LOCUS V019377 7742 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V019377 VERSION V019377 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..3232 /label="TSHR(NM_000369)" /note="TSHR(NM_000369)" /gene="TSHR" CDS 3239..3304 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 3342..3566 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3612..4040 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4054..4383 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4450..5244 /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 5418..5539 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5588..5604) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5612..5628 /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(5636..5666) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5681..5702 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5990..6575) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6746..7606) /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(7607..7711) /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 atgaggccgg cggacttgct 961 gcagctggtg ctgctgctcg acctgcccag ggacctgggc ggaatggggt gttcgtctcc 1021 accctgcgag tgccatcagg aggaggactt cagagtcacc tgcaaggata ttcaacgcat 1081 ccccagctta ccgcccagta cgcagactct gaagcttatt gagactcacc tgagaactat 1141 tccaagtcat gcattttcta atctgcccaa tatttccaga atctacgtat ctatagatgt 1201 gactctgcag cagctggaat cacactcctt ctacaatttg agtaaagtga ctcacataga 1261 aattcggaat accaggaact taacttacat agaccctgat gccctcaaag agctccccct 1321 cctaaagttc cttggcattt tcaacactgg acttaaaatg ttccctgacc tgaccaaagt 1381 ttattccact gatatattct ttatacttga aattacagac aacccttaca tgacgtcaat 1441 ccctgtgaat gcttttcagg gactatgcaa tgaaaccttg acactgaagc tgtacaacaa 1501 tggctttact tcagtccaag gatatgcttt caatgggaca aagctggatg ctgtttacct 1561 aaacaagaat aaatacctga cagttattga caaagatgca tttggaggag tatacagtgg 1621 accaagcttg ctggacgtgt ctcaaaccag tgtcactgcc cttccatcca aaggcctgga 1681 gcacctgaag gaactgatag caagaaacac ctggactctt aagaaacttc cactttcctt 1741 gagtttcctt cacctcacac gggctgacct ttcttaccca agccactgct gtgcttttaa 1801 gaatcagaag aaaatcagag gaatccttga gtccttgatg tgtaatgaga gcagtatgca 1861 gagcttgcgc cagagaaaat ctgtgaatgc cttgaatagc cccctccacc aggaatatga 1921 agagaatctg ggtgacagca ttgttgggta caaggaaaag tccaagttcc aggatactca 1981 taacaacgct cattattacg tcttctttga agaacaagag gatgagatca ttggttttgg 2041 ccaggagctc aaaaaccccc aggaagagac tctacaagct tttgacagcc attatgacta 2101 caccatatgt ggggacagtg aagacatggt gtgtaccccc aagtccgatg agttcaaccc 2161 gtgtgaagac ataatgggct acaagttcct gagaattgtg gtgtggttcg ttagtctgct 2221 ggctctcctg ggcaatgtct ttgtcctgct tattctcctc accagccact acaaactgaa 2281 cgtcccccgc tttctcatgt gcaacctggc ctttgcggat ttctgcatgg ggatgtacct 2341 gctcctcatc gcctctgtag acctctacac tcactctgag tactacaacc atgccatcga 2401 ctggcagaca ggccctgggt gcaacacggc tggtttcttc actgtctttg caagcgagtt 2461 atcggtgtat acgctgacgg tcatcaccct ggagcgctgg tatgccatca ccttcgccat 2521 gcgcctggac cggaagatcc gcctcaggca cgcatgtgcc atcatggttg ggggctgggt 2581 ttgctgcttc cttctcgccc tgcttccttt ggtgggaata agtagctatg ccaaagtcag 2641 tatctgcctg cccatggaca ccgagacccc tcttgctctg gcatatattg tttttgttct 2701 gacgctcaac atagttgcct tcgtcatcgt ctgctgctgt tatgtgaaga tctacatcac 2761 agtccgaaat ccgcagtaca acccagggga caaagatacc aaaattgcca agaggatggc 2821 tgtgttgatc ttcaccgact tcatatgcat ggccccaatc tcattctatg ctctgtcagc 2881 aattctgaac aagcctctca tcactgttag caactccaaa atcttgctgg tactcttcta 2941 tccacttaac tcctgtgcca atccattcct ctatgctatt ttcaccaagg ccttccagag 3001 ggatgtgttc atcctactca gcaagtttgg catctgtaaa cgccaggctc aggcataccg 3061 ggggcagagg gttcctccaa agaacagcac tgatattcag gttcaaaagg ttacccacga 3121 gatgaggcag ggtctccaca acatggaaga tgtctatgaa ctgattgaaa actcccatct 3181 aaccccaaag aagcaaggcc aaatctcaga agagtatatg caaacggttt tgctcgagga 3241 ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga 3301 caagtagtga gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca 3361 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac 3421 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat 3481 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca 3541 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag 3601 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg 3661 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc 3721 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg 3781 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc 3841 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt 3901 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc 3961 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta 4021 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc 4081 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg 4141 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag 4201 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc 4261 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc 4321 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc 4381 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga 4441 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag 4501 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc 4561 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg 4621 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc 4681 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg 4741 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct 4801 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg 4861 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat 4921 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc 4981 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg 5041 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc 5101 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct 5161 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat 5221 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga 5281 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct 5341 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg 5401 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata 5461 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca 5521 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt 5581 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca 5641 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat 5701 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt 5761 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct 5821 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa 5881 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa 5941 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc 6001 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga 6061 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc 6121 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt 6181 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct 6241 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg 6301 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta 6361 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct 6421 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa 6481 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca 6541 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg 6601 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa 6661 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta 6721 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag 6781 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga 6841 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac 6901 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc 6961 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta 7021 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac 7081 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat 7141 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa 7201 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg 7261 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag 7321 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc 7381 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct 7441 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat 7501 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg 7561 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc 7621 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta 7681 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg 7741 tc //