LOCUS V033576 6098 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V033576 VERSION V033576 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 235..614 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 615..818 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 863..881 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 941..1588 /label="Gstz1(NM_010363)" /note="Gstz1(NM_010363)" /gene="Gstz1" CDS 1595..1660 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 1698..1922 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 1968..2396 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2410..2739 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2806..3600 /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 3774..3895 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(3944..3960) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 3968..3984 /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(3992..4022) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4037..4058 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4346..4931) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5102..5962) /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(5963..6067) /gene="bla" /label="AmpR promoter" ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atgcaagccg ggaagcctat 961 cctctactcc tatttccgaa gctcctgttc atggagagtt cgaattgctc tggcgttaaa 1021 aggcattgac tatgagatag tgcccatcaa cctcataaag gatggcgggc aacagttcac 1081 tgaggaattt cagaccctga accccatgaa gcaagtgcca gcgctgaaaa ttgatggaat 1141 caccattgtc cagtcactgg ccatcatgga gtacctggaa gagactcggc ctatcccacg 1201 gctcctgcct caggacccac agaaaagagc catcgtgcgc atgatttctg acctcatcgc 1261 tagtggcatc cagccccttc agaacctgtc tgtcctgaag caagtgggac aggagaacca 1321 gatgcaatgg gcccaaaagg ttattacttc cggctttaac gctctggaga agattctgca 1381 gagcacagca gggaagtact gcgtaggaga tgaggtatcc atggctgatg tgtgcttagt 1441 accccaagtg gcaaatgctg aaaggttcaa ggtggatcta agcccctacc ctaccatcag 1501 tcacatcaac aaggagctgc tagccttaga ggttttccaa gtgtctcacc cccgtcgaca 1561 gccagatacc cctgccgagc tgaggactct cgaggactac aaagaccatg acggtgatta 1621 taaagatcat gacatcgact acaaggatga cgatgacaag tagtgagggc ccgtttaaac 1681 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc 1741 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga 1801 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga 1861 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat 1921 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag 1981 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag 2041 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt 2101 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca 2161 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata 2221 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca 2281 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc 2341 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt 2401 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt 2461 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca 2521 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta 2581 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga 2641 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag 2701 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata 2761 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat 2821 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca 2881 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg 2941 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg 3001 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact 3061 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct 3121 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg 3181 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt 3241 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc 3301 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc 3361 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga 3421 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc 3481 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt 3541 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga 3601 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt 3661 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg 3721 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt 3781 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag 3841 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg 3901 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg 3961 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta 4021 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg 4081 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga 4141 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg 4201 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag 4261 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc 4321 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca 4381 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt 4441 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc 4501 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc 4561 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc 4621 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact 4681 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg 4741 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta 4801 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca 4861 aacaaaccac cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa 4921 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa 4981 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt 5041 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca 5101 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca 5161 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc 5221 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa 5281 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc 5341 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca 5401 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat 5461 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag 5521 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac 5581 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt 5641 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt 5701 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc 5761 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat 5821 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca 5881 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga 5941 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg 6001 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg 6061 ttccgcgcac atttccccga aaagtgccac ctgacgtc //