LOCUS V030809 6424 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V030809 VERSION V030809 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 964..1914 /label="TAS2R14(NM_023922)" /note="TAS2R14(NM_023922)" /gene="TAS2R14" CDS 1921..1986 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2024..2248 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2294..2722 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2736..3065 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3132..3926 /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 4100..4221 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4270..4286) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4294..4310 /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(4318..4348) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4363..4384 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4672..5257) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5428..6288) /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(6289..6393) /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 accatgggtg gtgtcataaa gagcatattt acattcgttt taattgtgga atttataatt 1021 ggaaatttag gaaatagttt catagcactg gtgaactgta ttgactgggt caagggaaga 1081 aagatctctt cggttgatcg gatcctcact gctttggcaa tctctcgaat tagcctggtt 1141 tggttaatat tcggaagctg gtgtgtgtct gtgtttttcc cagctttatt tgccactgaa 1201 aaaatgttca gaatgcttac taatatctgg acagtgatca atcattttag tgtctggtta 1261 gctacaggcc tcggtacttt ttattttctc aagatagcca atttttctaa ctctattttt 1321 ctctacctaa agtggagggt taaaaaggtg gttttggtgc tgcttcttgt gacttcggtc 1381 ttcttgtttt taaatattgc actgataaac atccatataa atgccagtat caatggatac 1441 agaagaaaca agacttgcag ttctgattca agtaacttta cacgattttc cagtcttatt 1501 gtattaacca gcactgtgtt cattttcata ccctttactt tgtccctggc aatgtttctt 1561 ctcctcatct tctccatgtg gaaacatcgc aagaagatgc agcacactgt caaaatatcc 1621 ggagacgcca gcaccaaagc ccacagagga gttaaaagtg tgatcacttt cttcctactc 1681 tatgccattt tctctctgtc ttttttcata tcagtttgga cctctgaaag gttggaggaa 1741 aatctaatta ttctttccca ggtgatggga atggcttatc cttcatgtca ctcatgtgtt 1801 ctgattcttg gaaacaagaa gctgagacag gcctctctgt cagtgctact gtggctgagg 1861 tacatgttca aagatgggga gccctcaggt cacaaagaat ttagagaatc atctctcgag 1921 gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa ggatgacgat 1981 gacaagtagt gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc 2041 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc 2101 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct 2161 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg 2221 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct 2281 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg 2341 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct 2401 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta 2461 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt 2521 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg 2581 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat 2641 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt 2701 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt 2761 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca 2821 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt 2881 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt 2941 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg 3001 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt 3061 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag 3121 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg 3181 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt 3241 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc 3301 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt 3361 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag 3421 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg 3481 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag 3541 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg 3601 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc 3661 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca 3721 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc 3781 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg 3841 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct 3901 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc 3961 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg 4021 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct 4081 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa 4141 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc 4201 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc 4261 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa 4321 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac 4381 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca 4441 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc 4501 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc 4561 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc 4621 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag 4681 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc 4741 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt 4801 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct 4861 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg 4921 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct 4981 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat 5041 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg 5101 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa 5161 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg 5221 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac 5281 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc 5341 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag 5401 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc 5461 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac 5521 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc 5581 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg 5641 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag 5701 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc 5761 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac 5821 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag 5881 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac 5941 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg 6001 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc 6061 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact 6121 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg 6181 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa 6241 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt 6301 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg 6361 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga 6421 cgtc //