LOCUS pEN-R2-GGGGS-miniTurbo-FLAG-L3 3454 bp ds-DNA circular SYN 20-MAR-2023 DEFINITION . ACCESSION VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM recombinant plasmid. REFERENCE 1 (bases 1 to 3454) AUTHORS VIB Network TITLE Direct Submission JOURNAL Exported Sep 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 linker" CDS 143..913 /codon_start=1 /translation="IPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSGDAC IAEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKLGA DKVRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGWI TLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGD KEIFGISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK" /label="miniTurbo" tag 914..937 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /translation="DYKDDDDK" /modified_by="mamee" /label="FLAG" attL3 complement(941..1036) /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(1094..1114) /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 1175..1225 /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 1226..2035 /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 2182..2770 /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 3369..3389 /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 cgatcccgct gctgaacgct aaacagattc tgggacagct 181 ggacggcggg agcgtggcag tcctgcctgt ggtcgactcc accaatcagt acctgctgga 241 tcgaatcggc gagctgaaga gtggggatgc ttgcattgca gaatatcagc aggcagggag 301 aggaagcaga gggaggaaat ggttctctcc ttttggagct aacctgtacc tgagtatgtt 361 ttggcgcctg aagcggggac cagcagcaat cggcctgggc ccggtcatcg gaattgtcat 421 ggcagaagcg ctgcgaaagc tgggagcaga caaggtgcga gtcaaatggc ccaatgacct 481 gtatctgcag gatagaaagc tggcaggcat cctggtggag ctggccggaa taacaggcga 541 tgctgcacag atcgtcattg gcgccgggat taacgtggct atgaggcgcg tggaggaaag 601 cgtggtcaat cagggctgga tcacactgca ggaagcaggg attaacctgg acaggaatac 661 tctggccgct atgctgatcc gagagctgcg ggcagccctg gaactgttcg agcaggaagg 721 cctggctcca tatctgtcac ggtgggagaa gctggataac ttcatcaata gacccgtgaa 781 gctgatcatt ggggacaaag agattttcgg gattagccgg gggattgata aacagggagc 841 cctgctgctg gaacaggacg gagttatcaa accctggatg ggcggagaaa tcagtctgcg 901 gtctgccgaa aaggactaca aagacgatga cgataaatga caactttatt atacaaagtt 961 ggcattataa aaaagcattg cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa 1021 aataaaatca ttatttggag ctccatggta gcgttaacgc ggccgcgata tcccctatag 1081 tgagtcgtat tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct 1141 ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta 1201 cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg 1261 attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg 1321 gcaatcaggt gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct 1381 gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg 1441 gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc 1501 atggttactc accactgcga tccccggaaa aacagcattc caggtattag aagaatatcc 1561 tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat 1621 tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc 1681 acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc 1741 tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt 1801 cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg 1861 tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa 1921 ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga 1981 taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga 2041 attggttaat tggttgtaac actggcagag cattacgctg acttgacggg acggcgcaag 2101 ctcatgacca aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt 2161 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca 2221 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct 2281 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta 2341 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct 2401 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc 2461 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca 2521 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agcattgaga 2581 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg 2641 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt 2701 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag 2761 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt 2821 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt 2881 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga 2941 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta 3001 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa 3061 tacgcgtacc gctagccagg aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg 3121 gccttctgct tagtttgatg cctggcagtt tatggcgggc gtcctgcccg ccaccctccg 3181 ggccgttgct tcacaacgtt caaatccgct cccggcggat ttgtcctact caggagagcg 3241 ttcaccgaca aacaacagat aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt 3301 atttgatgcc tggcagttcc ctactctcgc gttaacgcta gcatggatgt tttcccagtc 3361 acgacgttgt aaaacgacgg ccagtcttaa gctcgggccc tgcagctcta gagctcgaat 3421 tctacaggtc actaatacca tctaagtagt tgat //