LOCUS V018809 7112 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V018809 VERSION V018809 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..1759 /label="NANS(NM_018946)" /note="NANS(NM_018946)" /gene="NANS" CDS 1799..1864 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 1888..2476 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2542..2590 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2792..3220 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3234..3563 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3630..4424 /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 4600..4721 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4770..4786) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4794..4810 /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(4818..4848) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4863..4884 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5172..5760) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5931..6791) /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(6792..6896) /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 ccatgccgct ggagctggag ctgtgtcccg ggcgctgggt 721 gggcgggcaa cacccgtgct tcatcattgc cgagatcggc cagaaccacc agggcgacct 781 ggacgtagcc aagcgcatga tccgcatggc caaggagtgt ggggctgatt gtgctaagtt 841 ccagaagagt gagctagaat tcaagtttaa tcggaaagcc ttggagaggc catacacctc 901 gaagcattcc tgggggaaga cgtacgggga gcacaaacga catctggagt tcagccatga 961 ccagtacagg gagctgcaga ggtacgccga ggaggttggg atcttcttca ctgcctctgg 1021 catggatgag atggcagttg aattcctgca tgaactgaat gttccatttt tcaaagttgg 1081 atctggagac actaataatt ttccttatct ggaaaagaca gccaaaaaag gtcgcccaat 1141 ggtgatctcc agtgggatgc agtcaatgga caccatgaag caagtttatc agatcgtgaa 1201 gcccctcaac cccaacttct gcttcttgca gtgtaccagc gcatacccgc tccagcctga 1261 ggacgtcaac ctgcgggtca tctcggaata tcagaagctc tttcctgaca ttcccatagg 1321 gtattctggg catgaaacag gcatagcgat atctgtggcc gcagtggctc tgggggccaa 1381 ggtgttggaa cgtcacataa ctttggacaa gacctggaag gggagtgacc actcggcctc 1441 gctggagcct ggagaactgg ccgagctggt gcggtcagtg cgtcttgtgg agcgtgccct 1501 gggctcccca accaagcagc tgctgccctg tgagatggcc tgcaatgaga agctgggcaa 1561 gtctgtggtg gccaaagtga aaattccgga aggcaccatt ctaacaatgg acatgctcac 1621 cgtgaaggtg ggtgagccca aaggctatcc tcctgaagac atctttaatc tagtgggcaa 1681 gaaggtcctg gtcactgttg aagaggatga caccatcatg gaagaattgg tagataatca 1741 tggcaaaaaa atcaagtctg aattctgcag atatccagca cagtggcggc cgctcgagga 1801 ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga 1861 caagtagtga gggcccgata tctcgacaat caacctctgg attacaaaat ttgtgaaaga 1921 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg 1981 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc 2041 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc 2101 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt 2161 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt 2221 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg 2281 aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg 2341 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg 2401 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt 2461 tgggccgcct ccccgcctgg aaacggggga ggctaactga aacacggaag gagacaatac 2521 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac gggtgttggg 2581 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg 2641 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt 2701 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcagat 2761 ctgcgcagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg 2821 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct 2881 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta 2941 aatcggggca tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa 3001 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct 3061 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc 3121 aaccctatct cggtctattc ttttgattta taagggattt tggggatttc ggcctattgg 3181 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc 3241 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc 3301 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc 3361 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc 3421 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt 3481 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt 3541 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg 3601 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt 3661 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct 3721 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga 3781 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg 3841 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact 3901 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg 3961 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct 4021 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg 4081 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt 4141 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg 4201 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc 4261 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag 4321 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt 4381 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt 4441 cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc 4501 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc 4561 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat 4621 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg 4681 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg 4741 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat 4801 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc 4861 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga 4921 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt 4981 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg 5041 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac 5101 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg 5161 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca 5221 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc 5281 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc 5341 ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag 5401 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc 5461 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca 5521 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg 5581 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg 5641 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct 5701 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa 5761 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa 5821 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa 5881 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc 5941 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga 6001 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca 6061 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc 6121 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat 6181 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc 6241 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt 6301 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc 6361 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg 6421 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt 6481 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg 6541 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga 6601 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg 6661 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg 6721 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt 6781 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc 6841 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca 6901 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt 6961 cgactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt 7021 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct 7081 tgaccgacaa ttgcatgaag aatctgctta gg //