LOCUS V046752 7505 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V046752 VERSION V046752 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 202..581 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 582..785 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 830..848 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 983..1006 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" CDS 1007..1879 /label="TAS2R16(NM_016945)" /note="TAS2R16(NM_016945)" /gene="TAS2R16" CDS 1979..1999 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /transl_table=1 /translation="ENLYFQS" /label="TEV site" CDS 2027..3028 /codon_start=1 /note="In the Tet-Off® system, tTA binds to the Tet-responsive element and stimulates transcription only in the absence of tetracycline or doxycycline." /product="tetracycline-controlled transactivator, comprising a fusion of the tetracycline repressor TetR with the C-terminal activation domain of herpes simplex virus VP16" /transl_table=1 /translation="RLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHVKN KRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTRPT EKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTTDS MPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS,AYSRARTKNNYGSTIE GLLDLPDDDAPEEAGLAAPRLSFLPAGHTRRLSTAPPTDVSLGDELHLDGEDVAMAHAD ALDDFDLDMLGDGDSPGPGFTPHDSAPYGALDMADFEFEQMFTDALGIDEYGG*" /label="tTA" polyA_signal 3072..3296 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3342..3770 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3784..4113 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4180..4974 /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 5148..5269 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5318..5334) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5342..5358 /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(5366..5396) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5411..5432 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5720..6305) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6476..7336) /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(7337..7441) /gene="bla" /label="AmpR promoter" ORIGIN 1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt 61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct 121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg 181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta 241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg 301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc 361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa 421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca 481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta 541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt 601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg 661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca 721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca 781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc 841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc 901 cgccaccatg aagacgatca tcgccctgag ctacatcttc tgcctggtat tcgccgacta 961 caaggacgat gatgacgcca gcgactacaa ggacgacgat gacaagatga tacccatcca 1021 actcactgtc ttcttcatga tcatctatgt gcttgagtcc ttgacaatta ttgtgcagag 1081 cagcctaatt gttgcagtgc tgggcagaga atggctgcaa gtcagaaggc tgatgcctgt 1141 ggacatgatt ctcatcagcc tgggcatctc tcgcttctgt ctacagtggg catcaatgct 1201 gaacaatttt tgctcctatt ttaatttgaa ttatgtactt tgcaacttaa caatcacctg 1261 ggaatttttt aatatcctta cattctggtt aaacagcttg cttaccgtgt tctactgcat 1321 caaggtctct tctttcaccc atcacatctt tctctggctg aggtggagaa ttttgaggtt 1381 gtttccctgg atattactgg gttctctgat gattacttgt gtaacaatca tcccttcagc 1441 tattgggaat tacattcaaa ttcagttact caccatggag catctaccaa gaaacagcac 1501 tgtaactgac aaacttgaaa attttcatca gtatcagttc caggctcata cagttgcatt 1561 ggttattcct ttcatcctgt tcctggcctc caccatcttt ctcatggcat cactgaccaa 1621 gcagatacaa catcatagca ctggtcactg caatccaagc atgaaagcgc gcttcactgc 1681 cctgaggtcc cttgccgtct tatttattgt gtttacctct tactttctaa ccatactcat 1741 caccattata ggtactctat ttgataagag atgttggtta tgggtctggg aagcttttgt 1801 ctatgctttc atcttaatgc attccacttc actgatgctg agcagcccta cgttgaaaag 1861 gattctaaag ggaaagtgcg atatcatcga taccggtgga cgcaccccac ccagcctggg 1921 tccccaagat gagtcctgca ccaccgccag ctcctccctg gccaaggaca cttcatcgga 1981 aaacctgtac ttccaatcca ccggtgagaa cctgtacttc cagctaagat tagataaaag 2041 taaagtgatt aacagcgcat tagagctgct taatgaggtc ggaatcgaag gtttaacaac 2101 ccgtaaactc gcccagaagc taggtgtaga gcagcctaca ttgtattggc atgtaaaaaa 2161 taagcgggct ttgctcgacg ccttagccat tgagatgtta gataggcacc atactcactt 2221 ttgcccttta gaaggggaaa gctggcaaga ttttttacgt aataacgcta aaagttttag 2281 atgtgcttta ctaagtcatc gcgatggagc aaaagtacat ttaggtacac ggcctacaga 2341 aaaacagtat gaaactctcg aaaatcaatt agccttttta tgccaacaag gtttttcact 2401 agagaatgca ttatatgcac tcagcgctgt ggggcatttt actttaggtt gcgtattgga 2461 agatcaagag catcaagtcg ctaaagaaga aagggaaaca cctactactg atagtatgcc 2521 gccattatta cgacaagcta tcgaattatt tgatcaccaa ggtgcagagc cagccttctt 2581 attcggcctt gaattgatca tatgcggatt agaaaaacaa cttaaatgtg aaagtgggtc 2641 cgcgtacagc cgcgcgcgta cgaaaaacaa ttacgggtct accatcgagg gcctgctcga 2701 tctcccggac gacgacgccc ccgaagaggc ggggctggcg gctccgcgcc tgtcctttct 2761 ccccgcggga cacacgcgca gactgtcgac ggcccccccg accgatgtca gcctggggga 2821 cgagctccac ttagacggcg aggacgtggc gatggcgcat gccgacgcgc tagacgattt 2881 cgatctggac atgttggggg acggggattc cccgggtccg ggatttaccc cccacgactc 2941 cgccccctac ggcgctctgg atatggccga cttcgagttt gagcagatgt ttaccgatgc 3001 ccttggaatt gacgagtacg gtgggtagct cgagtctaga gggcccgttt aaacccgctg 3061 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc 3121 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc 3181 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa 3241 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc 3301 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc 3361 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct 3421 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg 3481 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga 3541 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt 3601 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg 3661 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc 3721 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg 3781 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa 3841 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc 3901 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg 3961 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt 4021 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga 4081 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt 4141 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg 4201 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag 4261 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt 4321 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta 4381 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg 4441 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt 4501 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat 4561 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg 4621 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca 4681 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc 4741 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc 4801 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat 4861 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc 4921 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga 4981 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt 5041 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga 5101 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg 5161 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt 5221 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta 5281 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa 5341 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct 5401 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc 5461 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg 5521 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc 5581 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag 5641 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa 5701 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc 5761 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc 5821 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg 5881 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt 5941 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc 6001 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc 6061 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag 6121 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg 6181 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa 6241 ccaccgctgg tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat 6301 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac 6361 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt 6421 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc 6481 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg 6541 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg 6601 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc 6661 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta 6721 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg 6781 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct 6841 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta 6901 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg 6961 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga 7021 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt 7081 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca 7141 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt 7201 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt 7261 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga 7321 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt 7381 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc 7441 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct 7501 atggt 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