LOCUS V018362 9419 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V018362 VERSION V018362 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..4099 /label="MCM9(NM_017696)" /note="MCM9(NM_017696)" /gene="MCM9" CDS 4106..4171 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 4195..4783 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 4849..4897 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 5099..5527 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 5541..5870 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 5937..6731 /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 6907..7028 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(7077..7093) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7101..7117 /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(7125..7155) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7170..7191 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7479..8067) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8238..9098) /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(9099..9203) /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 atgaatagcg atcaagttac actggttggt caagtgtttg agtcatatgt 721 ttcggaatac cataagaatg atattcttct aatcttgaag gaaagggatg aagatgctca 781 ttacccagtt gtggttaatg ccatgactct gtttgagacc aacatggaaa tcggggaata 841 tttcaacatg ttccccagtg aagtgcttac aatttttgat agtgcactgc gaaggtcagc 901 cttgacaatt ctccagtccc tttctcagcc tgaggctgtt tccatgaaac agaatcttca 961 tgccaggata tcaggtttgc ctgtctgtcc tgagctggtg agggaacaca tacctaaaac 1021 caaggatgtg ggacactttt tatctgtcac tgggacagtg attcgaacaa gtctggtgaa 1081 ggttctggag tttgagcggg attacatgtg taacaaatgc aagcatgtgt ttgtgatcaa 1141 ggctgacttt gagcagtatt acaccttttg ccggccatcc tcgtgtccca gcttggagag 1201 ctgtgattcc tctaaattca cttgcctctc aggcttgtct tcgtctccaa ccaggtgtag 1261 agattaccag gaaatcaaaa ttcaggaaca ggttcaaagg ctatctgttg gaagtattcc 1321 acgatctatg aaggttattc tggaagatga cttagtggat agttgcaaat ctggtgatga 1381 cctcactatt tacgggattg taatgcaacg gtggaagccc tttcagcaag atgtgcgctg 1441 tgaagtggag atagtcctga aagcaaatta catccaagta aataatgagc agtcctcagg 1501 gatcatcatg gatgaggagg tccaaaagga attcgaagat ttttgggaat actataagag 1561 cgatcccttt gcaggaagga atgtaatatt ggctagcttg tgccctcaag tgtttggaat 1621 gtatctagta aagcttgctg tggccatggt gctggctggt gggattcaaa ggactgatgc 1681 tacaggaaca cgggtcagag gagaatctca tcttttattg gttggggatc ctggcacagg 1741 gaaatctcag ttcctcaaat atgcagcaaa gattacacca agatctgtgc tgaccacagg 1801 aattggatct actagtgcag gtctgacggt aactgctgta aaagactcag gagaatggaa 1861 tttggaggct ggggcattag ttcttgcaga tgcgggcctt tgctgtattg atgagttcaa 1921 tagcctcaaa gagcatgata ggaccagtat ccatgaagca atggagcaac aaaccataag 1981 tgttgctaag gctggcctcg tgtgcaagct gaacacaagg accaccatcc tggcagcaac 2041 gaaccccaaa ggccagtacg acccccagga gtccgtgtct gtgaacattg ccctcggcag 2101 cccactctta agtcgatttg acctgatcct ggttttgctt gataccaaga atgaagactg 2161 ggatcgtatc atttcctcct ttatcttaga aaataaaggt tacccaagca aatcagagaa 2221 gctctggagc atggaaaaga tgaaaaccta tttctgcctc ataaggaatc tgcagcccac 2281 actgtctgat gtgggcaatc aggttcttct ccggtactac cagatgcaaa ggcagagtga 2341 ttgccggaac gctgcccgga ccaccattcg gctgttggaa agcttgatac gattagcaga 2401 agctcatgct cgcctgatgt ttcgtgatac tgtaactctg gaagacgcta ttacggtggt 2461 gtcagtcatg gagtcctcaa tgcagggagg tgcactgcta ggaggtgtga atgccctcca 2521 cacttccttt cctgaaaacc ctggagagca gtaccagaga cagtgtgaac ttattctgga 2581 aaagctagag ctgcagagcc tcttgagtga agagcttaga agacttgaaa ggttacagaa 2641 tcagagtgtg caccaatccc aaccacgggt attggaggta gagactactc caggatcctt 2701 gagaaatggt ccaggggaag aatcaaactt cagaacttca tcacagcagg aaatcaacta 2761 tagcacacat atcttctctc ctggaggcag ccccgaggga agcccagttc tagatccccc 2821 accgcatctg gagcctaata gatcaacaag taggaaacat tcagctcagc acaaaaataa 2881 cagagatgac agtttagatt ggtttgattt catggcaact catcagagtg aacctaaaaa 2941 cactgttgtt gtgtctcctc atcccaaaac atctggagaa aatatggctt cgaagatctc 3001 taacagcaca tctcagggta aggagaagag tgagccaggc caaaggagca aagtggacat 3061 tgggttgctt ccatcaccag gagagacagg tgttccatgg agggcagaca atgtggaaag 3121 taacaagaaa aaaaggctag cactagattc tgaagcagca gtctctgctg ataaaccaga 3181 ctcagtactg actcatcatg tccccaggaa cctgcagaag ctgtgcaaag agagggccca 3241 gaagttgtgc agaaatagca ccagggtgcc tgcacagtgc acagtccctt cccatcctca 3301 gtccactcct gtacatagcc cagacagaat gctggactca cccaaaagaa agagaccgaa 3361 atcccttgcg caagtggaag agcctgcaat tgaaaatgtt aagcctccag gttcccctgt 3421 ggccaaactg gcaaaattta ctttcaagca gaagtcaaaa ctgatccact cctttgaaga 3481 tcacagccat gtgtcacctg gtgcaactaa aatagcagtt catagtccta aaatttccca 3541 gcgtagaaca agaagagacg cagccttgcc ggtgaagcgt ccaggaaagt taacatctac 3601 cccaggaaac cagatctcca gtcagccaca gggtgagaca aaggaggtgt cgcagcagcc 3661 accagagaaa cacggaccaa gagagaaggt gatgtgtgcc cctgagaaga ggattattca 3721 gcctgaatta gagcttggga acgagactgg gtgtgctcat cttacttgtg agggagacaa 3781 aaaggaagag gtttcaggca gtaataaaag cggcaaggtt catgcctgca cattagccag 3841 attggcaaac ttctgcttta ctcccccatc ggaatccaaa tcaaaatccc ctcctcctga 3901 aaggaagaac cgaggtgaga gaggcccaag ctcccctcct acaaccacag ctccaatgcg 3961 tgtcagtaaa aggaaatctt ttcagctccg tgggtccacc gagaaactga ttgtttccaa 4021 agaatccctc ttcactttac cagaactagg tgatgaagca tttgattgtg actgggatga 4081 agagatgaga aaaaagtcac tcgaggacta caaagaccat gacggtgatt ataaagatca 4141 tgacatcgac tacaaggatg acgatgacaa gtagtgaggg cccgatatct cgacaatcaa 4201 