LOCUS       sequence-152138-        7567 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 7567)
  AUTHORS   Vaziri H, Dessain SK, Ng Eaton E, Imai SI, Frye RA, Pandita TK, 
            Guarente L, Weinberg RA
  TITLE     hSIR2(SIRT1) functions as an NAD-dependent p53 deacetylase.
  JOURNAL   Cell 2001 Oct 19;107(2):149-59.
  PUBMED    11672523
REFERENCE   2  (bases 1 to 7567)
  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: "Cell"; 
            date: "2001-10-19"; volume: "107(2)"; pages: "149-59"
COMMENT     Sequence Label: pYESir2HY-puro
FEATURES             Location/Qualifiers
     source          1..7567
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             134..550
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     480..502
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     518..534
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             564..2807
                     /codon_start=1
                     /product="human NAD+-dependent protein deacetylase 
                     sirtuin-1"
                     /label=SIRT1
                     /note="homolog of budding yeast Sir2"
                     /translation="MADEAALALQPGGSPSAAGADREAASSPAGEPLRKRPRRDGPGLE
                     RSPGEPGGAAPEREVPAAARGCPGAAAAALWREAEAEAAAAGGEQEAQATAAAGEGDNG
                     PGLQGPSREPPLADNLYDEDDDDEGEEEEEAAAAAIGYRDNLLFGDEIITNGFHSCESD
                     EEDRASHASSSDWTPRPRIGPYTFVQQHLMIGTDPRTILKDLLPETIPPPELDDMTLWQ
                     IVINILSEPPKRKKRKDINTIEDAVKLLQECKKIIVLTGAGVSVSCGIPDFRSRDGIYA
                     RLAVDFPDLPDPQAMFDIEYFRKDPRPFFKFAKEIYPGQFQPSLCHKFIALSDKEGKLL
                     RNYTQNIDTLEQVAGIQRIIQCYGSFATASCLICKYKVDCEAVRGDIFNQVVPRCPRCP
                     ADEPLAIMKPEIVFFGENLPEQFHRAMKYDKDEVDLLIVIGSSLKVRPVALIPSSIPHE
                     VPQILINREPLPHLHFDVELLGDCDVIINELCHRLGGEYAKLCCNPVKLSEITEKPPRT
                     QKELAYLSELPPTPLHVSEDSSSPERTSPPDSSVIVTLLDQAAKSNDDLDVSESKGCME
                     EKPQEVQTSRNVESIAEQMENPDLKNVGSSTGEKNERTSVAGTVRKCWPNRVAKEQISR
                     RLDGNQYLFLPPNRYIFHGAEVYSDSEDDVLSSSSCGSNSDSGTCQSPSLEEPMEDESE
                     IEEFYNGLEDEPDVPERAGGAGFGTDGDDQEAINEAISVKQEVTDMNYPSNKS"
     primer_bind     complement(2882..2901)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2883..2901)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2908..2925)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2908..2924)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2917..2939)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3089..3109)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3094..3423
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3274..3409
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3433..4032
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3433..3452)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3929..3949
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4253..4722
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4852..4869)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5023..5611)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5103..5122)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5771..6562,6563..6631))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6394..6413
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6632..6736)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             6767..7236
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    7301..7500
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
ORIGIN
        1 acagttcccg cctccgtctg aatttttgct ttcggtttgg aaccgaagcc gcgcgtcttg
       61 tctgctgcag cgctgcagca tcgttctgtg ttgtctctgt ctgactgtgt ttctgtattt
      121 gtctgaaaat tagggccaga ctgttaccac tcccttaagt ttgaccttag gtcactggaa
      181 agatgtcgag cggatcgctc acaaccagtc ggtagatgtc aagaagagac gttgggttac
      241 cttctgctct gcagaatggc caacctttaa cgtcggatgg ccgcgagacg gcacctttaa
      301 ccgagacctc atcacccagg ttaagatcaa ggtcttttca cctggcccgc atggacaccc
      361 agaccaggtc ccctacatcg tgacctggga agccttggct tttgaccccc ctccctgggt
      421 caagcccttt gtacacccta agcctccgcc tcctcttcct ccatccgccc cgtctctccc
      481 ccttgaacct cctcgttcga ccccgcctcg atcctccctt tatccagccc tcactccttc
      541 tctaggcgcc ggccggatcc accatggcgg acgaggcggc cctcgccctt cagcccggcg
      601 gctccccctc ggcggcgggg gccgacaggg aggccgcgtc gtcccccgcc ggggagccgc
      661 tccgcaagag gccgcggaga gatggtcccg gcctcgagcg gagcccgggc gagcccggtg
      721 gggcggcccc agagcgtgag gtgccggcgg cggccagggg ctgcccgggt gcggcggcgg
      781 cggcgctgtg gcgggaggcg gaggcagagg cggcggcggc aggcggggag caagaggccc
      841 aggcgactgc ggcggctggg gaaggagaca atgggccggg cctgcagggc ccatctcggg
      901 agccaccgct ggccgacaac ttgtacgacg aagacgacga cgacgagggc gaggaggagg
      961 aagaggcggc ggcggcggcg attgggtacc gagataacct tctgttcggt gatgaaatta
     1021 tcactaatgg ttttcattcc tgtgaaagtg atgaggagga tagagcctca catgcaagct
     1081 ctagtgactg gactccaagg ccacggatag gtccatatac ttttgttcag caacatctta
     1141 tgattggcac agatcctcga acaattctta aagatttatt gccggaaaca atacctccac
     1201 ctgagttgga tgatatgaca ctgtggcaga ttgttattaa tatcctttca gaaccaccaa
     1261 aaaggaaaaa aagaaaagat attaatacaa ttgaagatgc tgtgaaatta ctgcaagagt
     1321 gcaaaaaaat tatagttcta actggagctg gggtgtctgt ttcatgtgga atacctgact
     1381 tcaggtcaag ggatggtatt tatgctcgcc ttgctgtaga cttcccagat cttccagatc
     1441 ctcaagcgat gtttgatatt gaatatttca gaaaagatcc aagaccattc ttcaagtttg
     1501 caaaggaaat atatcctgga caattccagc catctctctg tcacaaattc atagccttgt
     1561 cagataagga aggaaaacta cttcgcaact atacccagaa catagacacg ctggaacagg
     1621 ttgcgggaat ccaaaggata attcagtgtt atggttcctt tgcaacagca tcttgcctga
     1681 tttgtaaata caaagttgac tgtgaagctg tacgaggaga tatttttaat caggtagttc
     1741 ctcgatgtcc taggtgccca gctgatgaac cgcttgctat catgaaacca gagattgtgt
     1801 tttttggtga aaatttacca gaacagtttc atagagccat gaagtatgac aaagatgaag
     1861 ttgacctcct cattgttatt gggtcttccc tcaaagtaag accagtagca ctaattccaa
     1921 gttccatacc ccatgaagtg cctcagatat taattaatag agaacctttg cctcatctgc
     1981 attttgatgt agagcttctt ggagactgtg atgtcataat taatgaattg tgtcataggt
     2041 taggtggtga atatgccaaa ctttgctgta accctgtaaa gctttcagaa attactgaaa
     2101 aacctccacg aacacaaaaa gaattggctt atttgtcaga gttgccaccc acacctcttc
     2161 atgtttcaga agactcaagt tcaccagaaa gaacttcacc accagattct tcagtgattg
     2221 tcacactttt agaccaagca gctaagagta atgatgattt agatgtgtct gaatcaaaag
     2281 gttgtatgga agaaaaacca caggaagtac aaacttctag gaatgttgaa agtattgctg
     2341 aacagatgga aaatccggat ttgaagaatg ttggttctag tactggggag aaaaatgaaa
     2401 gaacttcagt ggctggaaca gtgagaaaat gctggcctaa tagagtggca aaggagcaga
     2461 ttagtaggcg gcttgatggt aatcagtatc tgtttttgcc accaaatcgt tacattttcc
     2521 atggcgctga ggtatattca gactctgaag atgacgtctt atcctctagt tcttgtggca
     2581 gtaacagtga tagtgggaca tgccagagtc caagtttaga agaacccatg gaggatgaaa
     2641 gtgaaattga agaattctac aatggcttag aagatgagcc tgatgttcca gagagagctg
     2701 gaggagctgg atttgggact gatggagatg atcaagaggc aattaatgaa gctatatctg
     2761 tgaaacagga agtaacagac atgaactatc catcaaacaa atcatagtgt aataattgtg
     2821 caggtacagg aattgttcca ctctagacaa gggcgaattc gcggccgcta aattcaattc
     2881 gccctatagt gagtcgtatt acaattcact ggccgtcgtt ttacaacgtc gtgactggga
     2941 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     3001 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tatacgtagg
     3061 aattcgccag cacagtggtc gaccctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     3121 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     3181 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3241 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     3301 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     3361 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     3421 aaaaagctta ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
     3481 cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
     3541 gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
     3601 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
     3661 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
     3721 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac
     3781 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc
     3841 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg
