcag agcattacgc tgacttgacg ggacggcgca
     1921 agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc
     1981 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     2041 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     2101 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     2161 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     2221 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     2281 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     2341 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     2401 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     2461 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     2521 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     2581 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     2641 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     2701 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     2761 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     2821 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     2881 aatacgcgta ccgctagcca ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga
     2941 tggccttctg cttagtttga tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc
     3001 cgggccgttg cttcacaacg ttcaaatccg ctcccggcgg atttgtccta ctcaggagag
     3061 cgttcaccga caaacaacag ataaaacgaa aggcccagtc ttccgactga gcctttcgtt
     3121 ttatttgatg cctggcagtt ccctactctc gcgttaacgc tagcatggat gttttcccag
     3181 tcacgacgtt gtaaaacgac ggccagtctt aagctcgggc cc
//
