LOCUS V025875 7921 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V025875 VERSION V025875 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..2577 /label="KEAP1(NM_012289)" /note="KEAP1(NM_012289)" /gene="KEAP1" CDS 2584..2673 /codon_start=1 /product="3 tandem Myc epitope tags" /transl_table=1 /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL" /label="3xMyc" misc_feature 2697..3285 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 3351..3399 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3601..4029 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4043..4372 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4439..5233 /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 5409..5530 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5579..5595) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5603..5619 /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(5627..5657) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5672..5693 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5981..6569) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6740..7600) /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(7601..7705) /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 ccaccatgca gccagatccc 721 aggcctagcg gggctggggc ctgctgccga ttcctgcccc tgcagtcaca gtgccctgag 781 ggggcagggg acgcggtgat gtacgcctcc actgagtgca aggcggaggt gacgccctcc 841 cagcatggca accgcacctt cagctacacc ctggaggatc ataccaagca ggcctttggc 901 atcatgaacg agctgcggct cagccagcag ctgtgtgacg tcacactgca ggtcaagtac 961 caggatgcac cggccgccca gttcatggcc cacaaggtgg tgctggcctc atccagccct 1021 gtcttcaagg ccatgttcac caacgggctg cgggagcagg gcatggaggt ggtgtccatt 1081 gagggtatcc accccaaggt catggagcgc ctcattgaat tcgcctacac ggcctccatc 1141 tccatgggcg agaagtgtgt cctccacgtc atgaacggtg ctgtcatgta ccagatcgac 1201 agcgttgtcc gtgcctgcag tgacttcctg gtgcagcagc tggaccccag caatgccatc 1261 ggcatcgcca acttcgctga gcagattggc tgtgtggagt tgcaccagcg tgcccgggag 1321 tacatctaca tgcattttgg ggaggtggcc aagcaagagg agttcttcaa cctgtcccac 1381 tgccaactgg tgaccctcat cagccgggac gacctgaacg tgcgctgcga gtccgaggtc 1441 ttccacgcct gcatcaactg ggtcaagtac gactgcgaac agcgacggtt ctacgtccag 1501 gcgctgctgc gggccgtgcg ctgccactcg ttgacgccga acttcctgca gatgcagctg 1561 cagaagtgcg agatcctgca gtccgactcc cgctgcaagg actacctggt caagatcttc 1621 gaggagctca ccctgcacaa gcccacgcag gtgatgccct gccgggcgcc caaggtgggc 1681 cgcctgatct acaccgcggg cggctacttc cgacagtcgc tcagctacct ggaggcttac 1741 aaccccagtg acggcacctg gctccggttg gcggacctgc aggtgccgcg gagcggcctg 1801 gccggctgcg tggtgggcgg gctgttgtac gccgtgggcg gcaggaacaa ctcgcccgac 1861 ggcaacaccg actccagcgc cctggactgt tacaacccca tgaccaatca gtggtcgccc 1921 tgcgccccca tgagcgtgcc ccgtaaccgc atcggggtgg gggtcatcga tggccacatc 1981 tatgccgtcg gcggctccca cggctgcatc caccacaaca gtgtggagag gtatgagcca 2041 gagcgggatg agtggcactt ggtggcccca atgctgacac gaaggatcgg ggtgggcgtg 2101 gctgtcctca atcgtctcct ttatgccgtg gggggctttg acgggacaaa ccgccttaat 2161 tcagctgagt gttactaccc agagaggaac gagtggcgaa tgatcacagc aatgaacacc 2221 atccgaagcg gggcaggcgt ctgcgtcctg cacaactgta tctatgctgc tgggggctat 2281 gatggtcagg accagctgaa cagcgtggag cgctacgatg tggaaacaga gacgtggact 2341 ttcgtagccc ccatgaagca ccggcgaagt gccctgggga tcactgtcca ccaggggaga 2401 atctacgtcc ttggaggcta tgatggtcac acgttcctgg acagtgtgga gtgttacgac 2461 ccagatacag acacctggag cgaggtgacc cgaatgacat cgggccggag tggggtgggc 2521 gtggctgtca ccatggagcc ctgccggaag cagattgacc agcagaactg tacctgtctc 2581 gaggaacaaa aactaatatc agaggaagat ttggaacaga aactaatcag tgaagaagat 2641 ttagagcaaa agttaatttc cgaggaggac ttatagtgag ggcccgatat ctcgacaatc 2701 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt 2761 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg 2821 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc 2881 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt 2941 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg 3001 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg 3061 gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct 3121 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc 3181 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc 3241 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga aacgggggag 3301 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa 3361 aagacagaat aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg gttcggtccc 3421 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt 3481 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg 3541 tcggggcggc aggccctgcc atagcagatc tgcgcagctg gggctctagg gggtatcccc 3601 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg 3661 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca 3721 cgttcgccgg ctttccccgt caagctctaa atcggggcat ccctttaggg ttccgattta 3781 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc 3841 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg 3901 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat 3961 aagggatttt ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta 4021 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc 4081 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc 4141 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat 4201 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc 4261 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga 4321 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc 4381 gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat 4441 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg 4501 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc 4561 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca 4621 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct 4681 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga 4741 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg 4801 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat 4861 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga 4921 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg 4981 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg 5041 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat 5101 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct 5161 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga 5221 cgagttcttc tgagcgggac tctggggttc gcgaaatgac cgaccaagcg acgcccaacc 5281 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg 5341 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg 5401 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa 5461 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca 5521 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt 5581 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg 5641 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt 5701 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg 5761 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg 5821 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa 5881 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc 5941 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc 6001 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 6061 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc 6121 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct 6181 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 6241 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 6301 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga 6361 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa 6421 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta 6481 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc 6541 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 6601 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 6661 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 6721 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct 6781 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg 6841 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 6901 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa 6961 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 7021 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt 7081 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 7141 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt 7201 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 7261 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt 7321 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata 7381 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga 7441 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag 7501 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa 7561 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt 7621 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga 7681 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg 7741 gatcgggaga tctcccgatc ccctatggtc gactctcagt acaatctgct ctgatgccgc 7801 atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag 7861 caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag 7921 g //