LOCUS V019454 7667 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V019454 VERSION V019454 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 671..2347 /label="YTHDF1(NM_017798)" /note="YTHDF1(NM_017798)" /gene="YTHDF1" CDS 2354..2419 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 2443..3031 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 3097..3145 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3347..3775 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3789..4118 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4185..4979 /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 5155..5276 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5325..5341) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5349..5365 /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(5373..5403) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5418..5439 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5727..6315) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6486..7346) /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(7347..7451) /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 taccgccacc atgtcggcca ccagcgtgga cacccagaga acaaaaggac aagataataa 721 agtacaaaat ggttcgttac atcagaagga tacagttcat gacaatgact ttgagcccta 781 ccttactgga cagtcaaatc agagtaacag ttacccctca atgagcgacc cctacctgtc 841 cagctattac ccgccgtcca ttggatttcc ttactccctc aatgaggctc cgtggtctac 901 tgcaggggac cctccgattc catacctcac cacctacgga cagctcagta acggagacca 961 tcattttatg cacgatgctg tttttgggca gcctgggggc ctggggaaca acatctatca 1021 gcacaggttc aattttttcc ctgaaaaccc tgcgttctca gcatggggga caagtgggtc 1081 tcaaggtcag cagacccaga gctccgcgta tgggagcagc tacacctacc ccccgagctc 1141 cctgggtggc acggtggttg atgggcagcc aggctttcac agcgacaccc tcagcaaggc 1201 ccccgggatg aacagcctgg agcagggcat ggttggcctg aagattgggg acgtcagctc 1261 ctccgccgtc aagacggtgg gctctgtcgt cagcagcgtg gcactgactg gtgtcctttc 1321 tggcaacggt gggacaaatg tgaacatgcc agtttcaaag ccgacctcgt gggctgccat 1381 tgccagcaag cctgcaaaac cacagcctaa aatgaaaaca aagagcgggc ctgtcatggg 1441 gggtgggctg ccccctccac ccataaagca taacatggac attggcacct gggataacaa 1501 ggggcctgtg ccgaaggccc cagtccccca gcaggcaccc tctccacagg ctgccccaca 1561 gccccagcag gtggctcagc ctctcccagc acagccccca gctttggctc aaccgcagta 1621 tcagagccct cagcagccac cccagacccg ctgggttgcc ccacgcaaca gaaacgcggc 1681 gtttgggcag agcggagggg ctggcagcga tagcaactct cctggaaacg tccagcctaa 1741 ttctgccccc agcgtcgaat cccaccccgt ccttgaaaaa ctgaaggctg ctcacagcta 1801 caacccgaaa gagtttgagt ggaatctgaa aagcgggcgt gtgttcatca tcaagagcta 1861 ctctgaggac gacatccacc gctccattaa gtactccatc tggtgtagca cagagcacgg 1921 caacaagcgc ctggacagcg ccttccgctg catgagcagc aaggggcccg tctacctgct 1981 cttcagcgtc aatgggagtg ggcatttttg tggggtggcc gagatgaagt cccccgtgga 2041 ctacggcacc agtgccgggg tctggtctca ggacaagtgg aaggggaagt ttgatgtcca 2101 gtggattttt gttaaggatg tacccaataa ccagctccgg cacatcaggc tggagaataa 2161 cgacaacaaa ccggtcacaa actcccggga cacccaggag gtgcccttag aaaaagccaa 2221 gcaagtgctg aaaattatca gttcctacaa gcacacaacc tccatcttcg acgactttgc 2281 tcactacgag aagcgccagg aggaggagga ggtggtgcgc aaggaacggc agagtcgaaa 2341 caaacaactc gaggactaca aagaccatga cggtgattat aaagatcatg acatcgacta 2401 caaggatgac gatgacaagt agtgagggcc cgatatctcg acaatcaacc tctggattac 2461 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga 2521 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc 2581 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa 2641 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc 2701 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc 2761 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc 2821 gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg 2881 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct 2941 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg 3001 agtcggatct ccctttgggc cgcctccccg cctggaaacg ggggaggcta actgaaacac 3061 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa 3121 cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct 3181 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca 3241 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc 3301 cctgccatag cagatctgcg cagctggggc tctagggggt atccccacgc gccctgtagc 3361 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc 3421 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt 3481 ccccgtcaag ctctaaatcg gggcatccct ttagggttcc gatttagtgc tttacggcac 3541 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag 3601 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa 3661 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgggg 3721 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc 3781 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta 3841 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag 3901 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa 3961 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac 4021 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt 4081 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat 4141 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg 4201 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac 4261 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg 4321 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc 4381 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg 4441 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc 4501 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc 4561 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta 4621 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg 4681 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg 4741 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat 4801 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc 4861 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta 4921 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag 4981 cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat 5041 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc 5101 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg 5161 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa 5221 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat 5281 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct 5341 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt 5401 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc 5461 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg 5521 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg 5581 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca 5641 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac 5701 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac 5761 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg 5821 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac 5881 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat 5941 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag 6001 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac 6061 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt 6121 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt 6181 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc 6241 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga 6301 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac 6361 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc 6421 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct 6481 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca 6541 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct 6601 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca 6661 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc 6721 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg 6781 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct 6841 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa 6901 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta 6961 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc 7021 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg 7081 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa 7141 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg 7201 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc 7261 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg 7321 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat 7381 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata 7441 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc 7501 ccgatcccct atggtcgact ctcagtacaa tctgctctga tgccgcatag ttaagccagt 7561 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta 7621 caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagg //