LOCUS V029983 7083 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V029983 VERSION V029983 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..1809 /label="Usp18(NM_011909)" /note="Usp18(NM_011909)" /gene="Usp18" CDS 1816..1839 /codon_start=1 /product="8xHis affinity tag" /transl_table=1 /translation="HHHHHHHH" /label="8xHis" misc_feature 1859..2447 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2513..2561 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2763..3191 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3205..3534 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3601..4395 /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 4571..4692 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4741..4757) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4765..4781 /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(4789..4819) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4834..4855 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5143..5731) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5902..6762) /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(6763..6867) /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 caaggggttt 721 gggctcctga ggaaaccctg ccagtcagtt gtggctgagc cgcagcagta ctcagcgctg 781 gaggaagaga ggaccatgaa gaggaagaga gtgctgtcta gagacctctg cagtgcctgg 841 gacagccctc atggtctggt tggtttacac aacatcggac agacgtgttg ccttaactcc 901 ttgcttcaag tgttcatgat gaacatggac ttcagaatga tattgaagag gataacagtg 961 cctcggagtg cagaagagag gaagagaagt gtccccttcc agctgctttt gctgctggag 1021 aagatgcagg acagtcgaca gaaagccgtg ctgcccacgg agctcgttca gtgcctgcag 1081 aaatacaacg tgccgttgtt tgtccagcac gatgctgctc aactctacct tacaatctgg 1141 aacctgacta aggaccagat cacggacaca gacttgacag agcggctgca gggtctgttc 1201 accatctgga cgcaggagtc cctgatttgc gtgggctgta cagcagagag cagcaggagg 1261 agcaaactgc tcacactctc tcttcctctt tttgataagg acgcaaagcc tctgaaaacc 1321 ctggaggatg ccttgagatg tttcgtccag cccaaagagt tagcaagctc cgacatgtgc 1381 tgcgagagct gtggggagaa gacgccttgg aaacaggttc tgaagctgac tcatctgccc 1441 caaaccttga ccattcacct catgagattc tctgccagga actcgaggac ggaaaagatc 1501 tgtcactcag ttaatttccc tcagagcttg gatttcagtc aggttctgcc aaccgaggaa 1561 gacctcggtg ataccaagga acagtctgaa atacactatg aactctttgc cgtgattgcc 1621 cacgttggga tggctgactt tggtcattac tgtgcctaca tccggaatcc cgtggatgga 1681 aagtggttct gcttcaatga ctcacatgtt tgttgggtca cctggaagga tgtccagtgt 1741 acctatggga accacagata ccgctggcgg gaaacggcct atctcttggt ttacacgaag 1801 actggatccc tcgagcacca tcaccatcac catcaccatt agggcccgat atctcgacaa 1861 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc 1921 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat 1981 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg 2041 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg 2101 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat 2161 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt 2221 gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc 2281 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa 2341 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg 2401 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaaacggggg 2461 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata 2521 aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg gggttcggtc 2581 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt 2641 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa 2701 cgtcggggcg gcaggccctg ccatagcaga tctgcgcagc tggggctcta gggggtatcc 2761 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac 2821 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc 2881 cacgttcgcc ggctttcccc gtcaagctct aaatcggggc atccctttag ggttccgatt 2941 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg 3001 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag 3061 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt 3121 ataagggatt ttggggattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt 3181 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc 3241 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag 3301 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc 3361 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct 3421 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct 3481 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc 3541 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc 3601 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc 3661 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca 3721 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg 3781 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg 3841 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag 3901 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg 3961 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc 4021 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa 4081 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac 4141 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat 4201 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac 4261 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc 4321 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt 4381 gacgagttct tctgagcggg actctggggt tcgcgaaatg accgaccaag cgacgcccaa 4441 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat 4501 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt 4561 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac 4621 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat 4681 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg 4741 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc 4801 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc 4861 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat 4921 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 4981 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 5041 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 5101 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 5161 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 5221 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 5281 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg 5341 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 5401 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 5461 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 5521 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 5581 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 5641 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 5701 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 5761 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 5821 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 5881 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat 5941 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg 6001 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc 6061 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc 6121 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc 6181 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc 6241 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc 6301 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa 6361 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat 6421 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata 6481 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca 6541 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag 6601 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc 6661 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc 6721 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata 6781 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta 6841 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga 6901 cggatcggga gatctcccga tcccctatgg tcgactctca gtacaatctg ctctgatgcc 6961 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg 7021 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt 7081 agg //