LOCUS V018851 6848 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V018851 VERSION V018851 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 50..429 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 430..633 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 683..1528 /label="ATF5(NM_012068)" /note="ATF5(NM_012068)" /gene="ATF5" CDS 1535..1600 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 1624..2212 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2278..2326 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2528..2956 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2970..3299 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3366..4160 /codon_start=1 /gene="aph(3')-II (or nptII)" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin®)" /product="aminoglycoside phosphotransferase from Tn5" /transl_table=1 /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" polyA_signal 4336..4457 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4506..4522) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4530..4546 /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG)." /label="lac operator" promoter complement(4554..4584) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4599..4620 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4908..5496) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5667..6527) /codon_start=1 /gene="bla" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /product="β-lactamase" /transl_table=1 /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA SLIKHW*" /label="AmpR" promoter complement(6528..6632) /gene="bla" /label="AmpR promoter" ORIGIN 1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta 61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag 121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc 181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga 241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat 301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc 361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct 421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca 481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat 541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg 601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg 661 taccgagctc ggatccgcca ccatgtcact cctggcgacc ctggggctgg agctggacag 721 ggccctgctc ccagctagtg ggctgggatg gctcgtagac tatgggaaac tccccccggc 781 ccctgccccc ctggctccct atgaggtcct tgggggagcc ctggagggcg ggcttccagt 841 ggggggagag cccctggcag gtgatggctt ctctgactgg atgactgagc gagttgattt 901 cacagctctc ctccctctgg agcctccctt accccccggc accctccccc aaccttcccc 961 aaccccacct gacctggaag ctatggcctc cctcctcaag aaggagctgg aacagatgga 1021 agacttcttc ctagatgccc cgcccctccc accaccctcc ccgccgccac taccaccacc 1081 accactacca ccagccccct ccctccccct gtccctcccc tcctttgacc tcccccagcc 1141 ccctgtcttg gatactctgg acttgctggc catctactgc cgcaacgagg ccgggcagga 1201 ggaagtgggg atgccgcctc tgcccccgcc acagcagccc cctcctcctt ctccacctca 1261 accttctcgc ctggccccct acccacatcc tgccaccacc cgaggggacc gcaagcaaaa 1321 gaagagagac cagaacaagt cggcggctct gaggtaccgc cagcggaagc gggcagaggg 1381 tgaggccctg gagggcgagt gccaggggct ggaggcacgg aatcgcgagc tgaaggaacg 1441 ggcagagtcc gtggagcgcg agatccagta cgtcaaggac ctgctcatcg aggtttacaa 1501 ggcccggagc cagaggaccc gtagctgcct cgaggactac aaagaccatg acggtgatta 1561 taaagatcat gacatcgact acaaggatga cgatgacaag tagtgagggc ccgatatctc 1621 gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt 1681 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc 1741 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag 1801 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc 1861 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc 1921 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg 1981 ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg 2041 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc 2101 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt 2161 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaaac 2221 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg 2281 caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa acgcggggtt 2341 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc 2401 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca 2461 gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg ctctaggggg 2521 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc 2581 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt 2641 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc tttagggttc 2701 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt 2761 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt 2821 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt 2881 gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct gatttaacaa 2941 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag 3001 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg 3061 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag 3121 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc 3181 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg 3241 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa 3301 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt 3361 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc 3421 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc 3481 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg 3541 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag 3601 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg 3661 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg 3721 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac 3781 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg 3841 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc 3901 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg 3961 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc 4021 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc 4081 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc 4141 ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga ccaagcgacg 4201 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc 4261 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag 4321 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc 4381 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa 4441 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa 4501 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata 4561 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta 4621 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa 4681 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg 4741 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag 4801 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa 4861 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc 4921 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca 4981 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg 5041 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct 5101 caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt 5161 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag 5221 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc 5281 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac 5341 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga 5401 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc 5461 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg 5521 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca 5581 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt 5641 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca 5701 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg 5761 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca 5821 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt 5881 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt 5941 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca 6001 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca 6061 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga 6121 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact 6181 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga 6241 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg 6301 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc 6361 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga 6421 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat 6481 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt 6541 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt 6601 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac 6661 gtcgacggat cgggagatct cccgatcccc tatggtcgac tctcagtaca atctgctctg 6721 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt 6781 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc 6841 tgcttagg //