LOCUS       pEN-R2-L+-L3            4295 bp    DNA     circular     06-SEP-2022
DEFINITION  .
ACCESSION   urn.local...ws-dt4ly1t
KEYWORDS    .
SOURCE      
  ORGANISM  .
COMMENT     This file is created by Vector NTI                                 
            http://www.invitrogen.com/                                         
            VNTDATE|459274933|                                                 
            VNTDBDATE|459274933|                                               
            LSOWNER|                                                           
            VNTNAME|pEN-R2-L+-L3,0|                                            
            VNTAUTHORNAME|Demo User|                                           
            Vector_NTI_Display_Data_(Do_Not_Edit!)                             
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            "HindIII" "aagctt" 2 0 0 0 "")             #9=(CRSite (CStringList)
            "NcoI" "ccatgg" 2 0 1 97 0 0 "")           #10=(CRSite
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            #15=(CRSite (CStringList) "SacII" "ccgcgg" 5 0 0 0 "")             
            #16=(CRSite (CStringList) "SalI" "gtcgac" 2 0 0 0 "")              
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                                                          (CWidget 2 (1 #3# 2)
            1 2 0 0 #178# 4129224                                 100) (LOGPEN
            0 0 10079283) 1 1 0.880934 1.9                               
            0.123113 1)                                                  
            #232=(CLabel                                                       
                  (CWidget 0 (0 0) 1 2 0 0 #157# 393220 100)                   
                        (LOGPEN 0 0 8437824) 1                                 
                        (LOGFONT 43 16 0 0 400 255 0 0 0 3 2 1 34              
                              "Arial") 1.15807 0.611111 0                      
                             "{\\i Ava}I (3692)" "@N (@S)" 5 -12.0339          
                                 8.10649 0 -10 2.96138 0.68832 #231#))         
                           (CObjectList))                                      
                     #233=(CGroupWidget                                        
                           (CWidget 2 (10 #14# 0) 1 2 0 0 Nil -110 100)        
                            (CObjectList                                       
                             #234=(CScratch                                    
                                   (CWidget 1 (1 #14# 1) 1 2 0 0 #178# -187
            100)                                (LOGPEN 0 0 10079283) 1 1
            6.1344 1.9 0.123113                                1)              
                                                         #235=(CLabel          
                                                               (CWidget 0 (0 0)
            1 2 0 0 #157# 1953789025 100)                               
            (LOGPEN 0 0 8437824) 1                                             
            (LOGFONT 43 16 0 0 400 255 0 0 0 3 2 1 34                          
                  "Arial") 1.15807 0.611111 0                                  
                 "{\\i Sac}I (96)" "@N (@S)" 5 3.96826 13.8968                 
                          0 -10 2.32108 0.68832 #234#)                         
                    #236=(CScratch                                             
                          (CWidget 2 (1 #14# 2) 1 2 0 0 #178# 4129224          
                                  100) (LOGPEN 0 0 10079283) 1 1 2.66764 1.9   
                                        0.123113 1)                            
                                  #237=(CLabel                                 
                                        (CWidget 0 (0 0) 1 2 0 0 #157# 27844224
            100)                                (LOGPEN 0 0 8437824) 1         
                                                (LOGFONT 43 16 0 0 400 255 0 0
            0 3 2 1 34                                 "Arial") 1.15807
            0.611111 0                                        "{\\i Sac}I
            (2469)" "@N (@S)" 5 -9.80578                               
            -10.4226 0 -10 2.96138 0.68832 #236#))                        
            (CObjectList))) (CObjectList))                        
            #238=(CGroupWidget (CWidget 14 (16 0) 1 2 0 0 Nil -1011 100)       
                      (CObjectList) (CObjectList))                             
                #239=(CGroupWidget (CWidget 12 (0 0) 1 2 0 0 Nil 0 100)        
                      (CObjectList) (CObjectList))) (CObjectList)))        