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt 4261 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct 4321 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc 4381 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg 4441 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc 4501 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc 4561 actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt 4621 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca 4681 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt 4741 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaaa cgggggaggc 4801 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa 4861 gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt tcggtcccag 4921 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct 4981 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc 5041 ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg gtatccccac 5101 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct 5161 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg 5221 ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt ccgatttagt 5281 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca 5341 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga 5401 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa 5461 gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac 5521 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag 5581 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc 5641 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag 5701 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc 5761 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc 5821 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg 5881 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga 5941 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct 6001 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc 6061 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg 6121 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg 6181 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc 6241 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc 6301 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg 6361 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc 6421 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg 6481 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc 6541 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag 6601 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg 6661 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg 6721 agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac gcccaacctg 6781 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt 6841 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc 6901 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 6961 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 7021 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca 7081 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga 7141 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg 7201 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc 7261 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac 7321 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata 7381 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa 7441 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct 7501 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa 7561 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg 7621 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca 7681 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa 7741 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg 7801 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg 7861 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg 7921 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc 7981 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag 8041 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac 8101 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc 8161 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag 8221 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt 8281 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag 8341 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca 8401 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact 8461 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca 8521 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg 8581 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc 8641 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg 8701 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca 8761 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt 8821 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc 8881 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc 8941 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca 9001 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa 9061 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat 9121 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa 9181 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga 9241 tcgggagatc tcccgatccc ctatggtcga ctctcagtac aatctgctct gatgccgcat 9301 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca 9361 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttagg //