LOCUS V040210 7063 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V040210 VERSION V040210 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..2553 /label="Asic3(NM_183000)" /note="Asic3(NM_183000)" /gene="Asic3" CDS 2560..2625 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2663..2887 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2933..3361 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3375..3704 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3771..4565 /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 4739..4860 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4909..4925) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4933..4949 /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(4957..4987) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5002..5023 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5311..5896) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6067..6927) /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(6928..7032) /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 accatgaaac ctccctcagg actggaggag gcccagcggc gacaggcctc agacatccgg 1021 gtgttcgcca acagctgcac gatgcatggt ttgggccaca tctttggccc tggaggcctg 1081 accctgcgcc gtgggctgtg ggccacagct gtactcctgt cgctggcggc cttcctctac 1141 caggtggctg agcgggttcg ctactatggg gagttccacc ataagaccac cctggatgag 1201 cgtgagagcc accagcttac cttcccagct gtcactttgt gtaacatcaa tcccctgcgc 1261 cgctcacgcc tcacacccaa tgacttgcac tgggccggaa cggcactgct gggtctggac 1321 cctgctgaac atgctgccta ccttcgtgcc ctgggccagc cccctgcacc acctggcttc 1381 atgcccagtc cgacttttga catggcacaa ctctacgcca gagccggcca ctcccttgag 1441 gacatgttgc tggactgccg ataccgtggc cagccctgtg gacctgagaa cttcacagtg 1501 attttcactc gaatggggca atgctacacc ttcaactctg gtgcccaggg ggcagagctg 1561 ctcaccactc ctaagggcgg tgctggcaat ggactggaga ttatgctgga tgtacagcag 1621 gaggagtatc tgcccatctg gaaggacatg gaagagaccc catttgaggt ggggatccga 1681 gtgcagatcc acggccagga ggaaccccct gccattgacc agctgggctt cggtgctgcc 1741 ccaggccacc agacttttgt gtcctgccag caacagcaac tgagtttcct gccaccaccc 1801 tggggtgact gcaataccgc atctgtggat cccgactttg atccagagcc ctctgatccc 1861 ctgggttccc ctagctccag ccctccttat agcttaatag ggtgtcgcct ggcctgtgag 1921 tcacgctatg tggctcggaa gtgcggatgt cgaatgatgc atatgcctgg aaactcccca 1981 gtgtgcagcc cccagcagta caaggactgt gccagcccag ctctggacgc tatgctgcga 2041 aaggacactt gtgtctgtcc caacccgtgc gccactacac gctatgccaa ggagctctcc 2101 atggtgcgga ttcccagccg cgcttcagct cgctacctgg cccggaaata caaccgtagc 2161 gagacttaca tcacggagaa tgtactggtt ctggatatct tctttgaagc cctcaactat 2221 gaggccgtgg aacaaaaggc agcttatgaa gtgtcggagt tgctgggaga cattggggga 2281 cagatgggac tgtttatcgg agccagcctg cttaccatcc tcgagatcct cgactacctc 2341 tgtgaggttt ttcaagacag agtcctgggg tacttctgga acagaaggag ctctcaaagg 2401 cgctctggca acactctgct ccaggaagag ttgaatgggc atcgaacaca tgttccccat 2461 ctcagcctag gccccaggcc tcctaccgct cccagtgctg tcaccaagac actcgctgcc 2521 tcccaccgta cctgttacct cgtgacaagg ctcctcgagg actacaaaga ccatgacggt 2581 gattataaag atcatgacat cgactacaag gatgacgatg acaagtagtg agggcccgtt 2641 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc 2701 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat 2761 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg 2821 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc 2881 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc 2941 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt 3001 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc 3061 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta 3121 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc 3181 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg 3241 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt 3301 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat 3361 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca 3421 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct 3481 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc 3541 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg 3601 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc 3661 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt 3721 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac 3781 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact 3841 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc 3901 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg 3961 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg 4021 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt 4081 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc 4141 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag 4201 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg 4261 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc 4321 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt 4381 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg 4441 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt 4501 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct 4561 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg 4621 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga 4681 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa 4741 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa 4801 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta 4861 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt 4921 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa 4981 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact 5041 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc 5101 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg 5161 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc 5221 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag 5281 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca 5341 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca 5401 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg 5461 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag 5521 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt 5581 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca 5641 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg 5701 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt 5761 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc 5821 cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag 5881 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 5941 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 6001 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 6061 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 6121 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 6181 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 6241 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 6301 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 6361 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 6421 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 6481 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 6541 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 6601 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 6661 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 6721 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 6781 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 6841 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 6901 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta 6961 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 7021 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc //