LOCUS pEN-R2-GGGGS-Turbo-FLAG-L3 3640 bp ds-DNA circular SYN 20-MAR-2023 DEFINITION . ACCESSION VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM recombinant plasmid. REFERENCE 1 (bases 1 to 3640) AUTHORS VIB Network TITLE Direct Submission JOURNAL Exported Jul 10, 2020 from SnapGene 4.2.0 http://www.snapgene.com FEATURES Location/Qualifiers attR2 complement(1..126) /created_by="mamee" /Color="#00cccc" /modified_by="mamee" /note="Derived using Geneious Prime 2023.0.1 'Annotate from Database' based on nucleotide similarity" /Transferred_From="attR2" /Transferred_Similarity="99.21%" /Primary_Match="99.21%; Adjusted-Interval-0:0/0; attR2" CDS 128..142 /codon_start=1 /translation="GGGGS" /label="GGGGS" CDS 143..1099 /created_by="mamee" /label="Turbo" tag 1100..1123 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /translation="DYKDDDDK" /modified_by="mamee" /label="FLAG" attL3 complement(1127..1222) /created_by="mamee" /Color="#00cccc" /modified_by="mamee" /note="Derived using Geneious Prime 2023.0.1 '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 complement(1280..1300) /note="Derived using Geneious Prime 2020.2.2 '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 1361..1411 /created_by="chgai" /note="Derived using Geneious Prime 2022.1.1 'Annotate from Database' based on nucleotide similarity" /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 2023.0.1 '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 1412..2221 /modified_by="wacae" /note="Derived using Geneious Prime 2020.2.2 '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" rep_origin 2368..2956 /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="99.49%" /Primary_Match="99.49%; 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 3555..3575 /note="Derived using Geneious Prime 2020.2.2 '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 tcatagtgac tgcatatgtt gtgttttaca gtattatgta gtctgttttt tatgcaaaat 61 ctaatttaat atattgatat ttatatcatt ttacgtttct cgttcaactt tcttgtacaa 121 agtggccgga ggcggtggat cgaaagacaa tactgtgcct ctgaagctga tcgctctcct 181 ggctaatggc gagttccata gtggcgaaca gctgggagaa accctgggca tgtccagggc 241 cgctatcaac aagcacattc agactctgcg cgactggggc gtggacgtgt tcaccgtgcc 301 cggaaagggc tactctctgc ccgagcctat cccgctgctg aacgctaaac agattctggg 361 acagctggac ggcgggagcg tggcagtcct gcctgtggtc gactccacca atcagtacct 421 gctggatcga atcggcgagc tgaagagtgg ggatgcttgc attgcagaat atcagcaggc 481 agggagagga agcagaggga ggaaatggtt ctctcctttt ggagctaacc tgtacctgag 541 tatgttttgg cgcctgaagc ggggaccagc agcaatcggc ctgggcccgg tcatcggaat 601 tgtcatggca gaagcgctgc gaaagctggg agcagacaag gtgcgagtca aatggcccaa 661 tgacctgtat ctgcaggata gaaagctggc aggcatcctg gtggagctgg ccggaataac 721 aggcgatgct gcacagatcg tcattggcgc cgggattaac gtggctatga ggcgcgtgga 781 ggaaagcgtg gtcaatcagg gctggatcac actgcaggaa gcagggatta acctggacag 841 gaatactctg gccgctacgc tgatccgaga gctgcgggca gccctggaac tgttcgagca 901 ggaaggcctg gctccatatc tgccacggtg ggagaagctg gataacttca tcaatagacc 961 cgtgaagctg atcattgggg acaaagagat tttcgggatt agccggggga ttgataaaca 1021 gggagccctg ctgctggaac aggacggagt tatcaaaccc tggatgggcg gagaaatcag 1081 tctgcggtct gccgaaaagg actacaaaga cgatgacgat aaatgacaac tttattatac 1141 aaagttggca ttataaaaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc 1201 agtcaaaata aaatcattat ttggagctcc atggtagcgt taacgcggcc gcgatatccc 1261 ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca 1321 aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc 1381 tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag 1441 gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa 1501 tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt 1561 gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact 1621 aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga 1681 tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga 1741 atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca 1801 ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc 1861 gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg 1921 ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc 1981 agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat 2041 aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct 2101 atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg 2161 tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta 2221 atcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg 2281 cgcaagctca tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga 2341 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg 2401 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc 2461 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct 2521 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc 2581 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt 2641 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg 2701 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagca 2761 ttgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag 2821 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag 2881 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg 2941 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg 3001 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac 3061 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt 3121 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat 3181 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc 3241 aattaatacg cgtaccgcta gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc 3301 aggatggcct tctgcttagt ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac 3361 cctccgggcc gttgcttcac aacgttcaaa tccgctcccg gcggatttgt cctactcagg 3421 agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtcttccga ctgagccttt 3481 cgttttattt gatgcctggc agttccctac tctcgcgtta acgctagcat ggatgttttc 3541 ccagtcacga cgttgtaaaa cgacggccag tcttaagctc gggccctgca gctctagagc 3601 tcgaattcta caggtcacta ataccatcta agtagttgat //