LOCUS       V031235                 7058 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031235
VERSION     V031235
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     primer_bind     166..182
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     enhancer        343..722
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        723..926
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        952..970
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             1010..3220
                     /codon_start=1
                     /gene=""
                     /note="homolog of budding yeast Sir2"
                     /product="mouse NAD+-dependent protein deacetylase
                     sirtuin-1"
                     /protein_id=""
                     /transl_table=1
                     /translation="MADEVALALQAAGSPSAAAAMEAASQPADEPLRKRPRRDGPGLGR
                     SPGEPSAAVAPAAAGCEAASAAAPAALWREAAGAAASAEREAPATAVAGDGDNGSGLRR
                     EPRAADDFDDDEGEEEDEAAAAAAAAAIGYRDNLLLTDGLLTNGFHSCESDDDDRTSHA
                     SSSDWTPRPRIGPYTFVQQHLMIGTDPRTILKDLLPETIPPPELDDMTLWQIVINILSE
                     PPKRKKRKDINTIEDAVKLLQECKKIIVLTGAGVSVSCGIPDFRSRDGIYARLAVDFPD
                     LPDPQAMFDIEYFRKDPRPFFKFAKEIYPGQFQPSLCHKFIALSDKEGKLLRNYTQNID
                     TLEQVAGIQRILQCHGSFATASCLICKYKVDCEAVRGDIFNQVVPRCPRCPADEPLAIM
                     KPEIVFFGENLPEQFHRAMKYDKDEVDLLIVIGSSLKVRPVALIPSSIPHEVPQILINR
                     EPLPHLHFDVELLGDCDVIINELCHRLGGEYAKLCCNPVKLSEITEKPPRPQKELVHLS
                     ELPPTPLHISEDSSSPERTVPQDSSVIATLVDQATNNNVNDLEVSESSCVEEKPQEVQT
                     SRNVENINVENPDFKAVGSSTADKNERTSVAETVRKCWPNRLAKEQISKRLEGNQYLFV
                     PPNRYIFHGAEVYSDSEDDVLSSSSCGSNSDSGTCQSPSLEEPLEDESEIEEFYNGLED
                     DTERPECAGGSGFGADGGDQEVVNEAIATRQELTDVNYPSDKS"
                     /label="SIRT1"
     CDS             3224..3253
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             3272..3295
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(3315..3331)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    3342..3964
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(4113..4701)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    5030..5077
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(5309..6103)
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     promoter        complement(6138..6495)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(6497..6601)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      6628..7056
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
ORIGIN
        1 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag
       61 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
      121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
      181 gtgccaagct gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt
      241 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata
      301 tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta
      361 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      421 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      481 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      541 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      601 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      661 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      721 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat
      781 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      841 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac
      901 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagaatttt gtaatacgac
      961 tcactatagg gcggccggga attcgtcgac tggatccggt accgccacca tggcggacga
     1021 ggtggcgctc gcccttcagg ccgccggctc cccttccgcg gcggccgcca tggaggccgc
     1081 gtcgcagccg gcggacgagc cgctccgcaa gaggccccgc cgagacgggc ctggcctcgg
     1141 gcgcagcccg ggcgagccga gcgcagcagt ggcgccggcg gccgcggggt gtgaggcggc
     1201 gagcgccgcg gccccggcgg cgctgtggcg ggaggcggca ggggcggcgg cgagcgcgga
     1261 gcgggaggcc ccggcgacgg ccgtggccgg ggacggagac aatgggtccg gcctgcggcg
     1321 ggagccgagg gcggctgacg acttcgacga cgacgagggc gaggaggagg acgaggcggc
     1381 ggcggcagcg gcggcggcag cgatcggcta ccgagacaac ctcctgttga ccgatggact
     1441 cctcactaat ggctttcatt cctgtgaaag tgatgacgat gacagaacgt cacacgccag
     1501 ctctagtgac tggactccgc ggccgcggat aggtccatat acttttgttc agcaacatct
     1561 catgattggc accgatcctc gaacaattct taaagattta ttaccagaaa caattcctcc
     1621 acctgagctg gatgatatga cgctgtggca gattgttatt aatatccttt cagaaccacc
     1681 aaagcggaaa aaaagaaaag atatcaatac aattgaagat gctgtgaagt tactgcagga
     1741 gtgtaaaaag ataatagttc tgactggagc tggggtttct gtctcctgtg ggattcctga
     1801 cttcagatca agagacggta tctatgctcg ccttgcggtg gacttcccag acctcccaga
     1861 ccctcaagcc atgtttgata ttgagtattt tagaaaagac ccaagaccat tcttcaagtt
     1921 tgcaaaggaa atatatcccg gacagttcca gccgtctctg tgtcacaaat tcatagcttt
     1981 gtcagataag gaaggaaaac tacttcgaaa ttatactcaa aatatagata ccttggagca
     2041 ggttgcagga atccaaagga tccttcagtg tcatggttcc tttgcaacag catcttgcct
     2101 gatttgtaaa tacaaagttg attgtgaagc tgttcgtgga gacattttta atcaggtagt
     2161 tcctcggtgc cctaggtgcc cagctgatga gccacttgcc atcatgaagc cagagattgt
     2221 cttctttggt gaaaacttac cagaacagtt tcatagagcc atgaagtatg acaaagatga
     2281 agttgacctc ctcattgtta ttggatcttc tctgaaagtg agaccagtag cactaattcc
     2341 aagttctata ccccatgaag tgcctcaaat attaataaat agggaacctt tgcctcatct
     2401 acattttgat gtagagctcc ttggagactg cgatgttata attaatgagt tgtgtcatag
     2461 gctaggtggt gaatatgcca aactttgttg taaccctgta aagctttcag aaattactga
     2521 aaaacctcca cgcccacaaa aggaattggt tcatttatca gagttgccac caacacctct
     2581 tcatatttcg gaagactcaa gttcacctga aagaactgta ccacaagact cttctgtgat
     2641 tgctacactt gtagaccaag caacaaacaa caatgttaat gatttagaag tatctgaatc
     2701 aagttgtgtg gaagaaaaac cacaagaagt acagactagt aggaatgttg agaacattaa
     2761 tgtggaaaat ccagatttta aggctgttgg ttccagtact gcagacaaaa atgaaagaac
     2821 ttcagttgca gaaacagtga gaaaatgctg gcctaataga cttgcaaagg agcagattag
     2881 taagcggctt gagggtaatc aatacctgtt tgtaccacca aatcgttaca tattccacgg
     2941 tgctgaggta tactcagact ctgaagatga cgtcttgtcc tctagttcct gtggcagtaa
     3001 cagtgacagt ggcacatgcc agagtccaag tttagaagaa cccttggaag atgaaagtga
     3061 aattgaagaa ttctacaatg gcttggaaga tgatacggag aggcccgaat gtgctggagg
     3121 atctggattt ggagctgatg gaggggatca agaggttgtt aatgaagcta tagctacaag
     3181 acaggaattg acagatgtaa actatccatc agacaaatca ctcgagcaga aactcatctc
     3241 agaagaggat ctggcagcaa atgatatcct ggattacaag gatgacgacg ataaggttta
     3301 aacggccggc cgcggtcata gctgtttcct gaacagatcc cgggtggcat ccctgtgacc
     3361 cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc agccttgtcc
     3421 taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat attatggggt
     3481 ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg gcctgcgggg
     3541 tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca atctccgcct
     3601 cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc caggcatgca
     3661 tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc atattggcca
     3721 ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc aaattgctgg
     3781 gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttaaaa taactatacc
     3841 agcaggagga cgtccagaca cagcataggc tacctggcca tgcccaaccg gtgggacatt
     3901 tgagttgctt gcttggcact gtcctctcat gcgttgggtc cactcagtag atgcctgttg
     3961 aattgggtac gcggccagcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4021 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4081 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4141 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4201 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4261 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4321 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4381 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4441 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4501 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     4561 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4621 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4681 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4741 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     4801 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaacct
     4861 gaggctatgg cagggcctgc cgccccgacg ttggctgcga gccctgggcc ttcacccgaa
     4921 cttggggggt ggggtgggga aaaggaagaa acgcgggcgt attggcccca atggggtctc
     4981 ggtggggtat cgacagagtg ccagccctgg gaccgaaccc cgcgtttatg aacaaacgac
     5041 ccaacaccgt gcgttttatt ctgtcttttt attgccgtca tagcgcgggt tccttccggt
     5101 attgtctcct tccgtgtttc agttagcctc cccctagggt gggcgaagaa ctccagcatg
     5161 agatccccgc gctggaggat catccagccg gcgtcccgga aaacgattcc gaagcccaac
     5221 ctttcataga aggcggcggt ggaatcgaaa tctcgtgatg gcaggttggg cgtcgcttgg
     5281 tcggtcattt cgaaccccag agtcccgctc agaagaactc gtcaagaagg cgatagaagg
     5341 cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt
     5401 cgccgccaag ctcttcagca atatcacggg tagccaacgc tatgtcctga tagcgatccg
     5461 ccacacccag ccggccacag tcgatgaatc cagaaaagcg gccattttcc accatgatat
     5521 tcggcaagca ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc atgctcgcct
     5581 tgagcctggc gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc agatcatcct
     5641 gatcgacaag accggcttcc atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt
     5701 ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca tcagccatga
     5761 tggatacttt ctcggcagga gcaaggtgag atgacaggag atcctgcccc ggcacttcgc
     5821 ccaatagcag ccagtccctt cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa
     5881 cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc ttgcagttca ttcagggcac
     5941 cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg
     6001 cggcatcaga gcagccgatt gtctgttgtg cccagtcata gccgaatagc ctctccaccc
     6061 aagcggccgg agaacctgcg tgcaatccat cttgttcaat catgcgaaac gatcctcatc
     6121 ctgtctcttg atcgatcttt gcaaaagcct aggcctccaa aaaagcctcc tcactacttc
     6181 tggaatagct cagaggccga ggcggcctcg gcctctgcat aaataaaaaa aattagtcag
     6241 ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg
     6301 ggcgggacta tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg
     6361 gagcctgggg actttccaca cctggttgct gactaattga gatgcatgct ttgcatactt
     6421 ctgcctgctg gggagcctgg ggactttcca caccctaact gacacacatt ccacagctgg
     6481 ttctttccgc ctcaggactc ttcctttttc aatattattg aagcatttat cagggttatt
     6541 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     6601 gcacatttcc ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta agcgcggcgg
     6661 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     6721 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     6781 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     6841 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     6901 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     6961 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     7021 aaaatgagct gatttaacaa aaatttaacg cgaatttt
//
