LOCUS V030622 6376 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V030622 VERSION V030622 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 970..1866 /label="TAS2R1(NM_019599)" /note="TAS2R1(NM_019599)" /gene="TAS2R1" CDS 1873..1938 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 1976..2200 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2246..2674 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2688..3017 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3084..3878 /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 4052..4173 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4222..4238) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4246..4262 /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(4270..4300) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4315..4336 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4624..5209) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5380..6240) /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(6241..6345) /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 atccactagt ccagtgtggt ggaattcgcc 961 accgccacca tgctagagtc tcacctcatt atctattttc ttcttgcagt gatacaattt 1021 cttcttggga ttttcacaaa tggcatcatt gtggtggtga atggcattga cttgatcaag 1081 cacagaaaaa tggctccgct ggatctcctt ctttcttgtc tggcagtttc tagaattttt 1141 ctgcagttgt tcatcttcta cgttaatgtg attgttatct tcttcataga attcatcatg 1201 tgttctgcga attgtgcaat tctcttattt ataaatgaat tggaactttg gcttgccaca 1261 tggctcggcg ttttctattg tgccaaggtt gccagcgtcc gtcacccact cttcatctgg 1321 ttgaagatga ggatatccaa gctggtccca tggatgatcc tggggtctct gctatatgta 1381 tctatgattt gtgttttcca tagcaaatat gcagggttta tggtcccata cttcctaagg 1441 aaatttttct cccaaaatgc cacaattcaa aaagaagata cactggctat acagattttc 1501 tcttttgttg ctgagttctc agtgccattg cttatcttcc tttttgctgt tttgctcttg 1561 attttctctc tggggaggca cacccggcaa atgagaaaca cagtggccgg cagcagggtt 1621 cctggcaggg gtgcacccat cagcgcgttg ctgtctatcc tgtccttcct gatcctctac 1681 ttctcccact gcatgataaa agtttttctc tcttctctaa agtttcacat cagaaggttc 1741 atctttctgt tcttcatcct tgtgattggt atataccctt ctggacactc tctcatctta 1801 attttaggaa atcctaaatt gaaacaaaat gcaaaaaagt tcctcctcca cagtaagtgc 1861 tgtcagctcg aggactacaa agaccatgac ggtgattata aagatcatga catcgactac 1921 aaggatgacg atgacaagta gtgagggccc gtttaaaccc gctgatcagc ctcgactgtg 1981 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa 2041 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt 2101 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa 2161 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc 2221 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt 2281 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc 2341 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg 2401 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat 2461 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg 2521 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct 2581 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa 2641 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag 2701 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt 2761 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca 2821 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa 2881 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag 2941 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag 3001 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag 3061 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg 3121 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg 3181 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc 3241 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga 3301 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc 3361 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag 3421 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat 3481 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg 3541 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca 3601 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct 3661 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg 3721 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg 3781 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc 3841 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat 3901 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta 3961 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg 4021 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta 4081 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag 4141 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag 4201 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac 4261 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 4321 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 4381 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 4441 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 4501 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 4561 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 4621 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 4681 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 4741 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 4801 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 4861 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 4921 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 4981 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 5041 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt 5101 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 5161 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 5221 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 5281 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 5341 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 5401 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 5461 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 5521 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 5581 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 5641 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 5701 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 5761 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 5821 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 5881 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 5941 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 6001 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 6061 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 6121 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 6181 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 6241 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 6301 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 6361 agtgccacct gacgtc //