LOCUS V024787 7711 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V024787 VERSION V024787 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 775..2367 /label="CYP27A1(NM_000784)" /note="CYP27A1(NM_000784)" /gene="CYP27A1" CDS 2368..2385 /codon_start=1 /product="6xHis affinity tag" /transl_table=1 /translation="HHHHHH" /label="6xHis" misc_feature 2487..3075 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 3141..3189 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3391..3819 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3833..4162 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4229..5023 /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 5199..5320 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5369..5385) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5393..5409 /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(5417..5447) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5462..5483 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5771..6359) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6530..7390) /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(7391..7495) /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 gctcgtttag tgaaccgtca gatcgcctgg 661 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg 721 actctagagc tagcgtttaa acttaagctt ggtaccgagc tcggatccgc caccatggct 781 gcgctgggct gcgcgaggct gaggtgggcg ctgcgagggg ccggccgtgg cctctgcccc 841 cacggggcca gagccaaggc cgcgatccct gccgccctcc cctcggacaa ggccaccgga 901 gctcccggag ccgggcctgg tgtccggcgg cggcaacgga gcttagagga gattccacgt 961 ctaggacagc tgcgcttctt ctttcagctg ttcgttcaag gctatgccct gcaactgcac 1021 cagttacagg tgctttacaa ggccaagtac ggtccaatgt ggatgtccta cttagggcct 1081 cagatgcacg tgaacctggc cagtgccccg ctcttggagc aagtgatgcg gcaagagggc 1141 aagtacccag tacggaacga catggagcta tggaaggagc accgggacca gcacgacctg 1201 acctatgggc cgttcaccac ggaaggacac cactggtacc agctgcgcca ggctctgaac 1261 cagcggttgc tgaagccagc ggaagcagcg ctctatacgg atgctttcaa tgaggtgatt 1321 gatgacttta tgactcgact ggaccagctg cgggcagaga gtgcttcggg gaaccaggtg 1381 tcggacatgg ctcaactctt ctactacttt gccttggaag ctatttgcta catcctgttc 1441 gagaaacgca ttggctgcct gcagcgatcc atccccgagg acaccgtgac cttcgtcaga 1501 tccatcgggt taatgttcca gaactcactc tatgccacct tcctccccaa gtggactcgc 1561 cccgtgctgc ctttctggaa gcgatacctg gatggttgga atgccatctt ttcctttggg 1621 aagaagctga ttgatgagaa gctcgaagat atggaggccc aactgcaggc agcagggcca 1681 gatggcatcc aggtgtctgg ctacctgcac ttcttactgg ccagtggaca gctcagtcct 1741 cgggaggcca tgggcagcct gcctgagctg ctcatggctg gagtggacac gacatccaac 1801 acgctgacat gggccctgta ccacctctca aaggaccctg agatccagga ggccttgcac 1861 gaggaagtgg tgggtgtggt gccagccggg caagtgcccc agcacaagga ctttgcccac 1921 atgccgttgc tcaaagctgt gcttaaggag actctgcgtc tctaccctgt ggtccccaca 1981 aactcccgga tcatagaaaa ggaaattgaa gttgatggct tcctcttccc caagaacacc 2041 cagtttgtgt tctgccacta tgtggtgtcc cgggacccca ctgccttctc tgagcctgaa 2101 agcttccagc cccaccgctg gctgagaaac agccagcctg ctacccccag gatccagcac 2161 ccatttggct ctgtgccctt tggctatggg gtccgggcct gcctgggccg caggattgca 2221 gagctggaga tgcagctact cctcgcaagg ctgatccaga agtacaaggt ggtcctggcc 2281 ccggagacgg gggagttgaa gagtgtggcc cgcattgtcc tggttcccaa taagaaagtg 2341 ggcctgcagt tcctgcagag acagtgccac catcaccatc accattagaa ttctgcagat 2401 atccagcaca gtggcggccg ctcgagtcta gagggcccct taagggttcg atccctaccg 2461 gttagtaatg agtttgatat ctcgacaatc aacctctgga ttacaaaatt tgtgaaagat 2521 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc 2581 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct 2641 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca 2701 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt 2761 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg 2821 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga 2881 agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt 2941 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc 3001 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt 3061 gggccgcctc cccgcctgga aacgggggag gctaactgaa acacggaagg agacaatacc 3121 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg ggtgttgggt 3181 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga 3241 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt 3301 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcagatc 3361 tgcgcagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg 3421 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc 3481 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa 3541 atcggggcat ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac 3601 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt 3661 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca 3721 accctatctc ggtctattct tttgatttat aagggatttt ggggatttcg gcctattggt 3781 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca 3841 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct 3901 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca 3961 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc 4021 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta 4081 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt 4141 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga 4201 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc 4261 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg 4321 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac 4381 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc 4441 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg 4501 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga 4561 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg 4621 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg 4681 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt 4741 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc 4801 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg 4861 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga 4921 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc 4981 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc 5041 gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg 5101 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc 5161 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata 5221 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc 5281 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga 5341 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc 5401 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct 5461 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa 5521 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta 5581 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc 5641 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg 5701 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt 5761 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa 5821 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct 5881 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc 5941 cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg 6001 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct 6061 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag 6121 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga 6181 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga 6241 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg 6301 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag 6361 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag 6421 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat 6481 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct 6541 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac 6601 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa 6661 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg 6721 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt 6781 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca 6841 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt 6901 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct 6961 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg 7021 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg 7081 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg 7141 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa 7201 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt 7261 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt 7321 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt 7381 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca 7441 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat 7501 ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtc 7561 gactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg 7621 tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt 7681 gaccgacaat tgcatgaaga atctgcttag g //