LOCUS Exported 5167 bp ds-DNA circular SYN 18-OCT-2019 DEFINITION synthetic circular DNA. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5167) AUTHORS . TITLE Direct Submission JOURNAL Exported Monday, Oct 21, 2019 from SnapGene Viewer 5.0.4 https://www.snapgene.com FEATURES Location/Qualifiers source 1..5167 /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" CDS 687..1025 /codon_start=1 /label=HS22 /translation="MAQVQLQESGGGLAQAGGSLRLSCAASGIMFSAIETRWYRQVPGK QREWVADITSGGVTNYRPSVKGRFTISRDNAKNTIYLQMNSLKPEDTAIYYCASLPYWG QGTQVTVSS" CDS 1041..1058 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 1065..1091 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" regulatory 1107..1116 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1113..1832 /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" polyA_signal 1955..2076 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" rep_origin complement(2083..2538) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 2565..2669 /gene="bla" /label=AmpR promoter promoter 2671..3028 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2879..3014 /label=SV40 ori /note="SV40 origin of replication" CDS 3063..3857 /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" polyA_signal 4089..4136 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4465..5053 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" 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 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc gcgggccctg 661 gtttcgctac cgtggcccag gcggccatgg cccaggtgca gctgcaggag tcggggggag 721 gcttggcaca agctgggggg tctctgagac tctcctgcgc agcctctgga atcatgttca 781 gtgccattga aacgcgctgg taccgccagg ttccagggaa gcagcgcgag tgggtcgcag 841 atattactag tggtggggtc acaaactata gaccgtccgt gaagggccgg ttcaccatct 901 ccagagacaa cgccaagaac acgatatatc tgcaaatgaa cagcctgaaa cctgaggaca 961 cagccattta ttattgtgcg tcgctgcctt attggggcca ggggacccag gtcaccgtct 1021 cctcaggcca ggccggccag caccatcacc atcaccatgg cgcatacccg tacgacgttc 1081 cggactacgc tagggatcca ccggtcgcca ccatggtgag caagggcgag gagctgttca 1141 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg 1201 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca 1261 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccttcggc tacggcctgc 1321 agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc 1381 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc 1441 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg 1501 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca 1561 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc 1621 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg 1681 gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc gccctgagca 1741 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga 1801 tcactctcgg catggacgag ctgtacaagt aaagcggccg cgactctaga tcataatcag 1861 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa 1921 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg 1981 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc 2041 tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata 2101 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg 2161 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc 2221 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa 2281 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt 2341 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac 2401 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta 2461 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg 2521 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt 2581 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa 2641 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt 2701 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat 2761 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag 2821 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat 2881 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt 2941 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg 3001 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc 3061 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat 3121 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt 3181 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac 3241 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg 3301 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc 3361 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa 3421 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc 3481 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg 3541 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg 3601 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa 3661 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg 3721 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct 3781 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc 3841 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa 3901 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat 3961 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt 4021 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg 4081 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa 4141 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg 4201 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc 4261 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat 4321 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt 4381 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc 4441 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt 4501 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac 4561 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt 4621 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct 4681 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga 4741 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac 4801 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg 4861 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt 4921 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc 4981 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg 5041 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc 5101 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc 5161 catgcat //