LOCUS V033915 6185 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V033915 VERSION V033915 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..1678 /label="Sirt3(NM_022433)" /note="Sirt3(NM_022433)" /gene="Sirt3" CDS 1685..1714 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /transl_table=1 /translation="EQKLISEEDL" /label="Myc" polyA_signal 1752..1976 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2022..2450 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2464..2793 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2860..3654 /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 3828..3949 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(3998..4014) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4022..4038 /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(4046..4076) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4091..4112 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4400..4985) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5156..6016) /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(6017..6121) /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 gtgggggccg gcatcagcac acccagtggc atcccggact tcagatcccc 961 agggagcggc ctctacagca accttcagca gtatgacatc ccgtaccctg aagccatctt 1021 tgaacttggc tttttctttc acaaccccaa gccctttttc atgttggcca aggagctgta 1081 ccctgggcac tacaggccca atgtcactca ctacttcctg aggctcctcc acgacaagga 1141 gctgcttctg cggctctata cacagaacat cgacgggctt gagagagcat ctgggatccc 1201 tgcctcaaag ctggttgaag cccacgggac ctttgtaaca gctacatgca cggtctgtcg 1261 aaggtccttc ccaggggaag acatatgggc tgatgtgatg gcggacaggg tgccccgctg 1321 ccctgtctgt actggcgttg tgaaacccga cattgtgttc tttggggagc agctgcctgc 1381 aaggttccta ctccatatgg ctgacttcgc tttggcagat ctgctactca ttcttgggac 1441 ctccctggag gtggagcctt ttgccagctt gtctgaagca gtacagaaat cagtgccccg 1501 actgctcatc aatcgagact tggtggggcc gttcgttctg agtcctcgaa ggaaagatgt 1561 ggtccagcta ggggatgtag ttcatggtgt ggaaaggctg gtggacctcc tggggtggac 1621 acaagaactg ctggatctta tgcagcggga acgtggcaag ctggatggac aggacagact 1681 cgaggagcag aagctgatct cagaggagga cctgtagtga gggcccgttt aaacccgctg 1741 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc 1801 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc 1861 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa 1921 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc 1981 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc 2041 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct 2101 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg 2161 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga 2221 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt 2281 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg 2341 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc 2401 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg 2461 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa 2521 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc 2581 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg 2641 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt 2701 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga 2761 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt 2821 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg 2881 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag 2941 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt 3001 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta 3061 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg 3121 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt 3181 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat 3241 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg 3301 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca 3361 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc 3421 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc 3481 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat 3541 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc 3601 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga 3661 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt 3721 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga 3781 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg 3841 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt 3901 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta 3961 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa 4021 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct 4081 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc 4141 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg 4201 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc 4261 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag 4321 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa 4381 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc 4441 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc 4501 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg 4561 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt 4621 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc 4681 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc 4741 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag 4801 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg 4861 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa 4921 ccaccgctgg tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat 4981 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac 5041 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt 5101 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc 5161 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg 5221 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg 5281 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc 5341 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta 5401 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg 5461 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct 5521 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta 5581 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg 5641 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga 5701 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt 5761 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca 5821 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt 5881 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt 5941 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga 6001 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt 6061 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc 6121 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct 6181 atggt //