     3901 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
     3961 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag
     4021 cccggtgcct gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc
     4081 catgcatcga taaaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac
     4141 cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc atggaaaaat
     4201 acataactga gaatagagaa gttcagatca aggtcaggaa cagatggaac agctgaatat
     4261 gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat
     4321 ggaacagctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
     4381 gggccaagaa cagatggtcc ccagatgcgg tccagccctc agcagtttct agagaaccat
     4441 cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc
     4501 aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc
     4561 ccacaacccc tcactcgggg cgccagtcct ccgattgact gagtcgcccg ggtacccgtg
     4621 tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc cttgggaggg
     4681 tctcctctga gtgattgact acccgtcagc gggggtcttt cacatgcagc atgtatcaaa
     4741 attaatttgg ttttttttct taagtattta cattaaatgg ccatagttgc attaatgaat
     4801 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     4861 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     4921 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     4981 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5041 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5101 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5161 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     5221 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5281 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5341 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5401 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5461 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5521 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5581 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5641 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5701 gatcttcacc tagatccttt tgcggccgca aatcaatcta aagtatatat gagtaaactt
     5761 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     5821 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     5881 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     5941 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6001 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6061 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     6121 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     6181 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     6241 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     6301 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     6361 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     6421 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     6481 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     6541 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     6601 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     6661 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     6721 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgcagcctga atatgggcca
     6781 aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggaaca
     6841 gctgaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca
     6901 agaacagatg gtccccagat gcggtccagc cctcagcagt ttctagagaa ccatcagatg
     6961 tttccagggt gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag
     7021 ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa
     7081 cccctcactc ggggcgccag tcctccgatt gactgagtcg cccgggtacc cgtgtatcca
     7141 ataaaccctc ttgcagttgc atccgacttg tggtctcgct gttccttggg agggtctcct
     7201 ctgagtgatt gactacccgt cagcgggggt ctttcatttg ggggctcgtc cgggatcggg
     7261 agacccctgc ccagggacca ccgacccacc accgggaggc aagctggcca gcaacttatc
     7321 tgtgtctgtc cgattgtcta gtgtctatga ctgattttat gcgcctgcgt cggtactagt
     7381 tagctaacta gctctgtatc tggcggaccc gtggtggaac tgacgagttc tgaacacccg
     7441 gccgcaaccc tgggagacgt cccagggact ttgggggccg tttttgtggc ccgacctgag
     7501 gaagggagtc gatgtggaat ccgaccccgt caggatatgt ggttctggta ggagacgaga
     7561 acctaaa
//
