LOCUS pEN-R2-6-L3 2910 bp DNA circular 06-SEP-2022 DEFINITION . ACCESSION urn.local...y0-dt4nhn9 KEYWORDS . SOURCE ORGANISM . COMMENT This file is created by Vector NTI http://www.invitrogen.com/ ORIGDB|GenBank VNTDATE|459271037| VNTDBDATE|459271170| LSOWNER| VNTNAME|pEN-R2-6-L3,0| VNTAUTHORNAME|Demo User| Vector_NTI_Display_Data_(Do_Not_Edit!) 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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; attR2" misc_feature 1^2 /Original_Bases="CATAGTGACTGCATATGTTGTGTTTTACAGTATTATGTAGT CTGTTTTTTATGCAAAATCTAATTTAATATATTGATATTTATATCATTTTACGTTTCT CGTTCAACTTTCTTGTACAAAGT" /label="CATAGTGACTGCATATGTTGTGTTTTACAGTATTATGTAGTCTGTTTTTT ATGCAAAATCTAATTTAATATATTGATATTTATATCATTTTACGTTTCTCGTTCAACT TTCTTGTACAAAGT" /note="Geneious type: Editing History Deletion" misc_feature 126..127 /Original_Bases /label /note="Geneious type: Editing History Insertion" terminator 128..395 /note="Derived using Geneious Prime 2020.2.2 'Annotate from Database' based on nucleotide similarity" /X-Transferred_From="NOST" /X-Transferred_Similarity="100.00%" /X-Primary_Match="100.00%; Adjusted-Interval-0:0/0; NOST" /modified_by="mamee" /note="Derived using Geneious Prime 2022.1.1 'Annotate from Database' based on nucleotide similarity" /Transferred_From="tNOS" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; tNOS" /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; tNOS (Basta)" /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; tNOS (Hygromycin)" /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; tNOS (Kanamycin)" /label="tNOS" misc_feature 396 /Original_Bases /label /note="Geneious type: Editing History Insertion" attL3 complement(397..492) /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; attL3" primer_bind 517..536 /%GC="60.0" /Hairpin_Tm="None" /Self_Dimer_Tm="8.2" /Tm="59.3" /annotation_group="60530939983467004922f7d0-cf84-4889-a03b -f12a7be475fc" /created_by="primer3" /Sequence="GGCCGCGATATCCCCTATAG" /Mismatches=0 /#_Off-target_Sites=0 /Off-target_Template="pEN-R2-6-L3" /label="P40" primer complement(550..570) /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 631..681 /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 (pCG034_pKan_Kan-CBE)" /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0; Kan_promoter (pCG040_ABE-Nuc-NLS_Kan -reporter_control)" /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0; Kan_promoter (pCG043_Kan-ABE-Dest_co ntrol_ccdB)" /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0; Kan_promoter (pCG046_ABE-LbCas12a-NL S_Kan_control)" /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0; Kan_promoter (pCG005_pGGD-pKan-KanR- E)" /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0; Kan_promoter (pCG006_pGGA-Kan_promot er-B)" /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0; Kan_promoter (pCG035_pKan_Kan)" /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0; Kan_promoter (pCG039_ABE-N7-NLS_Kan- reporter_control)" /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0; Kan_promoter (pCG041_ABE-SV40-NLS_Ka n-reporter_control)" /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0; Kan_promoter (pCG054_Kan_CBE_control )" /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 (KanR)" /label="kan" misc_feature 682..1491 /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; KanR" /label="KanR" /note="Geneious type: Bacterial Selection" primer_bind 1341..1360 /%GC="60.0" /Hairpin_Tm="40.0" /Self_Dimer_Tm="None" /Tm="60.0" /annotation_group="6053094056750300bf36c071-ebbb-4465-8057 -e9fb44ffbcfe" /created_by="primer3" /Sequence="CGCAGACCGATACCAGGATC" /Mismatches=0 /#_Off-target_Sites=0 /Off-target_Template="pEN-R2-6-L3" /label="P42" rep_origin 1638..2226 /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 (high-copy-number ColE1/pMB1/pBR322/pUC origin of replication)" /label="ColE1/pMB1/pBR322/pUC Ori" misc_feature 1785 /Original_Bases="C" /label="C" /note="Geneious type: Editing History Replacement" misc_feature 2030..2031 /Original_Bases="AT" /label="AT" /note="Geneious type: Editing History Replacement" primer_bind 2126..2145 /%GC="55.0" /Hairpin_Tm="37.3" /Self_Dimer_Tm="None" /Tm="59.7" /annotation_group="60530942332018008bdd4e0e-1e8b-44bf-9c98 -fb4ccc4f1672" /created_by="primer3" /Sequence="GGGGAAACGCCTGGTATCTT" /Mismatches=0 /#_Off-target_Sites=0 /Off-target_Template="pEN-R2-6-L3" /label="P45" primer_bind 2734..2753 /%GC="55.0" /Hairpin_Tm="None" /Self_Dimer_Tm="None" /Tm="60.0" /annotation_group="6053094292412500920fb3d6-4cce-42db-9777 -8b18905af11c" /created_by="primer3" /Sequence="CTTCCGACTGAGCCTTTCGT" /Mismatches=0 /#_Off-target_Sites=0 /Off-target_Template="pEN-R2-6-L3" /label="P46" primer 2825..2845 /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 2909 /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 agtgggacta gagtcaagca gatcgttcaa acatttggca ataaagtttc ttaagattga 181 atcctgttgc cggtcttgcg atgattatca tataatttct gttgaattac gttaagcatg 241 taataattaa catgtaatgc atgacgttat ttatgagatg ggtttttatg attagagtcc 301 cgcaattata catttaatac gcgatagaaa acaaaatata gcgcgcaaac taggataaat 361 tatcgcgcgc ggtgtcatct atgttactag atcgaacaac tttattatac aaagttggca 421 ttataaaaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata 481 aaatcattat ttggagctcc atggtagcgt taacgcggcc gcgatatccc ctatagtgag 541 tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca aaatctctga 601 tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata 661 aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag gccgcgatta 721 aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa 781 tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa 841 catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg 901 acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg 961 ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga atatcctgat 1021 tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct 1081 gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga 1141 atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt 1201 gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact 1261 catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt 1321 gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc 1381 ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat 1441 cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg 1501 gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg cgcaagctca 1561 tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa 1621 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca 1681 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt 1741 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg 1801 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc 1861 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga 1921 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc 1981 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc 2041 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca 2101 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg 2161 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta 2221 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct 2281 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag 2341 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa 2401 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc 2461 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatacg 2521 cgtaccgcta gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct 2581 tctgcttagt ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc 2641 gttgcttcac aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca 2701 ccgacaaaca acagataaaa cgaaaggccc agtcttccga ctgagccttt cgttttattt 2761 gatgcctggc agttccctac tctcgcgtta acgctagcat ggatgttttc ccagtcacga 2821 cgttgtaaaa cgacggccag tcttaagctc gggccctgca gctctagagc tcgaattcta 2881 caggtcacta ataccatcta agtagttggt //