LOCUS V021192 6413 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V021192 VERSION V021192 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..1060 /label="H2BC13(NM_003519)" /note="H2BC13(NM_003519)" /gene="H2BC13" CDS 1100..1165 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 1189..1777 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 1843..1891 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2093..2521 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2535..2864 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2931..3725 /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 3901..4022 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4071..4087) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4095..4111 /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(4119..4149) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4164..4185 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4473..5061) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5232..6092) /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(6093..6197) /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 ccatgcccga gctggccaag tctgctcccg ccccgaagaa 721 gggctccaag aaggcggtga ccaaggccca gaagaaggat ggcaagaagc gcaagcgcag 781 ccgcaaggag agctactccg tgtacgtgta caaggtgctg aagcaggtcc accccgacac 841 cggcatctct tctaaggcca tgggaatcat gaactccttc gtcaacgaca tcttcgagcg 901 catcgcaagc gaggcttccc gcctggcgca ctacaacaag cgctcgacca tcacctccag 961 ggagatccag accgccgtgc gcctgctgct tccgggggag ctggccaagc acgcggtgtc 1021 ggagggcacc aaggccgtca ccaagtacac cagctccaag gaattctgca gatatccagc 1081 acagtggcgg ccgctcgagg actacaaaga ccatgacggt gattataaag atcatgacat 1141 cgactacaag gatgacgatg acaagtagtg agggcccgat atctcgacaa tcaacctctg 1201 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta 1261 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt 1321 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc 1381 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt 1441 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg 1501 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac 1561 aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc 1621 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac 1681 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct 1741 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaaacggggg aggctaactg 1801 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga 1861 ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg 1921 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt 1981 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg 2041 gcaggccctg ccatagcaga tctgcgcagc tggggctcta gggggtatcc ccacgcgccc 2101 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt 2161 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc 2221 ggctttcccc gtcaagctct aaatcggggc atccctttag ggttccgatt tagtgcttta 2281 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc 2341 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg 2401 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt 2461 ttggggattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat 2521 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca 2581 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct 2641 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc 2701 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg 2761 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc 2821 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt 2881 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac 2941 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact 3001 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc 3061 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg 3121 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg 3181 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt 3241 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc 3301 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag 3361 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg 3421 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc 3481 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt 3541 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg 3601 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt 3661 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct 3721 tctgagcggg actctggggt tcgcgaaatg accgaccaag cgacgcccaa cctgccatca 3781 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg 3841 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc 3901 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca 3961 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct 4021 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg 4081 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata 4141 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca 4201 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc 4261 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg 4321 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta 4381 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc 4441 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag 4501 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac 4561 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc 4621 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt 4681 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc 4741 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga 4801 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta 4861 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta 4921 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga 4981 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg 5041 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag 5101 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc 5161 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact 5221 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt 5281 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta 5341 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta 5401 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc 5461 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat 5521 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt 5581 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg 5641 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca 5701 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta 5761 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg 5821 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact 5881 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg 5941 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt 6001 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga 6061 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc 6121 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa 6181 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga 6241 gatctcccga tcccctatgg tcgactctca gtacaatctg ctctgatgcc gcatagttaa 6301 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt 6361 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agg //