            (CSeqView 10 5 (CObjectList) (CObList) 1 (CObList)) (CObList)
            50855986    (CStringList) 1074193686 327685 (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..1780
                     /label="LUC+"
     primer_bind     600..620
                     /%GC="47.6"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="59.1"
                     /annotation_group="6052991592625500a513b86a-4375-4de9-a81f
                     -bc5588309abf"
                     /created_by="primer3"
                     /Sequence="ACCAGGGATTTCAGTCGATGT"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-R2-L+-L3"
                     /label="P54"
     primer_bind     1401..1420
                     /%GC="55.0"
                     /Hairpin_Tm="42.7"
                     /Self_Dimer_Tm="None"
                     /Tm="60.0"
                     /annotation_group="6052991649568300fd1096da-4b26-4214-9046
                     -cdef4b845874"
                     /created_by="primer3"
                     /Sequence="ACTGGGACGAAGACGAACAC"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-R2-L+-L3"
                     /label="P55"
     misc_feature    1781
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     attL3           complement(1782..1877)
                     /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          complement(1935..1955)
                     /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"
     promoter        2016..2066
                     /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    2067..2876
                     /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     2092..2111
                     /%GC="55.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="25.9"
                     /Tm="59.8"
                     /annotation_group="6052991418236900ae685cdf-68a7-477a-b0e9
                     -2bfa2592985f"
                     /created_by="primer3"
                     /Sequence="CGAGGCCGCGATTAAATTCC"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-R2-L+-L3"
                     /label="P44"
     primer_bind     2726..2745
                     /%GC="60.0"
                     /Hairpin_Tm="40.0"
                     /Self_Dimer_Tm="None"
                     /Tm="60.0"
                     /annotation_group="60529913030522001110bdb9-d778-44c1-b214
                     -2a971cacb151"
                     /created_by="primer3"
                     /Sequence="CGCAGACCGATACCAGGATC"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-R2-L+-L3"
                     /label="P42"
     rep_origin      3023..3611
                     /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"
     misc_feature    3170
                     /Original_Bases="C"
                     /label="C"
                     /note="Geneious type: Editing History Replacement"
     misc_feature    3415..3416
                     /Original_Bases="AT"
                     /label="AT"
                     /note="Geneious type: Editing History Replacement"
     primer_bind     3511..3530
                     /%GC="55.0"
                     /Hairpin_Tm="37.3"
                     /Self_Dimer_Tm="None"
                     /Tm="59.7"
                     /annotation_group="6052991476024100568cbbad-710e-4a97-b0e3
                     -55917b347602"
                     /created_by="primer3"
                     /Sequence="GGGGAAACGCCTGGTATCTT"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-R2-L+-L3"
                     /label="P45"
     primer_bind     4119..4138
                     /%GC="55.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="60.0"
                     /annotation_group="6052991535101100f054d0b0-91a6-4482-959a
                     -8a82125f37ff"
                     /created_by="primer3"
                     /Sequence="CTTCCGACTGAGCCTTTCGT"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-R2-L+-L3"
                     /label="P46"
     primer          4210..4230
                     /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"
     misc_feature    4294
                     /Original_Bases="A"
                     /label="A"
                     /note="Geneious type: Editing History Replacement"
ORIGIN      
        1 tcatagtgac tgcatatgtt gtgttttaca gtattatgta gtctgttttt tatgcaaaat
       61 ctaatttaat atattgatat ttatatcatt ttacgtttct cgttcaactt tcttgtacaa
      121 agtgggaatg gaagacgcca aaaacataaa gaaaggcccg gcgccattct atccgctgga
      181 agatggaacc gctggagagc aactgcataa ggctatgaag agatacgccc tggttcctgg
      241 aacaattgct tttacagatg cacatatcga ggtggacatc acttacgctg agtacttcga
      301 aatgtccgtt cggttggcag aagctatgaa acgatatggg ctgaatacaa atcacagaat
