LOCUS       pEN-R2-F-L3             3359 bp    DNA     circular     06-SEP-2022
DEFINITION  .
ACCESSION   urn.local...m4-dt3f258
KEYWORDS    .
SOURCE      
  ORGANISM  .
COMMENT     This file is created by Vector NTI                                 
            http://www.invitrogen.com/                                         
            VNTDATE|458702925|                                                 
            VNTDBDATE|458702925|                                               
            LSOWNER|                                                           
            VNTNAME|pEN-R2FL3,0|                                               
            VNTAUTHORNAME|Demo User|                                           
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            1) 1 2 0 0 #177# 0 100)                                (LOGPEN 0 0
            10079283) 1 1 1.67895 1.9                                0.123113
            1)                                                   #233=(CLabel  
                                                                       (CWidget
            0 (0 0) 1 2 0 0 #157# 393240 100)                               
            (LOGPEN 0 0 8437824) 1                                             
            (LOGFONT 39 14 0 0 400 255 1 0 0 3 2 1 34                          
                  "Arial") 1.15807 0.611111 0                                  
                 "{\\i Xba}I (2461)" "@N (@S)" 5 -14.4526                      
                     -1.15807 0 -10 2.91758 0.711604 #232#))                   
                 (CObjectList))                                               
            #234=(CGroupWidget                                                 
                  (CWidget 2 (10 #14# 0) 1 2 0 0 Nil -372 100)                 
                   (CObjectList                                                
                    #235=(CScratch                                             
                          (CWidget 1 (1 #14# 1) 1 2 0 0 #177# 0 100)           
                                (LOGPEN 0 0 10079283) 1 1 6.09577 1.9          
                                 0.123113 1)                                   
                           #236=(CLabel                                        
                                 (CWidget 0 (0 0) 1 2 0 0 #156# 4129304 100)   
                                        (LOGPEN 0 0 8437824) 1                 
                                        (LOGFONT 39 14 0 0 400 255 0 0 0 3 2 1
            34                                 "Arial") 1.15807 0.611111 0     
                                              "{\\i Sac}I (96)" "@N (@S)" 5
            4.46165 13.8968                                0 -10 2.31271
            0.711604 #235#)                                 #237=(CScratch     
                                                                  (CWidget 2 (1
            #14# 2) 1 2 0 0 #177# 0 100)                                (LOGPEN
            0 0 10079283) 1 1 1.66401 1.9                               
            0.123113 1)                                                  
            #238=(CLabel                                                       
                  (CWidget 0 (0 0) 1 2 0 0 #156# 4129304 100)                  
                         (LOGPEN 0 0 8437824) 1                                
                         (LOGFONT 39 14 0 0 400 255 0 0 0 3 2 1 34             
                               "Arial") 1.15807 0.611111 0                     
                              "{\\i Sac}I (2469)" "@N (@S)" 5 -14.4949 0 0     
                                      -10 2.91758 0.711604 #237#))
            (CObjectList)))                  (CObjectList))                    
                                       #239=(CGroupWidget (CWidget 14 (16 0) 1
            2 0 0 Nil -930 100)                  (CObjectList) (CObjectList))  
                                           #240=(CGroupWidget (CWidget 12 (0 0)
            1 2 0 0 Nil -1622 100)                  (CObjectList)
            (CObjectList))) (CObjectList)))         (CSeqView 10 5
            (CObjectList) (CObList) 1 (CObList)) (CObList) 40232704   
            (CStringList) 0 128 (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..844
                     /label="GFP"
     primer_bind     411..430
                     /%GC="55.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="26.0"
                     /Tm="60.1"
                     /annotation_group="60521906740413008ab80846-14f3-4597-908f
                     -9058c453b425"
                     /created_by="primer3"
                     /Sequence="AGGAGCGCACCATCTTCTTC"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-R2-F-L3"
                     /label="P47"
     attL3           complement(846..941)
                     /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(999..1019)
                     /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        1080..1130
                     /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    1131..1940
                     /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     1156..1175
                     /%GC="55.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="25.9"
                     /Tm="59.8"
                     /annotation_group="6052190498802500586cb85c-1d5d-4ef8-b327
                     -6e9e7a6b0208"
                     /created_by="primer3"
                     /Sequence="CGAGGCCGCGATTAAATTCC"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-R2-F-L3"
                     /label="P44"
     primer_bind     1790..1809
                     /%GC="60.0"
                     /Hairpin_Tm="40.0"
                     /Self_Dimer_Tm="None"
                     /Tm="60.0"
                     /annotation_group="60521903711995002af19cc2-600a-4226-882f
                     -954a3fd39a71"
                     /created_by="primer3"
                     /Sequence="CGCAGACCGATACCAGGATC"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-R2-F-L3"
                     /label="P42"
     rep_origin      2087..2675
                     /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    2234
                     /Original_Bases="C"
                     /label="C"
                     /note="Geneious type: Editing History Replacement"
     misc_feature    2479..2480
                     /Original_Bases="AT"
                     /label="AT"
                     /note="Geneious type: Editing History Replacement"
     primer_bind     2575..2594
                     /%GC="55.0"
                     /Hairpin_Tm="37.3"
                     /Self_Dimer_Tm="None"
                     /Tm="59.7"
                     /annotation_group="6052190558570100ce02c7ac-a503-43c3-bd8a
                     -0e18519c8417"
                     /created_by="primer3"
                     /Sequence="GGGGAAACGCCTGGTATCTT"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-R2-F-L3"
                     /label="P45"
     primer_bind     3183..3202
                     /%GC="55.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="60.0"
                     /annotation_group="60521906163208007643de5e-1f38-4186-8872
                     -5fd9310a7201"
                     /created_by="primer3"
                     /Sequence="CTTCCGACTGAGCCTTTCGT"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pEN-R2-F-L3"
                     /label="P46"
     primer          3274..3294
                     /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    3358
                     /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 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
      181 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
      241 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
      301 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
      361 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
      421 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
      481 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
      541 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
      601 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
      661 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
      721 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
      781 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
      841 caagacaact ttattataca aagttggcat tataaaaaag cattgcttat caatttgttg
      901 caacgaacag gtcactatca gtcaaaataa aatcattatt tggagctcca tggtagcgtt
      961 aacgcggccg cgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc
     1021 agctctggcc cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat
     1081 catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata
     1141 ttcaacggga aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt
     1201 ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga
     1261 agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta
     1321 cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc
     1381 attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag
     1441 cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag
     1501 tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg
     1561 tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt
     1621 ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt
     1681 tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt
     1741 ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat
     1801 accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac
     1861 ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga
     1921 tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta
     1981 cgctgacttg acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt
     2041 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     2101 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     2161 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     2221 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     2281 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     2341 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     2401 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     2461 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     2521 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     2581 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     2641 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     2701 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     2761 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     2821 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     2881 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     2941 cgggcagtga gcgcaacgca attaatacgc gtaccgctag ccaggaagag tttgtagaaa
     3001 cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg
     3061 cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg
     3121 cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca
     3181 gtcttccgac tgagcctttc gttttatttg atgcctggca gttccctact ctcgcgttaa
     3241 cgctagcatg gatgttttcc cagtcacgac gttgtaaaac gacggccagt cttaagctcg
     3301 ggccctgcag ctctagagct cgaattctac aggtcactaa taccatctaa gtagttggt
//
