LOCUS V031752 6206 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V031752 VERSION V031752 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 944..961 /codon_start=1 /product="6xHis affinity tag" /transl_table=1 /translation="HHHHHH" /label="6xHis" CDS 962..1696 /label="YWHAZ(NM_003406)" /note="YWHAZ(NM_003406)" /gene="YWHAZ" CDS 1703..1768 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 1806..2030 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2076..2504 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2518..2847 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2914..3708 /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 3882..4003 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4052..4068) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4076..4092 /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(4100..4130) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4145..4166 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4454..5039) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5210..6070) /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(6071..6175) /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 atgcatcacc atcaccatca 961 catggataaa aatgagctgg ttcagaaggc caaactggcc gagcaggctg agcgatatga 1021 tgacatggca gcctgcatga agtctgtaac tgagcaagga gctgaattat ccaatgagga 1081 gaggaatctt ctctcagttg cttataaaaa tgttgtagga gcccgtaggt catcttggag 1141 ggtcgtctca agtattgaac aaaagacgga aggtgctgag aaaaaacagc agatggctcg 1201 agaatacaga gagaaaattg agacggagct aagagatatc tgcaatgatg tactgtctct 1261 tttggaaaag ttcttgatcc ccaatgcttc acaagcagag agcaaagtct tctatttgaa 1321 aatgaaagga gattactacc gttacttggc tgaggttgcc gctggtgatg acaagaaagg 1381 gattgtcgat cagtcacaac aagcatacca agaagctttt gaaatcagca aaaaggaaat 1441 gcaaccaaca catcctatca gactgggtct ggcccttaac ttctctgtgt tctattatga 1501 gattctgaac tccccagaga aagcctgctc tcttgcaaag acagcttttg atgaagccat 1561 tgctgaactt gatacattaa gtgaagagtc atacaaagac agcacgctaa taatgcaatt 1621 actgagagac aacttgacat tgtggacatc ggatacccaa ggagacgaag ctgaagcagg 1681 agaaggaggg gaaaatctcg aggactacaa agaccatgac ggtgattata aagatcatga 1741 catcgactac aaggatgacg atgacaagta gtgagggccc gtttaaaccc gctgatcagc 1801 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt 1861 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca 1921 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga 1981 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc 2041 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag 2101 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc 2161 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc 2221 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa 2281 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg 2341 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac 2401 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta 2461 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg 2521 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg 2581 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt 2641 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc 2701 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt 2761 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc 2821 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga 2881 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca 2941 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc 3001 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc 3061 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg 3121 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg 3181 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct 3241 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct 3301 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa 3361 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa 3421 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc 3481 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt 3541 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct 3601 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc 3661 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg 3721 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg 3781 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc 3841 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt 3901 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac 3961 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt 4021 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt 4081 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg 4141 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg 4201 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 4261 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 4321 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 4381 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 4441 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 4501 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 4561 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 4621 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 4681 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 4741 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 4801 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 4861 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc 4921 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 4981 ctggtagcgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag 5041 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag 5101 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat 5161 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct 5221 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac 5281 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa 5341 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg 5401 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt 5461 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca 5521 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt 5581 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct 5641 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg 5701 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg 5761 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg 5821 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa 5881 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt 5941 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt 6001 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt 6061 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca 6121 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat 6181 ttccccgaaa agtgccacct gacgtc //