LOCUS V030595 6715 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V030595 VERSION V030595 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..1371 /label="STEEP1(NM_022101)" /note="STEEP1(NM_022101)" /gene="STEEP1" CDS 1378..1467 /codon_start=1 /product="3 tandem Myc epitope tags" /transl_table=1 /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL" /label="3xMyc" misc_feature 1491..2079 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2145..2193 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2395..2823 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2837..3166 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3233..4027 /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 4203..4324 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4373..4389) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4397..4413 /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(4421..4451) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4466..4487 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4775..5363) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5534..6394) /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(6395..6499) /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 ccaccatgcc gaaagtagtg 721 tctcggtcag tagtctgctc tgacactcgg gaccgggagg aatatgacga cggcgagaag 781 cccctccatg tttactactg tttgtgcggc cagatggtcc tagtgctgga ctgccagtta 841 gagaaattgc ccatgaggcc ccgggaccgg tcccgtgtga ttgatgctgc caaacatgcc 901 cataagtttt gtaacacaga agatgaggag actatgtatc tgcggagacc tgaaggcatt 961 gaacgacagt acaggaagaa atgtgcaaag tgtggactgc cgctcttcta ccaatcccag 1021 ccaaagaatg ctcctgttac cttcattgtg gatggagcag tagtcaagtt tggccagggc 1081 tttgggaaaa cgaacatata tactcagaaa caagagcctc ctaagaaggt gatgatgacc 1141 aaacggacca aagacatggg caagttcagt tctgtcaccg tgtctaccat tgatgaagag 1201 gaagaggaga ttgaggctag ggaagttgct gactcatatg cacagaatgc caaagtgatt 1261 gaaaaacagc tggagcgcaa aggcatgagc aagaggcgac tgcaagagct ggctgaattg 1321 gaagccaaga aagcgaaaat gaaggggacc ttgattgaca accagttcaa actcgaggaa 1381 caaaaactaa tatcagagga agatttggaa cagaaactaa tcagtgaaga agatttagag 1441 caaaagttaa tttccgagga ggacttatag tgagggcccg atatctcgac aatcaacctc 1501 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc 1561 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca 1621 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg 1681 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca 1741 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg 1801 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg 1861 acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg 1921 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg 1981 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc 2041 ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaaacggg ggaggctaac 2101 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca 2161 gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct 2221 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct 2281 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg 2341 cggcaggccc tgccatagca gatctgcgca gctggggctc tagggggtat ccccacgcgc 2401 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac 2461 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg 2521 ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt agggttccga tttagtgctt 2581 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc 2641 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct 2701 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga 2761 ttttggggat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga 2821 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg 2881 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg 2941 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc 3001 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca 3061 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt 3121 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc 3181 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga 3241 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga 3301 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg 3361 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga 3421 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt 3481 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct 3541 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct 3601 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg 3661 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca 3721 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga 3781 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt 3841 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt 3901 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct 3961 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt 4021 cttctgagcg ggactctggg gttcgcgaaa tgaccgacca agcgacgccc aacctgccat 4081 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc 4141 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc 4201 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca 4261 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat 4321 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc 4381 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca 4441 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct 4501 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac 4561 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc 4621 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt 4681 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg 4741 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg 4801 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 4861 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 4921 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct 4981 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 5041 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 5101 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 5161 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag 5221 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 5281 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta 5341 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc 5401 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca 5461 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa 5521 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat 5581 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct 5641 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt 5701 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat 5761 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta 5821 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg 5881 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt 5941 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg 6001 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg 6061 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc 6121 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa 6181 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac 6241 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt 6301 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg 6361 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa 6421 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata 6481 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg 6541 gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg ccgcatagtt 6601 aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat 6661 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagg //