LOCUS Exported 4668 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Localization: C1 Cloning Vector, Excitation: 551, Emission: 565. ACCESSION . VERSION . KEYWORDS mKO2-C1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4668) AUTHORS Sakaue-Sawano A, Kurokawa H, Morimura T, Hanyu A, Hama H, Osawa H, Kashiwagi S, Fukami K, Miyata T, Miyoshi H, Imamura T, Ogawa M, Masai H, Miyawaki A TITLE Visualizing spatiotemporal dynamics of multicellular cell-cycle progression. JOURNAL Cell. 2008 Feb 8;132(3):487-98. doi: 10.1016/j.cell.2007.12.033. PUBMED 18267078 REFERENCE 2 (bases 1 to 4668) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4668 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 61..364 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 365..568 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 519..539 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 607..616 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 613..1266 /codon_start=1 /product="monomeric Kusabira-Orange 2 fluorescent protein" /label=mKO2 /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY LKLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS" misc_feature 1267..1332 /label=MCS /note="multiple cloning site" polyA_signal 1456..1577 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1493..1512) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1547..1566 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(1584..2039) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1721..1742) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 1933..1952 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2066..2170 /gene="bla" /label=AmpR promoter promoter 2172..2529 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2195..2215) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2380..2515 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2442..2461 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2564..3358 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(2618..2637) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3228..3247 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(3546..3565) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 3590..3637 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 3966..4554 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4455..4474 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg 61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta 601 ccggtcgcca ccatggtgag tgtgattaaa ccagagatga agatgaggta ctacatggac 661 ggctccgtca atgggcatga gttcacaatt gaaggtgaag gcacaggcag accttacgag 721 ggacatcaag agatgacact acgcgtcaca atggccgagg gcgggccaat gcctttcgcg 781 tttgacttag tgtcacacgt gttctgttac ggccacagag tatttactaa atatccagaa 841 gagataccag actatttcaa acaagcattt cctgaaggcc tgtcatggga aaggtcgttg 901 gagttcgaag atggtgggtc cgcttcagtc agtgcgcata taagccttag aggaaacacc 961 ttctaccaca aatccaaatt tactggggtt aactttcctg ccgatggtcc tatcatgcaa 1021 aaccaaagtg ttgattggga gccatcaacc gagaaaatta ctgccagcga cggagttctg 1081 aagggtgatg ttacgatgta cctaaaactt gaaggaggcg gcaatcacaa atgccaaatg 1141 aagactactt acaaggcggc aaaagagatt cttgaaatgc caggagacca ttacatcggc 1201 catcgcctcg tcaggaaaac cgaaggcaac attactgagc aggtagaaga tgcagtagct 1261 catagctccg gactcagatc tcgagctcaa gcttcgaatt ctgcagtcga cggtaccgcg 1321 ggcccgggat ccaccggatc tagataactg atcataatca gccataccac atttgtagag 1381 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat 1441 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc 1501 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa 1561 ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt 1621 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta 1681 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc 1741 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg 1801 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact 1861 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt 1921 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc 1981 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc 2041 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca 2101 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa 2161 aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga 2221 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca 2281 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc 2341 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc 2401 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag 2461 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc 2521 ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg 2581 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca 2641 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt 2701 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg 2761 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga 2821 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca 2881 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct 2941 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac 3001 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc 3061 gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt 3121 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt 3181 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg 3241 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat 3301 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc 3361 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc 3421 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc 3481 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg 3541 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa 3601 agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag 3661 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct 3721 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc 3781 ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa 3841 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa 3901 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga 3961 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg 4021 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact 4081 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac 4141 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg 4201 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg 4261 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga 4321 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc 4381 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg 4441 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc 4501 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc 4561 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt 4621 cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcat //