LOCUS V019501 7757 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V019501 VERSION V019501 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..2437 /label="YTHDF3(NM_152758)" /note="YTHDF3(NM_152758)" /gene="YTHDF3" CDS 2444..2509 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 2533..3121 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 3187..3235 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3437..3865 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3879..4208 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4275..5069 /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 5245..5366 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5415..5431) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5439..5455 /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(5463..5493) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5508..5529 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5817..6405) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6576..7436) /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(7437..7541) /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 ccatgtcagc cactagcgtg gatcagagac ctaaagggca 721 aggaaataaa gtttcagtac aaaacggttc gattcatcaa aaagatgctg taaatgatga 781 tgattttgag ccatacttaa gtagccagac aaatcagagt aacagctatc caccaatgtc 841 agatccatac atgcctagtt actatgctcc atccattgga tttccatatt ctcttgggga 901 agcagcgtgg tccacagctg gagaccagcc tatgccatat ctgacaacct atggacaaat 961 gagtaatgga gaacatcact atataccaga tggtgtattt agtcaacctg gggcattagg 1021 aaatacccct ccatttcttg gtcaacatgg atttaacttt tttcctggta atgctgattt 1081 ctctacatgg gggacaagtg gatctcaggg acaatcaaca caaagttctg cttatagtag 1141 cagttatggc tatccaccta gttctcttgg gagagctatt actgatggac aggctggatt 1201 tggcaatgat actttgagta aggtgcctgg cattagcagt attgagcaag gcatgactgg 1261 actgaaaatt ggtggtgacc tgacagctgc agtgacaaaa actgtaggta cagctttgag 1321 cagcagtggt atgactagca ttgcaaccaa tagtgtgccc ccagttagca gtgcagcacc 1381 taaaccaacc tcctgggctg ccattgccag aaagcctgcc aaacctcaac cgaaacttaa 1441 acccaagggc aatgtgggaa ttgggggttc tgctgtacca ccacctccta taaaacacaa 1501 catgaatatt ggaacttggg atgaaaaagg gtcagtggta aaggctccac caacccaacc 1561 agttctgcct cctcaaacta taatccagca gcctcagcca ttaattcaac caccaccatt 1621 ggtgcaaagc caactgcctc aacagcagcc tcaaccacca caaccacagc agcaacaagg 1681 acctcagcca caggcccagc ctcaccaagt gcagcctcaa cagcagcagc tgcagaatcg 1741 ctgggtagct cctcgtaaca ggggagcagg cttcaaccag aacaatggag cgggcagtga 1801 aaactttggt ttaggtgttg tacctgtcag tgcttcacct tctagtgtag aagtgcatcc 1861 cgtgctggaa aagctaaagg ccataaacaa ctataatccc aaagactttg attggaatct 1921 gaagaatgga cgtgtgttta taattaaaag ctactctgag gatgacatac atcgttccat 1981 taaatactct atctggtgta gtactgagca tggtaataag cgtttggatg cagcttaccg 2041 ttccctgaat gggaaaggcc cactctattt actcttcagt gtgaatggca gtggacattt 2101 ttgtggagtg gctgaaatga agtctgttgt ggactataat gcgtatgctg gtgtctggtc 2161 tcaggataag tggaagggca aatttgaagt taaatggatc tttgtcaaag atgttcccaa 2221 taaccaatta cggcatattc gcttagaaaa taatgacaac aaaccggtta ccaattcaag 2281 ggacactcaa gaggtacccc tagaaaaagc taagcaagtg cttaaaataa ttgctacttt 2341 caagcatacc acctcaatct ttgatgactt tgcacattat gaaaagcgtc aagaagagga 2401 ggaagccatg cgtagggaga gaaatagaaa caaacaactc gaggactaca aagaccatga 2461 cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt agtgagggcc 2521 cgatatctcg acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt 2581 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct 2641 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt 2701 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac 2761 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct 2821 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca 2881 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt 2941 ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc 3001 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct 3061 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg 3121 cctggaaacg ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg 3181 ctatgacggc aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa 3241 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc 3301 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca 3361 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cagatctgcg cagctggggc 3421 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt 3481 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc 3541 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg gggcatccct 3601 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat 3661 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc 3721 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc 3781 tattcttttg atttataagg gattttgggg atttcggcct attggttaaa aaatgagctg 3841 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa 3901 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa 3961 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca 4021 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca 4081 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg 4141 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct 4201 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat 4261 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg 4321 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg 4381 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg 4441 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc 4501 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg 4561 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca 4621 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc 4681 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg 4741 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg 4801 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata 4861 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg 4921 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat 4981 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct 5041 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgcga aatgaccgac 5101 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg 5161 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc 5221 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa 5281 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt 5341 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc 5401 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca 5461 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa 5521 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag 5581 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc 5641 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct 5701 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg 5761 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc 5821 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga 5881 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct 5941 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg 6001 gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag 6061 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat 6121 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac 6181 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac 6241 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc 6301 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt 6361 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc 6421 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg 6481 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca 6541 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca 6601 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag 6661 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac 6721 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc 6781 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct 6841 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc 6901 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg 6961 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc 7021 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat 7081 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag 7141 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat 7201 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg 7261 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca 7321 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga 7381 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc 7441 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata 7501 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg 7561 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtcgact ctcagtacaa 7621 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg 7681 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca 7741 tgaagaatct gcttagg //