      361 cgtcgtatgc agtgaaaact ctcttcaatt ctttatgccg gtgttgggcg cgttatttat
      421 cggagttgca gttgcgcccg cgaacgacat ttataatgaa cgtgaattgc tcaacagtat
      481 gggcatttcg cagcctaccg tggtgttcgt ttccaaaaag gggttgcaaa aaattttgaa
      541 cgtgcaaaaa aagctcccaa tcatccaaaa aattattatc atggattcta aaacggatta
      601 ccagggattt cagtcgatgt acacgttcgt cacatctcat ctacctcccg gttttaatga
      661 atacgatttt gtgccagagt ccttcgatag ggacaagaca attgcactga tcatgaactc
      721 ctctggatct actggtctgc ctaaaggtgt cgctctgcct catagaactg cctgcgtgag
      781 attctcgcat gccagagatc ctatttttgg caatcaaatc attccggata ctgcgatttt
      841 aagtgttgtt ccattccatc acggttttgg aatgtttact acactcggat atttgatatg
      901 tggatttcga gtcgtcttaa tgtatagatt tgaagaagag ctgtttctga ggagccttca
      961 ggattacaag attcaaagtg cgctgctggt gccaacccta ttctccttct tcgccaaaag
     1021 cactctgatt gacaaatacg atttatctaa tttacacgaa attgcttctg gtggcgctcc
     1081 cctctctaag gaagtcgggg aagcggttgc caagaggttc catctgccag gtatcaggca
     1141 aggatatggg ctcactgaga ctacatcagc tattctgatt acacccgagg gggatgataa
     1201 accgggcgcg gtcggtaaag ttgttccatt ttttgaagcg aaggttgtgg atctggatac
     1261 cgggaaaacg ctgggcgtta atcaaagagg cgaactgtgt gtgagaggtc ctatgattat
     1321 gtccggttat gtaaacaatc cggaagcgac caacgccttg attgacaagg atggatggct
     1381 acattctgga gacatagctt actgggacga agacgaacac ttcttcatcg ttgaccgcct
     1441 gaagtctctg attaagtaca aaggctatca ggtggctccc gctgaattgg aatccatctt
     1501 gctccaacac cccaacatct tcgacgcagg tgtcgcaggt cttcccgacg atgacgccgg
     1561 tgaacttccc gccgccgttg ttgttttgga gcacggaaag acgatgacgg aaaaagagat
     1621 cgtggattac gtcgccagtc aagtaacaac cgcgaaaaag ttgcgcggag gagttgtgtt
     1681 tgtggacgaa gtaccgaaag gtcttaccgg aaaactcgac gcaagaaaaa tcagagagat
     1741 cctcataaag gccaagaagg gcggaaagat cgccgtgtaa acaactttat tatacaaagt
     1801 tggcattata aaaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca
     1861 aaataaaatc attatttgga gctccatggt agcgttaacg cggccgcgat atcccctata
     1921 gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc
     1981 tctgatgtta cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt
     2041 acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg tcgaggccgc
     2101 gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg
     2161 ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc
     2221 tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact
     2281 ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg
     2341 catggttact caccactgcg atccccggaa aaacagcatt ccaggtatta gaagaatatc
     2401 ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga
     2461 ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat
     2521 cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc
     2581 ctgttgaaca agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg
     2641 tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt
     2701 gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga
     2761 actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg
     2821 ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag
     2881 aattggttaa ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcgcaa
     2941 gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg
     3001 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     3061 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     3121 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     3181 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     3241 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     3301 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     3361 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     3421 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     3481 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     3541 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     3601 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     3661 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     3721 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     3781 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     3841 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     3901 atacgcgtac cgctagccag gaagagtttg tagaaacgca aaaaggccat ccgtcaggat
     3961 ggccttctgc ttagtttgat gcctggcagt ttatggcggg cgtcctgccc gccaccctcc
     4021 gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga tttgtcctac tcaggagagc
     4081 gttcaccgac aaacaacaga taaaacgaaa ggcccagtct tccgactgag cctttcgttt
     4141 tatttgatgc ctggcagttc cctactctcg cgttaacgct agcatggatg ttttcccagt
     4201 cacgacgttg taaaacgacg gccagtctta agctcgggcc ctgcagctct agagctcgaa
     4261 ttctacaggt cactaatacc atctaagtag ttggt
//
