LOCUS       sequence-43243-e        5420 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5420)
  AUTHORS   Brunet A, Sweeney LB, Sturgill JF, Chua KF, Greer PL, Lin Y, Tran H,
            Ross SE, Mostoslavsky R, Cohen HY, Hu LS, Cheng HL, Jedrychowski MP,
            Gygi SP, Sinclair DA, Alt FW, Greenberg ME
  TITLE     Stress-dependent regulation of FOXO transcription factors by the 
            SIRT1 deacetylase.
  JOURNAL   Science 2004 Mar 26;303(5666):2011-5. Epub 2004 Feb 19.
  PUBMED    14976264
REFERENCE   2  (bases 1 to 5420)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science"; 
            date: "2004-03-26"; volume: "303(5666)"; pages: "2011-5. Epub 2004 
            Feb 19"
COMMENT     Sequence Label: Flag-SIRT1 H363Y
FEATURES             Location/Qualifiers
     source          1..5420
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1420..1443)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     1772..1792
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1847..1951
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1952..2020,2021..2812)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2170..2189)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2983..3571
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3472..3491
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3725..3742
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4007..4026
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     polyA_signal    4085..4219
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4109..4128)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4163..4182
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4455..4477
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4469..4486
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4470..4486
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4493..4512
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4493..4511
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(join(4587..5420,1..1407))
                     /codon_start=1
                     /product="human NAD+-dependent protein deacetylase 
                     sirtuin-1"
                     /label=SIRT1
                     /note="homolog of budding yeast Sir2"
                     /translation="ADEAALALQPGGSPSAAGADREAASSPAGEPLRKRPRRDGPGLER
                     SPGEPGGAAPEREVPAAARGCPGAAAAALWREAEAEAAAAGGEQEAQATAAAGEGDNGP
                     GLQGPSREPPLADNLYDEDDDDEGEEEEEAAAAAIGYRDNLLFGDEIITNGFHSCESDE
                     EDRASHASSSDWTPRPRIGPYTFVQQHLMIGTDPRTILKDLLPETIPPPELDDMTLWQI
                     VINILSEPPKRKKRKDINTIEDAVKLLQECKKIIVLTGAGVSVSCGIPDFRSRDGIYAR
                     LAVDFPDLPDPQAMFDIEYFRKDPRPFFKFAKEIYPGQFQPSLCHKFIALSDKEGKLLR
                     NYTQNIDTLEQVAGIQRIIQCYGSFATASCLICKYKVDCEAVRGDIFNQVVPRCPRCPA
                     DEPLAIMKPEIVFFGENLPEQFHRAMKYDKDEVDLLIVIGSSLKVRPVALIPSSIPHEV
                     PQILINREPLPHLHFDVELLGDCDVIINELCHRLGGEYAKLCCNPVKLSEITEKPPRTQ
                     KELAYLSELPPTPLHVSEDSSSPERTSPPDSSVIVTLLDQAAKSNDDLDVSESKGCMEE
                     KPQEVQTSRNVESIAEQMENPDLKNVGSSTGEKNERTSVAGTVRKCWPNRVAKEQISRR
                     LDGNQYLFLPPNRYIFHGAEVYSDSEDDVLSSSSCGSNSDSGTCQSPSLEEPMEDESEI
                     EEFYNGLEDEPDVPERAGGAGFGTDGDDQEAINEAISVKQEVTDMNYPSNKS"
ORIGIN
        1 aggcaaaggt tctctattaa ttaatatctg aggcacttca tggggtatgg aacttggaat
       61 tagtgctact ggtcttactt tgagggaaga cccaataaca atgaggaggt caacttcatc
      121 tttgtcatac ttcatggctc tatgaaactg ttctggtaaa ttttcaccaa aaaacacaat
      181 ctctggtttc atgatagcaa gcggttcatc agctgggcac ctaggacatc gaggaactac
      241 ctgattaaaa atatctcctc gtacagcttc acagtcaact ttgtatttac aaatcaggca
      301 agatgctgtt gcaaaggaac cataacactg aattatcctt tggattcccg caacctgttc
      361 cagcgtgtct atgttctggg tatagttgcg aagtagtttt ccttccttat ctgacaaggc
      421 tatgaatttg tgacagagag atggctggaa ttgtccagga tatatttcct ttgcaaactt
      481 gaagaatggt cttggatctt ttctgaaata ttcaatatca aacatcgctt gaggatctgg
      541 aagatctggg aagtctacag caaggcgagc ataaatacca tcccttgacc tgaagtcagg
      601 tattccacat gaaacagaca ccccagctcc agttagaact ataatttttt tgcactcttg
      661 cagtaatttc acagcatctt caattgtatt aatatctttt ctttttttcc tttttggtgg
      721 ttctgaaagg atattaataa caatctgcca cagtgtcata tcatccaact caggtggagg
      781 tattgtttcc ggcaataaat ctttaagaat tgttcgagga tctgtgccaa tcataagatg
      841 ttgctgaaca aaagtatatg gacctatccg tggccttgga gtccagtcac tagagcttgc
      901 atgtgaggct ctatcctcct catcactttc acaggaatga aaaccattag tgataatttc
      961 atcaccgaac agaaggttat ctcggtaccc aatcgccgcc gccgccgcct cttcctcctc
     1021 ctcgccctcg tcgtcgtcgt cttcgtcgta caagttgtcg gccagcggtg gctcccgaga
     1081 tgggccctgc aggcccggcc cattgtctcc ttccccagcc gccgcagtcg cctgggcctc
     1141 ttgctccccg cctgccgccg ccgcctctgc ctccgcctcc cgccacagcg ccgccgccgc
     1201 cgcacccggg cagcccctgg ccgccgccgg cacctcacgc tctggggccg ccccaccggg
     1261 ctcgcccggg ctccgctcga ggccgggacc atctctccgc ggcctcttgc ggagcggctc
     1321 cccggcgggg gacgacgcgg cctccctgtc ggcccccgcc gccgaggggg agccgccggg
     1381 ctgaagggcg agggccgcct cgtccgccga attcccgggt ttatcgtcat cgtccttgta
     1441 gtccatcctg aggatccgag ctcggtacca agcttagatc tcctccaaaa aagcctcttc
     1501 actacttctg gaatagctca gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa
     1561 ttagtcagcc atggggcgga gaatgggcgg aactgggcgg agttaggggc gggatgggcg
     1621 gagttagggg cgggactatg gttgctgact aattgagatg catgctttgc atacttctgc
     1681 ctgctgggga gcctggggac tttccttgct gactaattga gatgcatgct ttgcatactt
     1741 ctgcctgctg gggagcctgg ggactttcca caccctaact gacacacatt ccacagctgg
     1801 ttctttccgc ctcagaaggt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     1861 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     1921 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     1981 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     2041 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     2101 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     2161 taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg
     2221 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     2281 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     2341 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     2401 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     2461 cataccaaac gacgagcgtg acaccacgat gcctgcagca atggcaacaa cgttgcgcaa
     2521 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     2581 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     2641 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     2701 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     2761 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     2821 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     2881 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     2941 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     3001 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     3061 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     3121 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3181 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3241 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3301 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3361 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3421 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     3481 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     3541 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     3601 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     3661 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     3721 gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac
     3781 gcatctgtgc ggtatttcac accgcatatc gacgctctcc cttatgcgac tcctgcatta
     3841 ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat
     3901 gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga
     3961 aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga
     4021 tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt
     4081 agaggatcca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     4141 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     4201 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     4261 ggaggtgtgg gaggtttttt aaagcaagta aaacctctac agatgtgata tggctgatta
     4321 tgatcattac ttatctaggt ataggctgcg caactgttgg gaagggcgat cggtgcgggc
     4381 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     4441 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat tgtaatacga
     4501 ctcactatag ggcgaattga atttagcggc cgcgaattcg cccttgtcta gagtggaaca
     4561 attcctgtac ctgcacaatt attacactat gatttgtttg atggatagtt catgtctgtt
     4621 acttcctgtt tcacagatat agcttcatta attgcctctt gatcatctcc atcagtccca
     4681 aatccagctc ctccagctct ctctggaaca tcaggctcat cttctaagcc attgtagaat
     4741 tcttcaattt cactttcatc ctccatgggt tcttctaaac ttggactctg gcatgtccca
     4801 ctatcactgt tactgccaca agaactagag gataagacgt catcttcaga gtctgaatat
     4861 acctcagcgc catggaaaat gtaacgattt ggtggcaaaa acagatactg attaccatca
     4921 agccgcctac taatctgctc ctttgccact ctattaggcc agcattttct cactgttcca
     4981 gccactgaag ttctttcatt tttctcccca gtactagaac caacattctt caaatccgga
     5041 ttttccatct gttcagcaat actttcaaca ttcctagaag tttgtacttc ctgtggtttt
     5101 tcttccatac aaccttttga ttcagacaca tctaaatcat cattactctt agctgcttgg
     5161 tctaaaagtg tgacaatcac tgaagaatct ggtggtgaag ttctttctgg tgaacttgag
     5221 tcttctgaaa catgaagagg tgtgggtggc aactctgaca aataagccaa ttctttttgt
     5281 gttcgtggag gtttttcagt aatttctgaa agctttacag ggttacagca aagtttggca
     5341 tattcaccac ctaacctatg acacaattca ttaattatga catcacagtc tccaagaagc
     5401 tctacatcaa aatgcagatg
//
