LOCUS V024363 6575 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V024363 VERSION V024363 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..2065 /label="ADH1B(NM_000668)" /note="ADH1B(NM_000668)" /gene="ADH1B" CDS 2072..2137 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2175..2399 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2445..2873 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2887..3216 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3283..4077 /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 4251..4372 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4421..4437) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4445..4461 /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(4469..4499) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4514..4535 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4823..5408) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5579..6439) /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(6440..6544) /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 atgagcacag caggaaaagt 961 aatcaaatgc aaagcagctg tgctatggga ggtaaagaaa cccttttcca ttgaggatgt 1021 ggaggttgca cctcctaagg cttatgaagt tcgcattaag atggtggctg taggaatctg 1081 tcacacagat gaccacgtgg ttagtggcaa cctggtgacc ccccttcctg tgattttagg 1141 ccatgaggca gccggcatcg tggagagtgt tggagaaggg gtgactacag tcaaaccagg 1201 tgataaagtc atcccgctct ttactcctca gtgtggaaaa tgcagagttt gtaaaaaccc 1261 ggagagcaac tactgcttga aaaatgatct aggcaatcct cgggggaccc tgcaggatgg 1321 caccaggagg ttcacctgca gggggaagcc cattcaccac ttccttggca ccagcacctt 1381 ctcccagtac acggtggtgg atgagaatgc agtggccaaa attgatgcag cctcgcccct 1441 ggagaaagtc tgcctcattg gctgtggatt ctcgactggt tatgggtctg cagttaacgt 1501 tgccaaggtc accccaggct ctacctgtgc tgtgtttggc ctgggagggg tcggcctatc 1561 tgctgttatg ggctgtaaag cagctggagc agccagaatc attgcggtgg acatcaacaa 1621 ggacaaattt gcaaaggcca aagagttggg tgccactgaa tgcatcaacc ctcaagacta 1681 caagaaaccc attcaggaag tgctaaagga aatgactgat ggaggtgtgg atttttcgtt 1741 tgaagtcatc ggtcggcttg acaccatgat ggcttccctg ttatgttgtc atgaggcatg 1801 tggcacaagc gtcatcgtag gggtacctcc tgcttcccag aacctctcaa taaaccctat 1861 gctgctactg actggacgca cctggaaggg ggctgtttat ggtggcttta agagtaaaga 1921 aggtatccca aaacttgtgg ctgattttat ggctaagaag ttttcactgg atgcgttaat 1981 aacccatgtt ttaccttttg aaaaaataaa tgaaggattt gacctgcttc actctgggaa 2041 aagtatccgt accgtcctga cgtttctcga ggactacaaa gaccatgacg gtgattataa 2101 agatcatgac atcgactaca aggatgacga tgacaagtag tgagggcccg tttaaacccg 2161 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt 2221 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat 2281 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag 2341 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc 2401 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg 2461 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc 2521 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc 2581 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct 2641 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac 2701 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac 2761 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat 2821 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg 2881 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg 2941 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca 3001 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact 3061 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta 3121 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag 3181 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc 3241 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga 3301 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa 3361 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt 3421 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg 3481 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa 3541 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac 3601 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt 3661 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact 3721 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg 3781 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg 3841 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc 3901 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt 3961 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc 4021 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg 4081 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg 4141 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct 4201 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta 4261 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat 4321 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct 4381 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt 4441 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag 4501 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt 4561 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag 4621 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg 4681 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat 4741 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta 4801 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa 4861 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc 4921 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt 4981 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca 5041 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg 5101 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat 5161 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta 5221 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct 5281 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac 5341 aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag 5401 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact 5461 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa 5521 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt 5581 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag 5641 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca 5701 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc 5761 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt 5821 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg 5881 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca 5941 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg 6001 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca 6061 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg 6121 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct 6181 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca 6241 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca 6301 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg 6361 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac 6421 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt 6481 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc 6541 cgcgcacatt tccccgaaaa gtgccacctg acgtc //