LOCUS V024158 7079 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V024158 VERSION V024158 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..2569 /label="YES1(NM_005433)" /note="YES1(NM_005433)" /gene="YES1" CDS 2576..2641 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2679..2903 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2949..3377 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3391..3720 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3787..4581 /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 4755..4876 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4925..4941) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4949..4965 /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(4973..5003) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5018..5039 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5327..5912) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6083..6943) /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(6944..7048) /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 atgggctgca ttaaaagtaa 961 agaaaacaaa agtccagcca ttaaatacag acctgaaaat actccagagc ctgtcagtac 1021 aagtgtgagc cattatggag cagaacccac tacagtgtca ccatgtccgt catcttcagc 1081 aaagggaaca gcagttaatt tcagcagtct ttccatgaca ccatttggag gatcctcagg 1141 ggtaacgcct tttggaggtg catcttcctc attttcagtg gtgccaagtt catatcctgc 1201 tggtttaaca ggtggtgtta ctatatttgt ggccttatat gattatgaag ctagaactac 1261 agaagacctt tcatttaaga agggtgaaag atttcaaata attaacaata cggaaggaga 1321 ttggtgggaa gcaagatcaa tcgctacagg aaagaatggt tatatcccga gcaattatgt 1381 agcgcctgca gattccattc aggcagaaga atggtatttt ggcaaaatgg ggagaaaaga 1441 tgctgaaaga ttacttttga atcctggaaa tcaacgaggt attttcttag taagagagag 1501 tgaaacaact aaaggtgctt attccctttc tattcgtgat tgggatgaga taaggggtga 1561 caatgtgaaa cactacaaaa ttaggaaact tgacaatggt ggatactata tcacaaccag 1621 agcacaattt gatactctgc agaaattggt gaaacactac acagaacatg ctgatggttt 1681 atgccacaag ttgacaactg tgtgtccaac tgtgaaacct cagactcaag gtctagcaaa 1741 agatgcttgg gaaatccctc gagaatcttt gcgactagag gttaaactag gacaaggatg 1801 tttcggcgaa gtgtggatgg gaacatggaa tggaaccacg aaagtagcaa tcaaaacact 1861 aaaaccaggt acaatgatgc cagaagcttt ccttcaagaa gctcagataa tgaaaaaatt 1921 aagacatgat aaacttgttc cactatatgc tgttgtttct gaagaaccaa tttacattgt 1981 cactgaattt atgtcaaaag gaagcttatt agatttcctt aaggaaggag atggaaagta 2041 tttgaagctt ccacagctgg ttgatatggc tgctcagatt gctgatggta tggcatatat 2101 tgaaagaatg aactatattc accgagatct tcgggctgct aatattcttg taggagaaaa 2161 tcttgtgtgc aaaatagcag actttggttt agcaaggtta attgaagaca atgaatacac 2221 agcaagacaa ggtgcaaaat ttccaatcaa atggacagct cctgaagctg cactgtatgg 2281 tcggtttaca ataaagtctg atgtctggtc atttggaatt ctgcaaacag aactagtaac 2341 aaagggccga gtgccatatc caggtatggt gaaccgtgaa gtattagaac aagtggagcg 2401 aggatacagg atgccgtgcc ctcagggctg tccagaatcc ctccatgaat tgatgaatct 2461 gtgttggaag aaggaccctg atgaaagacc aacatttgaa tatattcagt ccttcttgga 2521 agactacttc actgctacag agccacagta ccagccagga gaaaatttac tcgaggacta 2581 caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa 2641 gtagtgaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc 2701 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt 2761 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct 2821 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc 2881 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg 2941 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag 3001 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt 3061 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt 3121 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg 3181 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt 3241 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt 3301 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca 3361 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca 3421 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt 3481 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca 3541 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc 3601 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct 3661 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa 3721 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg 3781 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg 3841 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg 3901 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat 3961 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca 4021 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg 4081 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat 4141 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa 4201 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg 4261 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg 4321 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg 4381 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat 4441 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac 4501 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc 4561 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc 4621 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg 4681 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt 4741 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca 4801 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac 4861 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat 4921 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac 4981 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa 5041 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat 5101 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc 5161 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg 5221 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag 5281 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc 5341 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag 5401 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga 5461 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc 5521 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg 5581 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt 5641 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca 5701 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca 5761 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag 5821 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc 5881 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt 5941 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa 6001 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat 6061 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga 6121 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac 6181 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg 6241 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg 6301 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt 6361 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct 6421 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat 6481 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta 6541 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca 6601 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat 6661 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac 6721 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa 6781 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt 6841 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg 6901 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat 6961 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt 7021 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc //