LOCUS V020777 7081 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V020777 VERSION V020777 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 700..1734 /label="FOXL1(NM_005250)" /note="FOXL1(NM_005250)" /gene="FOXL1" CDS 1768..1833 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 1857..2445 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2511..2559 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2761..3189 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3203..3532 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3599..4393 /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 4569..4690 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4739..4755) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4763..4779 /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(4787..4817) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4832..4853 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5141..5729) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5900..6760) /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(6761..6865) /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 ggatccacta gtccagtgtg gtggccacca tgagtcacct cttcgatccc 721 cggctgcctg ccctggccgc ctcgcccatg ctgtatctgt acggtcccga gagacccggc 781 ctccctctgg ccttcgcccc cgcggctgct ctagctgcct cgggccgggc cgagaccccg 841 cagaagcctc cctacagcta catcgcgctc atcgccatgg cgatccagga cgcgcccgag 901 cagagggtca cgctcaacgg catctaccag ttcatcatgg accgcttccc cttctaccac 961 gacaaccggc agggctggca gaacagcatc cgccacaacc tctcgctcaa cgactgcttc 1021 gtcaaggtgc cccgcgagaa agggcggccg ggcaagggca gctactggac gctggacccc 1081 cgctgcctgg acatgtttga gaacggcaac taccggcgcc ggaagaggaa gcccaagccg 1141 ggccccgggg ccccggaggc caagaggccc cgcgccgaga cgcaccagcg cagcgcggag 1201 gcgcagccgg aggcggggag cggggcaggg ggctcgggcc ccgcaatctc ccgcctgcag 1261 gcagcgcccg cgggcccctc gcccctcctg gacggcccct ctccgccggc gcccctccac 1321 tggccgggga ccgcgtcccc gaacgaggac gctggtgacg ctgcccaggg cgcagcggcc 1381 gtggcggtcg gccaggcagc gcgcacaggg gacggcccgg ggtcccctct gcgccccgcc 1441 tcccgcagct ctccgaagag ctccgacaag tccaagagct tcagcataga cagcatcctg 1501 gcgggaaagc agggccagaa gccgccttca ggggacgaac tcctaggggg tgccaagcct 1561 gggcccggcg gccgtctggg tgcctcgctc ctggccgcct cctccagcct ccgtccgcct 1621 ttcaacgctt ccctgatgct cgacccgcat gtccagggcg gcttttacca gctcgggatc 1681 cccttcctct cttatttccc cctgcaggtt cccgacacgg tactccactt ccagtgcaga 1741 tatccagcac agtggcggcc gctcgaggac tacaaagacc atgacggtga ttataaagat 1801 catgacatcg actacaagga tgacgatgac aagtagtgag ggcccgatat ctcgacaatc 1861 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt 1921 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg 1981 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc 2041 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt 2101 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg 2161 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg 2221 gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct 2281 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc 2341 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc 2401 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga aacgggggag 2461 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa 2521 aagacagaat aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg gttcggtccc 2581 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt 2641 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg 2701 tcggggcggc aggccctgcc atagcagatc tgcgcagctg gggctctagg gggtatcccc 2761 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg 2821 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca 2881 cgttcgccgg ctttccccgt caagctctaa atcggggcat ccctttaggg ttccgattta 2941 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc 3001 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg 3061 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat 3121 aagggatttt ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta 3181 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc 3241 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc 3301 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat 3361 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc 3421 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga 3481 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc 3541 gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat 3601 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg 3661 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc 3721 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca 3781 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct 3841 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga 3901 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg 3961 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat 4021 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga 4081 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg 4141 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg 4201 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat 4261 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct 4321 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga 4381 cgagttcttc tgagcgggac tctggggttc gcgaaatgac cgaccaagcg acgcccaacc 4441 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg 4501 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg 4561 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa 4621 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca 4681 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt 4741 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg 4801 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt 4861 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg 4921 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg 4981 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa 5041 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc 5101 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc 5161 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 5221 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc 5281 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct 5341 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 5401 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 5461 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga 5521 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa 5581 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta 5641 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc 5701 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 5761 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 5821 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 5881 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct 5941 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg 6001 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 6061 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa 6121 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 6181 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt 6241 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 6301 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt 6361 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 6421 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt 6481 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata 6541 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga 6601 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag 6661 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa 6721 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt 6781 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga 6841 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg 6901 gatcgggaga tctcccgatc ccctatggtc gactctcagt acaatctgct ctgatgccgc 6961 atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag 7021 caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag 7081 g //