LOCUS V018478 8041 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V018478 VERSION V018478 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 649..2721 /label="FBXL5(NM_012161)" /note="FBXL5(NM_012161)" /gene="FBXL5" CDS 2728..2793 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 2817..3405 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 3471..3519 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3721..4149 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4163..4492 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4559..5353 /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 5529..5650 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5699..5715) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5723..5739 /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(5747..5777) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5792..5813 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6101..6689) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6860..7720) /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(7721..7825) /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 gcgccaccat ggcgcccttt 661 cctgaagaag tggacgtctt caccgcccca cactggcgga tgaagcagct ggtggggctc 721 tactgcgaca agctttctaa aaccaatttt tccaacaaca acgatttccg tgctcttctg 781 cagtctttgt atgctacttt caaggagttc aaaatgcatg agcagattga aaatgaatac 841 attattggtt tgcttcaaca acgcagccag accatttata atgtacattc tgacaataaa 901 ctctccgaga tgcttagcct ctttgaaaag ggactgaaga atgttaagaa tgaatatgaa 961 cagttaaatt atgcaaaaca actgaaagag agattggagg cttttacaag agattttctt 1021 cctcacatga aagaggaaga ggaggttttt cagcccatgt taatggaata ttttacctat 1081 gaagagctta aggatattaa aaagaaagtg attgcacaac actgctctca gaaggatact 1141 gcagaactcc ttagaggtct tagcctatgg aatcatgctg aagagcgaca gaagtttttt 1201 aaatattccg tggatgaaaa gtcagataaa gaagcagaag tgtcagaaca ctccacaggt 1261 ataacccatc ttcctcctga ggtaatgctg tcaattttca gctatcttaa tcctcaagag 1321 ttatgtcgat gcagtcaagt aagcatgaaa tggtctcagc tgacaaaaac gggatcgctt 1381 tggaaacatc tttaccctgt tcattgggcc agaggtgact ggtatagtgg tcccgcaact 1441 gaacttgata ctgaacctga tgatgaatgg gtgaaaaata ggaaagatga aagtcgtgct 1501 tttcatgagt gggatgaaga tgctgacatt gatgaatctg aagagtctgc ggaggaatca 1561 attgctatca gcattgcaca aatggaaaaa cgtttactcc atggcttaat tcataacgtt 1621 ctaccatatg ttggtacttc tgtaaaaacc ttagtattag catacagctc tgcagtttcc 1681 agcaaaatgg ttaggcagat tttagagctt tgtcctaacc tggagcatct ggatcttacc 1741 cagactgaca tttcagattc tgcatttgac agttggtctt ggcttggttg ctgccagagt 1801 cttcggcatc ttgatctgtc tggttgtgag aaaatcacag atgtggccct agagaagatt 1861 tccagagctc ttggaattct gacatctcat caaagtggct ttttgaaaac atctacaagc 1921 aaaattactt caactgcgtg gaaaaataaa gacattacca tgcagtccac caagcagtat 1981 gcctgtttgc acgatttaac taacaagggc attggagaag aaatagataa tgaacacccc 2041 tggactaagc ctgtttcttc tgagaatttc acttctcctt atgtgtggat gttagatgct 2101 gaagatttgg ctgatattga agatactgtg gaatggagac atagaaatgt tgaaagtctt 2161 tgtgtaatgg aaacagcatc caactttagt tgttccacct ctggttgttt tagtaaggac 2221 attgttggac taaggactag tgtctgttgg cagcagcatt gtgcttctcc agcctttgcg 2281 tattgtggtc actcattttg ttgtacagga acagctttaa gaactatgtc atcactccca 2341 gaatcttctg caatgtgtag aaaagcagca aggactagat tgcctagggg aaaagactta 2401 atttactttg ggagtgaaaa atctgatcaa gagactggac gtgtacttct gtttctcagt 2461 ttatctggat gttatcagat cacagaccat ggtctcaggg ttttgactct gggaggaggg 2521 ctgccttatt tggagcacct taatctctct ggttgtctta ctataactgg tgcaggcctg 2581 caggatttgg tttcagcatg tccttctctg aatgatgaat acttttacta ctgtgacaac 2641 attaacggtc ctcatgctga taccgccagt ggatgccaga atttgcagtg tggttttcga 2701 gcctgctgcc gctctggcga actcgaggac tacaaagacc atgacggtga ttataaagat 2761 catgacatcg actacaagga tgacgatgac aagtagtgag ggcccgatat ctcgacaatc 2821 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt 2881 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg 2941 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc 3001 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt 3061 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg 3121 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg 3181 gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct 3241 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc 3301 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc 3361 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga aacgggggag 3421 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa 3481 aagacagaat aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg gttcggtccc 3541 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt 3601 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg 3661 tcggggcggc aggccctgcc atagcagatc tgcgcagctg gggctctagg gggtatcccc 3721 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg 3781 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca 3841 cgttcgccgg ctttccccgt caagctctaa atcggggcat ccctttaggg ttccgattta 3901 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc 3961 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg 4021 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat 4081 aagggatttt ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta 4141 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc 4201 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc 4261 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat 4321 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc 4381 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga 4441 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc 4501 gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat 4561 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg 4621 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc 4681 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca 4741 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct 4801 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga 4861 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg 4921 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat 4981 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga 5041 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg 5101 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg 5161 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat 5221 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct 5281 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga 5341 cgagttcttc tgagcgggac tctggggttc gcgaaatgac cgaccaagcg acgcccaacc 5401 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg 5461 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg 5521 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa 5581 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca 5641 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt 5701 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg 5761 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt 5821 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg 5881 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg 5941 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa 6001 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc 6061 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc 6121 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 6181 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc 6241 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct 6301 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 6361 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 6421 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga 6481 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa 6541 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta 6601 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc 6661 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 6721 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 6781 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 6841 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct 6901 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg 6961 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 7021 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa 7081 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 7141 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt 7201 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 7261 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt 7321 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 7381 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt 7441 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata 7501 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga 7561 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag 7621 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa 7681 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt 7741 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga 7801 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg 7861 gatcgggaga tctcccgatc ccctatggtc gactctcagt acaatctgct ctgatgccgc 7921 atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag 7981 caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag 8041 g //