LOCUS       pEN-L1-OL+-L2           4291 bp    DNA     circular     06-SEP-2022
DEFINITION  .
ACCESSION   urn.local...v0-dt4k7ck
KEYWORDS    .
SOURCE      
  ORGANISM  .
COMMENT     This file is created by Vector NTI                                 
            http://www.invitrogen.com/                                         
            VNTDATE|459207056|                                                 
            VNTDBDATE|459207056|                                               
            LSOWNER|                                                           
            VNTNAME|pEn-L1OLplusL2,0|                                          
            VNTAUTHORNAME|Demo User|                                           
            VNTAUTHOREML|manka@psb.ugent.be|                                   
            VNTOAUTHORNAME|Mansour Karimi|                                     
            VNTOAUTHORTEL|+3193313850|                                         
            VNTOAUTHOREML|mansour.karimi@psb.ugent.be|                         
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            -10.4226 0 -10 3.11923 0.694805 #228#))                        
            (CObjectList))                                               
            #230=(CGroupWidget                                                 
                  (CWidget 2 (10 #3# 0) 1 2 0 0 Nil -1842 100)                 
                   (CObjectList                                                
                    #231=(CScratch                                             
                          (CWidget 1 (1 #3# 1) 1 2 0 0 #126# 0 100)            
                               (LOGPEN 0 10 0) 8 1 2.72652 1.9 0.123113 1)     
                                #232=(CLabel                                   
                                      (CWidget 0 (0 0) 1 2 0 0 #154# 27857916
            100)                                (LOGPEN 0 0 8437824) 1         
                                                (LOGFONT 47 17 0 0 400 255 0 0
            0 3 2 1 34                                 "Arial") 1.15807
            0.611111 0                                        "{\\i Ava}I
            (2427)" "@N (@S)" 5 -7.67149                               
            -13.8968 0 -10 3.07488 0.694805 #231#)                         
            #233=(CScratch                                                     
                  (CWidget 2 (1 #3# 2) 1 2 0 0 #126# 0 100)                    
                       (LOGPEN 0 10 0) 8 1 0.883012 1.9 0.123113 1)            
                         #234=(CLabel                                          
                               (CWidget 0 (0 0) 1 2 0 0 #154# 268673032 100)   
                                        (LOGPEN 0 0 8437824) 1                 
                                        (LOGFONT 47 17 0 0 400 255 0 0 0 3 2 1
            34                                 "Arial") 1.15807 0.611111 0     
                                              "{\\i Ava}I (3688)" "@N (@S)" 5
            -12.1208                                8.10649 0 -10 3.07488
            0.694805 #233#))                         (CObjectList)))
            (CObjectList))                         #235=(CGroupWidget (CWidget
            14 (16 0) 1 2 0 0 Nil -947 100)                  (CObjectList)
            (CObjectList))                                  #236=(CGroupWidget
            (CWidget 12 (0 0) 1 2 0 0 Nil -1180 100)                 
            (CObjectList) (CObjectList))) (CObjectList)))         (CSeqView 10
            5 (CObjectList) (CObList) 1 (CObList)) (CObList) 27859216   
            (CStringList) 27859216 0 (CObList))).
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>"
     UTR             101..196
                     /modified_by="mamee"
                     /label="Omega leader"
     CDS             197..1846
                     /label="Luc+"
     primer_bind     669..689
                     /%GC="47.6"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="59.1"
                     /annotation_group="60528379318390005abeb583-6d11-44bf-b00a
                     -ef7af40b16fc"
                     /created_by="primer3"
                     /Sequence="ACCAGGGATTTCAGTCGATGT"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-L1-OL+-L2"
                     /label="P54"
     primer_bind     1470..1489
                     /%GC="55.0"
                     /Hairpin_Tm="42.7"
                     /Self_Dimer_Tm="None"
                     /Tm="60.0"
                     /annotation_group="6052837990930900501c6a7d-c47a-4796-9980
                     -36df15c0a2c9"
                     /created_by="primer3"
                     /Sequence="ACTGGGACGAAGACGAACAC"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-L1-OL+-L2"
                     /label="P55"
     attL2           complement(1848..1947)
                     /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(1985..2005)
                     /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    2058..2059
                     /Original_Bases="TA"
                     /label="TA"
                     /note="Geneious type: Editing History Replacement"
     promoter        2066..2116
                     /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    2117..2926
                     /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     2142..2161
                     /%GC="55.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="25.9"
                     /Tm="59.8"
                     /annotation_group="6052837759069900ad0c569b-b601-4cbc-909d
                     -dfc5c270b5e0"
                     /created_by="primer3"
                     /Sequence="CGAGGCCGCGATTAAATTCC"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-L1-OL+-L2"
                     /label="P44"
     primer_bind     2776..2795
                     /%GC="60.0"
                     /Hairpin_Tm="40.0"
                     /Self_Dimer_Tm="None"
                     /Tm="60.0"
                     /annotation_group="6052837638507000d4f7fdb7-4a9f-4349-b277
                     -fad8aed21773"
                     /created_by="primer3"
                     /Sequence="CGCAGACCGATACCAGGATC"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-L1-OL+-L2"
                     /label="P42"
     rep_origin      3073..3661
                     /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     3561..3580
                     /%GC="55.0"
                     /Hairpin_Tm="37.3"
                     /Self_Dimer_Tm="None"
                     /Tm="59.7"
                     /annotation_group="6052837817119500e3d5cec8-27f7-4e2d-abdd
                     -65388df13cc6"
                     /created_by="primer3"
                     /Sequence="GGGGAAACGCCTGGTATCTT"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-L1-OL+-L2"
                     /label="P45"
     primer_bind     4169..4188
                     /%GC="55.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="60.0"
                     /annotation_group="60528378747830008288105b-ade3-4494-9920
                     -d045278cbfd8"
                     /created_by="primer3"
                     /Sequence="CTTCCGACTGAGCCTTTCGT"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-L1-OL+-L2"
                     /label="P46"
     primer          4260..4280
                     /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 taagatctct gtgggacagc
      121 tttattttta caacaattac caacaacaac aaacaacaaa caacattaca attactattt
      181 acaattacag tcgaccatgg aagacgccaa aaacataaag aaaggcccgg cgccattcta
      241 tccgctggaa gatggaaccg ctggagagca actgcataag gctatgaaga gatacgccct
      301 ggttcctgga acaattgctt ttacagatgc acatatcgag gtggacatca cttacgctga
      361 gtacttcgaa atgtccgttc ggttggcaga agctatgaaa cgatatgggc tgaatacaaa
      421 tcacagaatc gtcgtatgca gtgaaaactc tcttcaattc tttatgccgg tgttgggcgc
      481 gttatttatc ggagttgcag ttgcgcccgc gaacgacatt tataatgaac gtgaattgct
      541 caacagtatg ggcatttcgc agcctaccgt ggtgttcgtt tccaaaaagg ggttgcaaaa
      601 aattttgaac gtgcaaaaaa agctcccaat catccaaaaa attattatca tggattctaa
      661 aacggattac cagggatttc agtcgatgta cacgttcgtc acatctcatc tacctcccgg
      721 ttttaatgaa tacgattttg tgccagagtc cttcgatagg gacaagacaa ttgcactgat
      781 catgaactcc tctggatcta ctggtctgcc taaaggtgtc gctctgcctc atagaactgc
      841 ctgcgtgaga ttctcgcatg ccagagatcc tatttttggc aatcaaatca ttccggatac
      901 tgcgatttta agtgttgttc cattccatca cggttttgga atgtttacta cactcggata
      961 tttgatatgt ggatttcgag tcgtcttaat gtatagattt gaagaagagc tgtttctgag
     1021 gagccttcag gattacaaga ttcaaagtgc gctgctggtg ccaaccctat tctccttctt
     1081 cgccaaaagc actctgattg acaaatacga tttatctaat ttacacgaaa ttgcttctgg
     1141 tggcgctccc ctctctaagg aagtcgggga agcggttgcc aagaggttcc atctgccagg
     1201 tatcaggcaa ggatatgggc tcactgagac tacatcagct attctgatta cacccgaggg
     1261 ggatgataaa ccgggcgcgg tcggtaaagt tgttccattt tttgaagcga aggttgtgga
     1321 tctggatacc gggaaaacgc tgggcgttaa tcaaagaggc gaactgtgtg tgagaggtcc
     1381 tatgattatg tccggttatg taaacaatcc ggaagcgacc aacgccttga ttgacaagga
     1441 tggatggcta cattctggag acatagctta ctgggacgaa gacgaacact tcttcatcgt
     1501 tgaccgcctg aagtctctga ttaagtacaa aggctatcag gtggctcccg ctgaattgga
     1561 atccatcttg ctccaacacc ccaacatctt cgacgcaggt gtcgcaggtc ttcccgacga
     1621 tgacgccggt gaacttcccg ccgccgttgt tgttttggag cacggaaaga cgatgacgga
     1681 aaaagagatc gtggattacg tcgccagtca agtaacaacc gcgaaaaagt tgcgcggagg
     1741 agttgtgttt gtggacgaag taccgaaagg tcttaccgga aaactcgacg caagaaaaat
     1801 cagagagatc ctcataaagg ccaagaaggg cggaaagatc gccgtgtacc cagctttctt
     1861 gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac
     1921 tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta
     1981 ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta
     2041 cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag
     2101 taatacaagg ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc
     2161 caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg
     2221 tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg
     2281 caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga
     2341 atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact
     2401 caccactgcg atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg
     2461 tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg
     2521 taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa
     2581 taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca
     2641 agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg
     2701 tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt
     2761 tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg
     2821 tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga
     2881 tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa
     2941 ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc
     3001 aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat
     3061 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3121 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3181 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     3241 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3301 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3361 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3421 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     3481 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     3541 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     3601 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     3661 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     3721 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     3781 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     3841 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     3901 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atacgcgtac
     3961 cgctagccag gaagagtttg tagaaacgca aaaaggccat ccgtcaggat ggccttctgc
     4021 ttagtttgat gcctggcagt ttatggcggg cgtcctgccc gccaccctcc gggccgttgc
     4081 ttcacaacgt tcaaatccgc tcccggcgga tttgtcctac tcaggagagc gttcaccgac
     4141 aaacaacaga taaaacgaaa ggcccagtct tccgactgag cctttcgttt tatttgatgc
     4201 ctggcagttc cctactctcg cgttaacgct agcatggatg ttttcccagt cacgacgttg
     4261 taaaacgacg gccagtctta agctcgggcc c
//
