LOCUS V039342 7435 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V039342 VERSION V039342 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 706..2091 /label="Eef1a1(NM_010106)" /note="Eef1a1(NM_010106)" /gene="Eef1a1" CDS 2098..2187 /codon_start=1 /product="3 tandem Myc epitope tags" /transl_table=1 /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL" /label="3xMyc" misc_feature 2211..2799 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2865..2913 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3115..3543 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3557..3886 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3953..4747 /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 4923..5044 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5093..5109) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5117..5133 /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(5141..5171) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5186..5207 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5495..6083) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6254..7114) /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(7115..7219) /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 gtggaattcg ccaccatggg aaaggaaaag 721 actcacatca acatcgtcgt aatcggacac gtagattccg gcaagtccac cacaaccggc 781 cacctgatct acaaatgtgg tggaatcgac aagcgaacca tcgaaaagtt tgagaaggag 841 gctgctgaga tgggaaaggg ctccttcaag tacgcctggg tcttagacaa actgaaagct 901 gagcgtgagc gtggtatcac tattgacatc tccctgtgga aattcgagac cagcaaatac 961 tatgtgacca tcattgatgc cccaggacac agagacttca tcaaaaacat gattacaggc 1021 acatcccagg ctgactgtgc tgtcctgatt gttgctgctg gtgttggtga atttgaagct 1081 ggtatctcca agaacgggca gacccgcgag catgctcttc tggcttacac cctgggtgtg 1141 aaacagctga ttgttggtgt caacaaaatg gattccaccg agccaccata cagtcagaag 1201 agatacgagg aaatcgttaa ggaagtcagc acctacatta agaaaattgg ctacaaccct 1261 gacacagtag catttgtgcc aatttctggt tggaatggtg acaacatgct ggagccaagt 1321 gctaatatgc cttggttcaa gggatggaaa gtcacccgca aagatggcag tgccagtggc 1381 accacgctgc tggaagcttt ggattgtatc ctaccaccaa ctcgtccaac tgacaagccc 1441 ctgcgactgc ccctccagga tgtctataaa attggaggca ttggcactgt ccctgtgggc 1501 cgagtggaga ctggtgttct caagcctggc atggtggtta cctttgctcc agtcaatgta 1561 acaactgaag tcaagtctgt tgaaatgcac catgaagctt tgagtgaagc tcttcctggg 1621 gacaatgtgg gcttcaatgt aaagaacgtg tcggtcaaag atgttagacg aggcaatgtt 1681 gctggtgaca gcaaaaacga cccaccaatg gaagcagctg gcttcactgc tcaggtgatt 1741 atcctgaacc atccaggcca aatcagtgct ggctacgctc ctgttctgga ttgtcacaca 1801 gcccacatag catgcaagtt tgctgagctt aaagaaaaga tcgatcgtcg ttctggtaag 1861 aagctggaag atggccccaa gttcctgaag tctggcgatg ctgccattgt tgatatggtc 1921 cctggcaagc ccatgtgtgt tgagagcttc tctgactacc ctccacttgg tcgctttgct 1981 gttcgtgaca tgaggcagac agttgctgtg ggtgtcatca aagctgtgga caagaaggct 2041 gctggagctg gcaaagtcac caagtctgcc cagaaagctc agaaggctaa actcgaggaa 2101 caaaaactaa tatcagagga agatttggaa cagaaactaa tcagtgaaga agatttagag 2161 caaaagttaa tttccgagga ggacttatag tgagggcccg atatctcgac aatcaacctc 2221 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc 2281 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca 2341 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg 2401 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca 2461 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg 2521 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg 2581 acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg 2641 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg 2701 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc 2761 ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaaacggg ggaggctaac 2821 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca 2881 gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct 2941 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct 3001 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg 3061 cggcaggccc tgccatagca gatctgcgca gctggggctc tagggggtat ccccacgcgc 3121 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac 3181 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg 3241 ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt agggttccga tttagtgctt 3301 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc 3361 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct 3421 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga 3481 ttttggggat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga 3541 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg 3601 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg 3661 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc 3721 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca 3781 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt 3841 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc 3901 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga 3961 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga 4021 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg 4081 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga 4141 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt 4201 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct 4261 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct 4321 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg 4381 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca 4441 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga 4501 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt 4561 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt 4621 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct 4681 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt 4741 cttctgagcg ggactctggg gttcgcgaaa tgaccgacca agcgacgccc aacctgccat 4801 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc 4861 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc 4921 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca 4981 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat 5041 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc 5101 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca 5161 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct 5221 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac 5281 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc 5341 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt 5401 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg 5461 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg 5521 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 5581 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 5641 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct 5701 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 5761 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 5821 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 5881 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag 5941 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 6001 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta 6061 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc 6121 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca 6181 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa 6241 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat 6301 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct 6361 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt 6421 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat 6481 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta 6541 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg 6601 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt 6661 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg 6721 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg 6781 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc 6841 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa 6901 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac 6961 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt 7021 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg 7081 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa 7141 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata 7201 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg 7261 gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg ccgcatagtt 7321 aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat 7381 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagg //