LOCUS pEN-L1-NR-L2 3270 bp DNA circular 06-SEP-2022 DEFINITION . ACCESSION urn.local...ab-dstok26 KEYWORDS . SOURCE ORGANISM . COMMENT This file is created by Vector NTI http://www.invitrogen.com/ VNTDATE|459279823| VNTDBDATE|459279823| LSOWNER| VNTNAME|pEN-L1-NR-L2,0| VNTAUTHORNAME|Demo User| Vector_NTI_Display_Data_(Do_Not_Edit!) 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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; attL1" misc_feature 48 /Original_Bases="C" /label="C" /note="Geneious type: Editing History Replacement" NLS 103..150 /created_by="wacae" /note="Derived using Geneious Prime 2021.2.2 'Annotate from Database' based on nucleotide similarity" /Transferred_From="NLS c2" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; c2 NLS (NLS c2)" /modified_by="mamee" /label="c2 NLS" CDS 151..825 /label="RFP" primer_bind 380..399 /#_Off-target_Sites=0 /Off-target_Template="pEN-L1-NR-L2" /Sequence="CCGACATCCCCGACTACATG" /Tm="60.0" /%GC="60.0" /Hairpin_Tm="None" /Self_Dimer_Tm="None" /modified_by="mamee" /label="P49" attL2 complement(827..926) /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; attL2" primer complement(964..984) /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 1037..1038 /Original_Bases="TA" /label="TA" /note="Geneious type: Editing History Replacement" promoter 1045..1095 /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 1096..1905 /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 1121..1140 /%GC="55.0" /Hairpin_Tm="None" /Self_Dimer_Tm="25.9" /Tm="59.8" /annotation_group="60509420760600006e1a8ca0-a3d9-4376-9311 -ad53ff8bd794" /created_by="primer3" /Sequence="CGAGGCCGCGATTAAATTCC" /Mismatches=0 /#_Off-target_Sites=0 /Off-target_Template="pEN-L1-NR-L2" /label="P44" primer_bind 1755..1774 /%GC="60.0" /Hairpin_Tm="40.0" /Self_Dimer_Tm="None" /Tm="60.0" /annotation_group="6050941950798400c18cf616-0a9c-46ec-98d7 -284ff3b36e67" /created_by="primer3" /Sequence="CGCAGACCGATACCAGGATC" /Mismatches=0 /#_Off-target_Sites=0 /Off-target_Template="pEN-L1-NR-L2" /label="P42" rep_origin 2052..2640 /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" primer_bind 2540..2559 /%GC="55.0" /Hairpin_Tm="37.3" /Self_Dimer_Tm="None" /Tm="59.7" /annotation_group="6050942138333400e3650117-daae-4e0b-9233 -aac7147a32ec" /created_by="primer3" /Sequence="GGGGAAACGCCTGGTATCTT" /Mismatches=0 /#_Off-target_Sites=0 /Off-target_Template="pEN-L1-NR-L2" /label="P45" primer_bind 3148..3167 /%GC="55.0" /Hairpin_Tm="None" /Self_Dimer_Tm="None" /Tm="60.0" /annotation_group="6050942198119400ed243500-4496-400d-9fab -1f7eb79b883c" /created_by="primer3" /Sequence="CTTCCGACTGAGCCTTTCGT" /Mismatches=0 /#_Off-target_Sites=0 /Off-target_Template="pEN-L1-NR-L2" /label="P46" primer 3239..3259 /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 ccatgcagcc ttctcttaaa 121 cgcatgaaaa tagagccatc ttctcaacct atggacaaca ccgaggacgt catcaaggag 181 ttcatgcagt tcaaggtgcg catggagggc tccgtgaacg gccactactt cgagatcgag 241 ggcgagggcg agggcaagcc ctacgagggc acccagaccg ccaagctgca ggtgaccaag 301 ggcggccccc tgcccttcgc ctgggacatc ctgtcccccc agttccagta cggctccaag 361 gcctacgtga agcaccccgc cgacatcccc gactacatga agctgtcctt ccccgagggc 421 ttcacctggg agcgctccat gaacttcgag gacggcggcg tggtggaggt gcagcaggac 481 tcctccctgc aggacggcac cttcatctac aaggtgaagt tcaagggcgt gaacttcccc 541 gccgacggcc ccgtaatgca gaagaagact gccggctggg agccctccac cgagaagctg 601 tacccccagg acggcgtgct gaagggcgag atctcccacg ccctgaagct gaaggacggc 661 ggccactaca cctgcgactt caagaccgtg tacaaggcca agaagcccgt gcagctgccc 721 ggcaaccact acgtggactc caagctggac atcaccaacc acaacgagga ctacaccgtg 781 gtggagcagt acgagcacgc cgaggcccgc cactccggct cccagtaccc agctttcttg 841 tacaaagttg gcattataag aaagcattgc ttatcaattt gttgcaacga acaggtcact 901 atcagtcaaa ataaaatcat tatttgccat ccagctgata tcccctatag tgagtcgtat 961 tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac 1021 attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt 1081 aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg attaaattcc 1141 aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt 1201 gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct gaaacatggc 1261 aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa 1321 tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc 1381 accactgcga tccccggaaa aacagcattc caggtattag aagaatatcc tgattcaggt 1441 gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt 1501 aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat 1561 aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa 1621 gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt 1681 gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt 1741 ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt 1801 gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat 1861 atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga attggttaat 1921 tggttgtaac actggcagag cattacgctg acttgacggg acggcgcaag ctcatgacca 1981 aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc 2041 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa 2101 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag 2161 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta 2221 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta 2281 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag 2341 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg 2401 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg 2461 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag 2521 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc 2581 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa 2641 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg 2701 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct 2761 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa 2821 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg 2881 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc 2941 gctagccagg aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg gccttctgct 3001 tagtttgatg cctggcagtt tatggcgggc gtcctgcccg ccaccctccg ggccgttgct 3061 tcacaacgtt caaatccgct cccggcggat ttgtcctact caggagagcg ttcaccgaca 3121 aacaacagat aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt atttgatgcc 3181 tggcagttcc ctactctcgc gttaacgcta gcatggatgt tttcccagtc acgacgttgt 3241 aaaacgacgg ccagtcttaa gctcgggccc //