LOCUS V020369 6605 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V020369 VERSION V020369 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..1261 /label="HPCA(NM_002143)" /note="HPCA(NM_002143)" /gene="HPCA" CDS 1268..1357 /codon_start=1 /product="3 tandem Myc epitope tags" /transl_table=1 /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL" /label="3xMyc" misc_feature 1381..1969 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2035..2083 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2285..2713 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2727..3056 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3123..3917 /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 4093..4214 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4263..4279) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4287..4303 /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(4311..4341) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4356..4377 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4665..5253) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5424..6284) /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(6285..6389) /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 ccatgggcaa gcagaacagc aagctgcggc ccgagatgtt 721 gcaggacctg cgagagaaca cagagttctc agagctggag ctgcaggagt ggtacaaggg 781 cttcctcaag gactgcccca caggaatcct caatgtggat gagttcaaga agatctacgc 841 caacttcttt ccctatggtg acgcctccaa gtttgccgag cacgtcttcc gcacctttga 901 caccaacagc gatggcacca tagactttcg ggagttcatc attgcgctga gcgtgacctc 961 gcgcggccgc ctggagcaga agctcatgtg ggccttcagc atgtatgacc tggacggcaa 1021 cggctacatc agccgggagg agatgctgga gatcgtgcag gccatttaca agatggtttc 1081 gtccgtgatg aagatgccgg aggacgagtc gaccccggaa aagaggactg agaaaatctt 1141 ccgccaaatg gacacaaaca acgacggcaa gctgtccttg gaggagttca tccgcggggc 1201 caaaagcgac ccgtccatcg tgcgtctgct gcagtgcgac cccagcagcg cctcccagtt 1261 cctcgaggaa caaaaactaa tatcagagga agatttggaa cagaaactaa tcagtgaaga 1321 agatttagag caaaagttaa tttccgagga ggacttatag tgagggcccg atatctcgac 1381 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct 1441 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt 1501 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg 1561 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact 1621 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct 1681 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg 1741 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc 1801 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc 1861 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt 1921 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaaacggg 1981 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa 2041 taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg 2101 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc 2161 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc 2221 aacgtcgggg cggcaggccc tgccatagca gatctgcgca gctggggctc tagggggtat 2281 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg 2341 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc 2401 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt agggttccga 2461 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt 2521 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat 2581 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat 2641 ttataaggga ttttggggat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa 2701 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct 2761 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa 2821 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa 2881 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt 2941 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct 3001 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc 3061 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc 3121 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat 3181 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt 3241 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac 3301 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg 3361 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc 3421 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa 3481 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc 3541 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg 3601 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg 3661 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa 3721 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg 3781 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct 3841 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc 3901 ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa tgaccgacca agcgacgccc 3961 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga 4021 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc 4081 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc 4141 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc 4201 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca 4261 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga 4321 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt 4381 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga 4441 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc 4501 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg 4561 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc 4621 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc 4681 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga 4741 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc 4801 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa 4861 tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg 4921 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc 4981 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga 5041 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact 5101 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt 5161 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag 5221 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg 5281 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa 5341 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata 5401 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg 5461 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata 5521 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg 5581 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct 5641 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt 5701 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc 5761 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga 5821 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt 5881 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc 5941 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa 6001 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca 6061 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca 6121 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct 6181 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc 6241 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa 6301 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt 6361 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc 6421 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg 6481 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 6541 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 6601 ttagg //