LOCUS V040715 7628 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V040715 VERSION V040715 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 908..3151 /codon_start=1 /gene="" /note="homolog of budding yeast Sir2" /product="human NAD+-dependent protein deacetylase sirtuin-1" /protein_id="" /transl_table=1 /translation="MADEAALALQPGGSPSAAGADREAASSPAGEPLRKRPRRDGPGLE RSPGEPGGAAPEREVPAAARGCPGAAAAALWREAEAEAAAAGGEQEAQATAAAGEGDNG PGLQGPSREPPLADNLYDEDDDDEGEEEEEAAAAAIGYRDNLLFGDEIITNGFHSCESD EEDRASHASSSDWTPRPRIGPYTFVQQHLMIGTDPRTILKDLLPETIPPPELDDMTLWQ IVINILSEPPKRKKRKDINTIEDAVKLLQECKKIIVLTGAGVSVSCGIPDFRSRDGIYA RLAVDFPDLPDPQAMFDIEYFRKDPRPFFKFAKEIYPGQFQPSLCHKFIALSDKEGKLL RNYTQNIDTLEQVAGIQRIIQCHGSFATASCLICKYKVDCEAVRGDIFNQVVPRCPRCP ADEPLAIMKPEIVFFGENLPEQFHRAMKYDKDEVDLLIVIGSSLKVRPVALIPSSIPHE VPQILINREPLPHLHFDVELLGDCDVIINELCHRLGGEYAKLCCNPVKLSEITEKPPRT QKELAYLSELPPTPLHVSEDSSSPERTSPPDSSVIVTLLDQAAKSNDDLDVSESKGCME EKPQEVQTSRNVESIAEQMENPDLKNVGSSTGEKNERTSVAGTVRKCWPNRVAKEQISR RLDGNQYLFLPPNRYIFHGAEVYSDSEDDVLSSSSCGSNSDSGTCQSPSLEEPMEDESE IEEFYNGLEDEPDVPERAGGAGFGTDGDDQEAINEAISVKQEVTDMNYPSNKS*" /label="SIRT1" polyA_signal 3195..3419 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3465..3893 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3907..4236 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4303..5097 /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 5271..5392 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5441..5457) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5465..5481 /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(5489..5519) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5534..5555 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5843..6428) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6599..7459) /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(7460..7564) /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 cgccaccatg gcggacgagg cggccctcgc ccttcagccc ggcggctccc cctcggcggc 961 gggggccgac agggaggccg cgtcgtcccc cgccggggag ccgctccgca agaggccgcg 1021 gagagatggt cccggcctcg agcggagccc gggcgagccc ggtggggcgg ccccagagcg 1081 tgaggtgccg gcggcggcca ggggctgccc gggtgcggcg gcggcggcgc tgtggcggga 1141 ggcggaggca gaggcggcgg cggcaggcgg ggagcaagag gcccaggcga ctgcggcggc 1201 tggggaagga gacaatgggc cgggcctgca gggcccatct cgggagccac cgctggccga 1261 caacttgtac gacgaagacg acgacgacga gggcgaggag gaggaagagg cggcggcggc 1321 ggcgattggg taccgagata accttctgtt cggtgatgaa attatcacta atggttttca 1381 ttcctgtgaa agtgatgagg aggatagagc ctcacatgca agctctagtg actggactcc 1441 aaggccacgg ataggtccat atacttttgt tcagcaacat cttatgattg gcacagatcc 1501 tcgaacaatt cttaaagatt tattgccgga aacaatacct ccacctgagt tggatgatat 1561 gacactgtgg cagattgtta ttaatatcct ttcagaacca ccaaaaagga aaaaaagaaa 1621 agatattaat acaattgaag atgctgtgaa attactgcaa gagtgcaaaa aaattatagt 1681 tctaactgga gctggggtgt ctgtttcatg tggaatacct gacttcaggt caagggatgg 1741 tatttatgct cgccttgctg tagacttccc agatcttcca gatcctcaag cgatgtttga 1801 tattgaatat ttcagaaaag atccaagacc attcttcaag tttgcaaagg aaatatatcc 1861 tggacaattc cagccatctc tctgtcacaa attcatagcc ttgtcagata aggaaggaaa 1921 actacttcgc aactataccc agaacataga cacgctggaa caggttgcgg gaatccaaag 1981 gataattcag tgtcatggtt cctttgcaac agcatcttgc ctgatttgta aatacaaagt 2041 tgactgtgaa gctgtacgag gagatatttt taatcaggta gttcctcgat gtcctaggtg 2101 cccagctgat gaaccgcttg ctatcatgaa accagagatt gtgttttttg gtgaaaattt 2161 accagaacag tttcatagag ccatgaagta tgacaaagat gaagttgacc tcctcattgt 2221 tattgggtct tccctcaaag taagaccagt agcactaatt ccaagttcca taccccatga 2281 agtgcctcag atattaatta atagagaacc tttgcctcat ctgcattttg atgtagagct 2341 tcttggagac tgtgatgtca taattaatga attgtgtcat aggttaggtg gtgaatatgc 2401 caaactttgc tgtaaccctg taaagctttc agaaattact gaaaaacctc cacgaacaca 2461 aaaagaattg gcttatttgt cagagttgcc acccacacct cttcatgttt cagaagactc 2521 aagttcacca gaaagaactt caccaccaga ttcttcagtg attgtcacac ttttagacca 2581 agcagctaag agtaatgatg atttagatgt gtctgaatca aaaggttgta tggaagaaaa 2641 accacaggaa gtacaaactt ctaggaatgt tgaaagtatt gctgaacaga tggaaaatcc 2701 ggatttgaag aatgttggtt ctagtactgg ggagaaaaat gaaagaactt cagtggctgg 2761 aacagtgaga aaatgctggc ctaatagagt ggcaaaggag cagattagta ggcggcttga 2821 tggtaatcag tatctgtttt tgccaccaaa tcgttacatt ttccatggcg ctgaggtata 2881 ttcagactct gaagatgacg tcttatcctc tagttcttgt ggcagtaaca gtgatagtgg 2941 gacatgccag agtccaagtt tagaagaacc catggaggat gaaagtgaaa ttgaagaatt 3001 ctacaatggc ttagaagatg agcctgatgt tccagagaga gctggaggag ctggatttgg 3061 gactgatgga gatgatcaag aggcaattaa tgaagctata tctgtgaaac aggaagtaac 3121 agacatgaac tatccatcaa acaaatcata gctcgagtct agagggcccg tttaaacccg 3181 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt 3241 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat 3301 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag 3361 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc 3421 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg 3481 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc 3541 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc 3601 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct 3661 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac 3721 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac 3781 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat 3841 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg 3901 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg 3961 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca 4021 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact 4081 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta 4141 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag 4201 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc 4261 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga 4321 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa 4381 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt 4441 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg 4501 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa 4561 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac 4621 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt 4681 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact 4741 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg 4801 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg 4861 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc 4921 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt 4981 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc 5041 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg 5101 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg 5161 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct 5221 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta 5281 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat 5341 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct 5401 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt 5461 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag 5521 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt 5581 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag 5641 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg 5701 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat 5761 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta 5821 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa 5881 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc 5941 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt 6001 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca 6061 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg 6121 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat 6181 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta 6241 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct 6301 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac 6361 aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag 6421 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact 6481 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa 6541 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt 6601 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag 6661 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca 6721 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc 6781 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt 6841 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg 6901 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca 6961 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg 7021 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca 7081 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg 7141 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct 7201 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca 7261 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca 7321 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg 7381 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac 7441 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt 7501 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc 7561 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc 7621 cctatggt //