LOCUS pEN-L4-2-R1 3769 bp DNA circular 06-SEP-2022 DEFINITION . ACCESSION urn.local...r3-doze616 KEYWORDS . SOURCE ORGANISM . COMMENT This file is created by Vector NTI http://www.invitrogen.com/ ORIGDB|GenBank VNTDATE|459275533| VNTDBDATE|459275533| LSOWNER| VNTNAME|pEN-L4-2-R1,0| VNTAUTHORNAME|Demo User| Vector_NTI_Display_Data_(Do_Not_Edit!) 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FEATURES Location/Qualifiers attL4 1..97 /Color="#00cccc" /created_by="mamee" /note="Derived using Geneious Prime 2022.1.1 'Annotate from Database' based on nucleotide similarity" /Transferred_From="attL4" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; attL4" misc_feature 98..99 /Original_Bases /label /note="Geneious type: Editing History Insertion" promoter 148..1194 /modified_by="mamee" /note="Derived using Geneious Prime 2022.1.1 'Annotate from Database' based on nucleotide similarity" /Transferred_From="pCaMV 35S - Gateway" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; p35S (pCaMV 35S - Gateway)" /label="p35S" misc_feature 168..180 /Original_Bases /label /note="Geneious type: Editing History Insertion" primer_bind 331..351 /%GC="57.1" /Hairpin_Tm="None" /Self_Dimer_Tm="4.4" /Tm="60.0" /annotation_group="60523053606300000814a7f4-bb8a-49a8-b7fa -d26557e9de2a" /created_by="primer3" /Sequence="GATGGTTAGAGAGGCCTACGC" /Mismatches=0 /#_Off-target_Sites=0 /Off-target_Template="pEN-L4-2-R1" /label="P10" primer_bind 817..836 /%GC="55.0" /Hairpin_Tm="None" /Self_Dimer_Tm="3.1" /Tm="60.2" /annotation_group="60523054200048009c5c0bec-d599-4051-8978 -7b3e8b59d21a" /created_by="primer3" /Sequence="CGGATTCCATTGCCCAGCTA" /Mismatches=0 /#_Off-target_Sites=0 /Off-target_Template="pEN-L4-2-R1" /label="P11" misc_feature 929 /Original_Bases="T" /label="T" /note="Geneious type: Editing History Replacement" misc_feature 1124 /Original_Bases="G" /label="G" /note="Geneious type: Editing History Replacement" misc_feature 1126 /Original_Bases="A" /label="A" /note="Geneious type: Editing History Replacement" misc_feature 1127^1128 /Original_Bases="GACGATCCG" /label="GACGATCCG" /note="Geneious type: Editing History Deletion" misc_feature 1223 /Original_Bases /label /note="Geneious type: Editing History Insertion" attR1 1224..1380 /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="attR1" /Transferred_Similarity="99.36%" /Primary_Match="99.36%; Adjusted-Interval-0:0/0; attR1" misc_feature 1350 /Original_Bases="T" /label="T" /note="Geneious type: Editing History Replacement" primer complement(1443..1459) /invitrogen=2010000 /note="Derived using Geneious Prime 2020.2.2 'Annotate from Database' based on nucleotide similarity" /Transferred_From="pDONRP4-P3 - M13 reverse primer" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; M13 reverse primer (pDONRP4-P3 - M13 reverse primer)" /label="M13 reverse primer" promoter 1521..1571 /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 1572..2381 /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 1597..1616 /%GC="55.0" /Hairpin_Tm="None" /Self_Dimer_Tm="25.9" /Tm="59.8" /annotation_group="60523057819143003871436a-6251-4c58-a35e -437fb617fa5d" /created_by="primer3" /Sequence="CGAGGCCGCGATTAAATTCC" /Mismatches=0 /#_Off-target_Sites=0 /Off-target_Template="pEN-L4-2-R1" /label="P44" primer_bind 2231..2250 /%GC="60.0" /Hairpin_Tm="40.0" /Self_Dimer_Tm="None" /Tm="60.0" /annotation_group="605230565714010024e5cde3-0cd1-4f10-b6fb -3da691dda779" /created_by="primer3" /Sequence="CGCAGACCGATACCAGGATC" /Mismatches=0 /#_Off-target_Sites=0 /Off-target_Template="pEN-L4-2-R1" /label="P42" rep_origin 2528..3116 /created_by="thjac" /modified_by="mamee" /note="Derived using Geneious Prime 2020.2.2 '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 2675 /Original_Bases="C" /label="C" /note="Geneious type: Editing History Replacement" misc_feature 2920..2921 /Original_Bases="AT" /label="AT" /note="Geneious type: Editing History Replacement" primer_bind 3016..3035 /%GC="55.0" /Hairpin_Tm="37.3" /Self_Dimer_Tm="None" /Tm="59.7" /annotation_group="60523058493578006978c923-96f3-4d83-86b8 -72f3eca53f48" /created_by="primer3" /Sequence="GGGGAAACGCCTGGTATCTT" /Mismatches=0 /#_Off-target_Sites=0 /Off-target_Template="pEN-L4-2-R1" /label="P45" primer_bind 3624..3643 /%GC="55.0" /Hairpin_Tm="None" /Self_Dimer_Tm="None" /Tm="60.0" /annotation_group="6052305907507000b5b5359e-3d25-4a5a-980f -5bf90e9a5945" /created_by="primer3" /Sequence="CTTCCGACTGAGCCTTTCGT" /Mismatches=0 /#_Off-target_Sites=0 /Off-target_Template="pEN-L4-2-R1" /label="P46" primer 3695..3711 /invitrogen=2000000 /note="Derived using Geneious Prime 2020.2.2 'Annotate from Database' based on nucleotide similarity" /Transferred_From="pDONRP4-P3 - M13 (-40) forward primer" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; M13 (-40) forward primer (pDONRP4-P3 - M13 (-40) forward primer)" /label="M13 (-40) forward primer" primer 3715..3730 /invitrogen=1980000 /note="Derived using Geneious Prime 2020.2.2 'Annotate from Database' based on nucleotide similarity" /Transferred_From="pDONRP4-P3 - M13 (-20) forward primer" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; M13 (-20) forward primer (pDONRP4-P3 - M13 (-20) forward primer)" /label="M13 (-20) forward primer" misc_feature 3748 /Original_Bases="C" /label="C" /note="Geneious type: Editing History Replacement" ORIGIN 1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgataagcaa 61 tgctttttta taatgccaac tttgtataga aaagttgtca tttaggtgac actatagaat 121 actcaagcta tgcatccaac gcgttgggag ctctcccata tggtcgacta gagccaagct 181 gatctccttt gccccggaga tcaccatgga cgactttctc tatctctacg atctaggaag 241 aaagttcgac ggagaaggtg acgataccat gttcaccacc gataatgaga agattagcct 301 cttcaatttc agaaagaatg ctgacccaca gatggttaga gaggcctacg cggcaggtct 361 catcaagacg atctacccga gtaataatct ccaggagatc aaataccttc ccaagaaggt 421 taaagatgca gtcaaaagat tcaggactaa ctgcatcaag aacacagaga aagatatatt 481 tctcaagatc agaagtacta ttccagtatg gacgattcaa ggcttgcttc ataaaccaag 541 gcaagtaata gagattggag tctctaagaa agtagttcct actgaatcaa aggccatgga 601 gtcaaaaatt cagatcgagg atctaacaga actcgccgtg aagactggcg aacagttcat 661 acagagtctt ttacgactca atgacaagaa gaaaatcttc gtcaacatgg tggagcacga 721 cactctcgtc tactccaaga atatcaaaga tacagtctca gaagaccaaa gggctattga 781 gacttttcaa caaagggtaa tatcgggaaa cctcctcgga ttccattgcc cagctatctg 841 tcacttcatc aaaaggacag tagaaaagga aggtggcacc tacaaatgcc atcattgcga 901 taaaggaaag gctatcgttc aagatgcccc tgccgacagt ggtcccaaag atggaccccc 961 acccacgagg agcatcgtgg aaaaagaaga cgttccaacc acgtcttcaa agcaagtgga 1021 ttgatgtgat atctccactg acgtaaggga tgacgcacaa tcccactatc cttcgcaaga 1081 cccttcctct atataaggaa gttcatttca tttggagagg actccggtat ttttacaaca 1141 attaccacaa caaaacaaac aacaaacaac attacaattt actattctag tcgacctgca 1201 ggcggccgca ctagtgatat cagcaagttt gtacaaaaaa gttgaacgag aaacgtaaaa 1261 tgatataaat atcaatatat taaattagat tttgcataaa aaacagacta cataatactg 1321 taaaacacaa catatgcagt cactatgaac caactactta gatggtatta gtgacctgta 1381 gaattcgagc tctagagctg cagggcggcc gcgatatccc ctatagtgag tcgtattaca 1441 tggtcatagc tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg 1501 cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata 1561 caaggggtgt tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca 1621 tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga 1681 caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag 1741 gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta 1801 tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca 1861 ctgcgatccc cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa 1921 atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt 1981 gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg 2041 gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct 2101 ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt 2161 tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac 2221 gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt 2281 tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga 2341 ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt 2401 tgtaacactg gcagagcatt acgctgactt gacgggacgg cgcaagctca tgaccaaaat 2461 cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag 2521 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac 2581 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa 2641 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc 2701 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag 2761 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac 2821 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc 2881 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc 2941 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca 3001 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc 3061 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg 3121 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct 3181 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata 3241 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc 3301 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg 3361 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatacg cgtaccgcta 3421 gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct tctgcttagt 3481 ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc gttgcttcac 3541 aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca ccgacaaaca 3601 acagataaaa cgaaaggccc agtcttccga ctgagccttt cgttttattt gatgcctggc 3661 agttccctac tctcgcgtta acgctagcat ggatgttttc ccagtcacga cgttgtaaaa 3721 cgacggccag tcttaagctc gggcccgagt taacgctacc atggagctc //