LOCUS V024179 6665 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V024179 VERSION V024179 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..2155 /label="SAG(NM_000541)" /note="SAG(NM_000541)" /gene="SAG" CDS 2162..2227 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2265..2489 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2535..2963 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2977..3306 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3373..4167 /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 4341..4462 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4511..4527) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4535..4551 /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(4559..4589) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4604..4625 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4913..5498) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5669..6529) /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(6530..6634) /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 atggcagcca gcgggaagac 961 cagcaagtcc gaaccgaacc atgttatctt caagaagatc tcccgggaca aatcggtgac 1021 catctacctg gggaacagag actacataga ccatgtcagc caagtccagc ctgtggatgg 1081 tgtcgtgttg gttgatcctg atcttgtgaa gggaaagaaa gtgtatgtca ctctgacctg 1141 cgccttccgc tatggccaag aggacattga cgtgatcggc ttgaccttcc gcagggacct 1201 gtacttctcc cgggtccagg tgtatcctcc tgtgggggcc gcgagcaccc ccacaaaact 1261 gcaagagagc ctgcttaaaa agctggggag caacacgtac ccctttctcc tgacgtttcc 1321 tgactacttg ccctgttcag tgatgttgca gccagctcca caagattcag ggaagtcctg 1381 tggggttgac tttgaggtca aagcattcgc cacagacagc accgatgccg aagaggacaa 1441 aatccccaag aagagctccg tgcgattact gatccgcaaa gtacagcatg ccccacttga 1501 gatgggtccc cagccccgag ctgaggcggc ctggcagttc ttcatgtctg acaagcccct 1561 gcaccttgcg gtctctctca acaaagagat ctatttccat ggggagccca tccctgtgac 1621 cgtgactgtc accaataaca cagagaagac cgtgaagaag attaaagcat tcgtggaaca 1681 ggtggccaat gtggttctct actcgagtga ttattacgtc aagcccgtgg ctatggagga 1741 agcgcaagaa aaagtgccac caaacagcac tttgaccaag acgctgacgc tgctgccctt 1801 gctggctaac aatcgagaaa ggagaggcat tgccctggat gggaaaatca agcacgagga 1861 cacaaacctt gcctccagca ccatcattaa ggagggcata gaccggaccg tcctgggaat 1921 cctggtgtct taccagatca aggtgaagct cacagtgtca ggctttctgg gagagctcac 1981 ctccagtgaa gtcgccactg aggtcccatt ccgcctcatg caccctcagc ctgaggaccc 2041 agctaaggaa agttatcagg atgcaaattt agtttttgag gagtttgctc gccataatct 2101 gaaagatgca ggagaagctg aggaggggaa gagagacaag aatgacgttg atgagctcga 2161 ggactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga 2221 tgacaagtag tgagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg 2281 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc 2341 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc 2401 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag 2461 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc 2521 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac 2581 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc 2641 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt 2701 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg 2761 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac 2821 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta 2881 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat 2941 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag 3001 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc 3061 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat 3121 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt 3181 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc 3241 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt 3301 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga 3361 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg 3421 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg 3481 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc 3541 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct 3601 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa 3661 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg 3721 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa 3781 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat 3841 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg 3901 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc 3961 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac 4021 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg 4081 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc 4141 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag 4201 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg 4261 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc 4321 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca 4381 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt 4441 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg 4501 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca 4561 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca 4621 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc 4681 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt 4741 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact 4801 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag 4861 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata 4921 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc 4981 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg 5041 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc 5101 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg 5161 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc 5221 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga 5281 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg 5341 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa 5401 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ttttttgttt 5461 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta 5521 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat 5581 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa 5641 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct 5701 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta 5761 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct 5821 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg 5881 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa 5941 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt 6001 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta 6061 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca 6121 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta 6181 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct 6241 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg 6301 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac 6361 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact 6421 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa 6481 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt 6541 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat 6601 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg 6661 acgtc 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