LOCUS V025754 7492 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V025754 VERSION V025754 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 931..2931 /label="NEDD1(NM_001135175)" /note="NEDD1(NM_001135175)" /gene="NEDD1" CDS 2938..2964 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 2968..2994 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 2998..3024 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" polyA_signal 3059..3283 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3329..3757 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3771..4100 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4167..4961 /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 5135..5256 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5305..5321) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5329..5345 /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(5353..5383) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5398..5419 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5707..6292) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6463..7323) /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(7324..7428) /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 cactagtcca gtgtggtgga attcgccacc atgcatttta caggcgcagt catgcaggaa 961 aacctcagat ttgcttcatc aggagatgat attaaaatat gggatgcttc atctatgaca 1021 ttggtggata aattcaaccc acacacatca ccacatggaa tcagctcaat atgttggagc 1081 agcaataata actttttagt aacagcatct tccagtggcg acaaaatagt tgtctcaagt 1141 tgcaaatgta aacctgttcc acttttagag cttgctgaag ggcaaaagca gacatgtgtc 1201 aatttaaatt ctacatctat gtatttggta agcggaggcc taaataacac tgttaatatt 1261 tgggatttaa aatcaaaaag agttcatcga tctcttaagg atcataaaga tcaagtaact 1321 tgtgtaacat acaattggaa tgattgctac attgcttctg gatctcttag tggtgaaatt 1381 attttacaca gtgtaaccac taatttatct agtactcctt ttggccatgg tagtaaccag 1441 tctgttcggc acttgaagta ctccttgttt aagaaatcac tactgggcag tgtttcggat 1501 aatggaatag taactctctg ggatgtaaat agtcagagtc cataccataa ctttgacagt 1561 gtacacaaag ctccagcgtc aggcatctgt ttttctcctg tcaatgaatt gctctttgta 1621 accataggct tggataaaag aatcatcctc tatgacactt caagtaagaa gctagtgaaa 1681 actttagtgg ctgacactcc tctaactgcg gtagatttca tgcctgatgg agccactttg 1741 gctattggat cttcccgggg gaaaatatat caatatgatt taagaatgtt gaaatcacca 1801 gttaagacca tcagtgctca caagacatct gtgcagtgta tagcatttca gtactccact 1861 gttcttacta agtcaagttt aaataaaggc tgttcaaata agcccacaac agtgaacaaa 1921 cgaagtgtta atgtgaatgc tgctagtgga ggagttcaga attccggaat tgtcagagaa 1981 gcacctgcca cgtccattgc cacagttcta ccacaaccta tgacatcagc tatggggaaa 2041 ggaacagttg ctgttcaaga aaaagcaggt ttgcctcgaa gcataaacac agacacttta 2101 tctaaggaaa cagacagtgg aaaaaatcag gatttctcca gctttgatga tactgggaaa 2161 agtagtttag gtgacatgtt ctcacctatc agagatgatg ctgtagttaa caagggaagt 2221 gatgagtcca taggcaaagg agatggcttt gactttctac cgcagttgaa ctcagtgttt 2281 cctccaagaa aaaatccagt aacttcaagt acttcagtat tgcattctag tcctcttaat 2341 gtttttatgg gatctccagg gaaagaggaa aatgaaaacc gtgatctaac agctgagtct 2401 aagaaaatat atatgggaaa acaggaatct aaagactcct tcaaacagtt agcaaagttg 2461 gtcacatctg gtgctgaaag tggaaatcta aatacctctc catcatctaa ccaaacaaga 2521 aattctgaga aatttgaaaa gccagagaat gaaattgaag cccagttgat atgtgaaccc 2581 ccaatcaatg gatcctcaac tccaaatcca aagatagcat cttctgtcac tgctggagtt 2641 gccagttcac tctcagaaaa aatagccgac agcattggaa ataaccggca aaatgcacca 2701 ttgacttcca ttcaaattcg ttttattcag aacatgatac aggaaacgtt ggatgacttt 2761 agagaagcat gccataggga cattgtgaat ttgcaagtgg agatgattaa acagtttcat 2821 atgcaactga atgaaatgca ttctttgctg gaaagatact cagtgaatga aggtttagtg 2881 gctgaaattg aaagactacg agaagaaaac aaaagattac gggcccactt tctcgagtac 2941 ccatacgacg tacctgatta tgcaacttat ccttatgatg tacctgatta tgctagctat 3001 ccttatgatg tcccagacta cgcatgaggg cccgtttaaa cccgctgatc agcctcgact 3061 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg 3121 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg 3181 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg 3241 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga 3301 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg 3361 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct 3421 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat 3481 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt 3541 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg 3601 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac 3661 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta 3721 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt 3781 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca 3841 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa 3901 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc 3961 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg 4021 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg 4081 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc 4141 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc 4201 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct 4261 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg 4321 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca 4381 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc 4441 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga 4501 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc 4561 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc 4621 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg 4681 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct 4741 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc 4801 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc 4861 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc 4921 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga 4981 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt 5041 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg 5101 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg 5161 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc 5221 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc 5281 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct 5341 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg 5401 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct 5461 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg 5521 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc 5581 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 5641 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 5701 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 5761 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 5821 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 5881 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 5941 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 6001 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 6061 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 6121 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc 6181 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 6241 cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 6301 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 6361 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 6421 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 6481 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 6541 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 6601 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 6661 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 6721 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 6781 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 6841 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 6901 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 6961 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 7021 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 7081 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 7141 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 7201 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 7261 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 7321 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 7381 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 7441 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gt //