LOCUS       sequence-207904-        5599 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression construct of N terminus of SIRT1: Exon1-Exon3 
            (deltaExon2).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5599)
  AUTHORS   Deota S, Chattopadhyay T, Ramachandran D, Armstrong E, Camacho B, 
            Maniyadath B, Fulzele A, Gonzalez-de-Peredo A, Denu JM, 
            Kolthur-Seetharam U
  TITLE     Identification of a Tissue-Restricted Isoform of SIRT1 Defines a 
            Regulatory Domain that Encodes Specificity.
  JOURNAL   Cell Rep. 2017 Mar 28;18(13):3069-3077. doi: 
            10.1016/j.celrep.2017.03.012.
  PUBMED    28355560
REFERENCE   2  (bases 1 to 5599)
  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"; doi: 
            "10.1016/j.celrep.2017.03"; journalName: "Cell Rep"; date: 
            "2017-03-28- 28"; volume: "18"; issue: "13"; pages: "3069-3077"
COMMENT     Sequence Label: pGEX-4T-3 SIRT1-NTerm Exon1-Exon3
FEATURES             Location/Qualifiers
     source          1..5599
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..22,23..29)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    37..53
                     /label=lac operator
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     CDS             76..729
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     687..709
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             736..753
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     regulatory      808..817
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1467..1489)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1634..1652)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1720..1824
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1825..1893,1894..2685)
                     /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(2043..2062)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2856..3444
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3345..3364
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3598..3615
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3688..3765
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3766..4848
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(3785..3804)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(4897..4902,4903..4920,4921..4927)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4935..4951
                     /label=lac operator
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     4959..4975
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4959..4975
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4971..5498
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(4991..5008)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4991..5007)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5000..5022)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca
       61 caggaaacag tattcatgtc ccctatacta ggttattgga aaattaaggg ccttgtgcaa
      121 cccactcgac ttcttttgga atatcttgaa gaaaaatatg aagagcattt gtatgagcgc
      181 gatgaaggtg ataaatggcg aaacaaaaag tttgaattgg gtttggagtt tcccaatctt
      241 ccttattata ttgatggtga tgttaaatta acacagtcta tggccatcat acgttatata
      301 gctgacaagc acaacatgtt gggtggttgt ccaaaagagc gtgcagagat ttcaatgctt
      361 gaaggagcgg ttttggatat tagatacggt gtttcgagaa ttgcatatag taaagacttt
      421 gaaactctca aagttgattt tcttagcaag ctacctgaaa tgctgaaaat gttcgaagat
      481 cgtttatgtc ataaaacata tttaaatggt gatcatgtaa cccatcctga cttcatgttg
      541 tatgacgctc ttgatgttgt tttatacatg gacccaatgt gcctggatgc gttcccaaaa
      601 ttagtttgtt ttaaaaaacg tattgaagct atcccacaaa ttgataagta cttgaaatcc
      661 agcaagtata tagcatggcc tttgcagggc tggcaagcca cgtttggtgg tggcgaccat
      721 cctccaaaat cggatctggt tccgcgtgga tccatggcgg acgaggtggc gctcgccctt
      781 caggccgccg gctccccttc cgcggcggcc gccatggagg ccgcgtcgca gccggcggac
      841 gagccgctcc gcaagaggcc ccgccgagac gggcctggcc tcgggcgcag cccgggcgag
      901 ccgagcgcag cagtggcgcc ggcggccgcg gggtgtgagg cggcgagcgc cgcggccccg
      961 gcggcgctgt ggcgggaggc ggcaggggcg gcggcgagcg cggagcggga ggccccggcg
     1021 acggccgtgg ccggggacgg agacaatggg tccggcctgc ggcgggagcc gagggcggcc
     1081 gacgacttcg acgacgacga gggcgaggag gaggacgagg cggcggcggc agcggcggcg
     1141 gcagcgatcg gctaccgagg tccatatact tttgttcagc aacatctcat gattggcacc
     1201 gatcctcgaa caattcttaa agatttatta ccagaaacaa ttcctccacc tgagctggat
     1261 gatatgacgc tgtggcagat tgttattaat atcctttcag aaccaccaaa gcggaaaaaa
     1321 agaaaagata tcaatacaat tgaagatgct gtgaagttac tgcaggagtg taaaaagata
     1381 atagttctga ctggagctgg gctcgagcgg ccgcatcgtg actgactgac gatctgcctc
     1441 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     1501 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     1561 ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt gtataattct
     1621 tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     1681 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     1741 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     1801 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     1861 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     1921 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     1981 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     2041 ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc
     2101 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     2161 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     2221 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     2281 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     2341 aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta
     2401 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     2461 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     2521 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     2581 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     2641 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     2701 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     2761 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     2821 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     2881 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     2941 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     3001 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     3061 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     3121 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     3181 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     3241 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     3301 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     3361 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     3421 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     3481 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     3541 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     3601 gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg
     3661 tgcggtattt cacaccgcat aaattccgac accatcgaat ggcgcaaaac ctttcgcggt
     3721 atggcatgat agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg
     3781 ttatacgatg tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac
     3841 caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg
     3901 aattacattc ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt gctgattggc
     3961 gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct
     4021 cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa
     4081 gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg tcagtgggct gatcattaac
     4141 tatccgctgg atgaccagga tgccattgct gtggaagctg cctgcactaa tgttccggcg
     4201 ttatttcttg atgtctctga ccagacaccc atcaacagta ttattttctc ccatgaagac
     4261 ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc accagcaaat cgcgctgtta
     4321 gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc
     4381 actcgcaatc aaattcagcc gatagcggaa cgggaaggcg actggagtgc catgtccggt
     4441 tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat gctggttgcc
     4501 aacgatcaga tggcgctggg cgcaatgcgc gccattaccg agtccgggct gcgcgttggt
     4561 gcggatatct cggtagtggg atacgacgat accgaagaca gctcatgtta tatcccgccg
     4621 ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg
     4681 caactctctc agggccaggc ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa
     4741 agaaaaacca ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     4801 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     4861 taatgtaagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     4921 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
     4981 ttacggattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     5041 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     5101 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc tttgcctggt
     5161 ttccggcacc agaagcggtg ccggaaagct ggctggagtg cgatcttcct gaggccgata
     5221 ctgtcgtcgt cccctcaaac tggcagatgc acggttacga tgcgcccatc tacaccaacg
     5281 tgacctatcc cattacggtc aatccgccgt ttgttcccac ggagaatccg acgggttgtt
     5341 actcgctcac atttaatgtt gatgaaagct ggctacagga aggccagacg cgaattattt
     5401 ttgatggcgt tggaattagc ttatcgactg cacggtgcac caatgcttct ggcgtcaggc
     5461 agccatcgga agctgtggta tggctgtgca ggtcgtaaat cactgcataa ttcgtgtcgc
     5521 tcaaggcgca ctcccgttct ggataatgtt ttttgcgccg acatcataac ggttctggca
     5581 aatattctga aatgagctg
//
