LOCUS       sequence-137164-        6737 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 6737)
  AUTHORS   Hallows WC, Lee S, Denu JM
  TITLE     Sirtuins deacetylate and activate mammalian acetyl-CoA synthetases.
  JOURNAL   Proc Natl Acad Sci U S A. 2006 Jul 5. 103(27):10230-5.
  PUBMED    16790548
REFERENCE   2  (bases 1 to 6737)
  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: "Proc Natl 
            Acad Sci U S A. 2006 Jul 5. 103(27):10230-5."
COMMENT     Sequence Label: SIRT1.1
FEATURES             Location/Qualifiers
     source          1..6737
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(19..33,34..39,40..56,57..63)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    39..55
                     /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)."
     protein_bind    71..87
                     /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)."
     RBS             106..117
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             136..153
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2203..2297
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             2341..3000
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2407..2426)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      3065..3151
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3244..4326)
                     /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     4288..4307
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4327..4404)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     primer_bind     complement(4488..4510)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4596..4736
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4651..4670
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4751..4768)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4922..5510)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5002..5021)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5681..6472,6473..6541))
                     /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     6304..6323
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6542..6646)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6714..6732
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cgtcttcacc tcgagaaatc ataaaaaatt tatttgcttt gtgagcggat aacaattata
       61 atagattcaa ttgtgagcgg ataacaattt cacacagaat tcattaaaga ggagaaatta
      121 actatgagag gatcgcatca ccatcaccat cacggatcca tggcggacga ggcggcggcg
      181 gcaggcgggg agcaagaggc ccaggcgact gcggcggctg gggaaggaga caatgggccg
      241 ggcctgcagg gcccatctcg ggagccaccg ctggccgaca acttgtacga cgaagacgac
      301 gacgacgagg gcgaggagga ggaagaggcg gcggcggcgg cgattgggta ccgagataac
      361 cttctgttcg gtgatgaaat tatcactaat ggttttcatt cctgtgaaag tgatgaggag
      421 gatagagcct cacatgcaag ctctagtgac tggactccaa ggccacggat aggtccatat
      481 acttttgttc agcaacatct tatgattggc acagatcctc gaacaattct taaagattta
      541 ttgccggaaa caatacctcc acctgagttg gatgatatga cactgtggca gattgttatt
      601 aatatccttt cagaaccacc aaaaaggaaa aaaagaaaag atattaatac aattgaagat
      661 gctgtgaaat tactgcaaga gtgcaaaaaa attatagttc taactggagc tggggtgtct
      721 gtttcatgtg gaatacctga cttcaggtca agggatggta tttatgctcg ccttgctgta
      781 gacttcccag atcttccaga tcctcaagcg atgtttgata ttgaatattt cagaaaagat
      841 ccaagaccat tcttcaagtt tgcaaaggaa atatatcctg gacaattcca gccatctctc
      901 tgtcacaaat tcatagcctt gtcagataag gaaggaaaac tacttcgcaa ctatacccag
      961 aacatagaca cgctggaaca ggttgcggga atccaaagga taattcagtg tcatggttcc
     1021 tttgcaacag catcttgcct gatttgtaaa tacaaagttg actgtgaagc tgtacgagga
     1081 gatattttta atcaggtagt tcctcgatgt cctaggtgcc cagctgatga accgcttgct
     1141 atcatgaaac cagagattgt gttttttggt gaaaatttac cagaacagtt tcatagagcc
     1201 atgaagtatg acaaagatga agttgacctc ctcattgtta ttgggtcttc cctcaaagta
     1261 agaccagtag cactaattcc aagttccata ccccatgaag tgcctcagat attaattaat
     1321 agagaacctt tgcctcatct gcattttgat gtagagcttc ttggagactg tgatgtcata
     1381 attaatgaat tgtgtcatag gttaggtggt gaatatgcca aactttgctg taaccctgta
     1441 aagctttcag aaattactga aaaacctcca cgaacacaaa aagaattggc ttatttgtca
     1501 gagttgccac ccacacctct tcatgtttca gaagactcaa gttcaccaga aagaacttca
     1561 ccaccagatt cttcagtgat tgtcacactt ttagaccaag cagctaagag taatgatgat
     1621 ttagatgtgt ctgaatcaaa aggttgtatg gaagaaaaac cacaggaagt acaaacttct
     1681 aggaatgttg aaagtattgc tgaacagatg gaaaatccgg atttgaagaa tgttggttct
     1741 agtactgggg agaaaaatga aagaacttca gtggctggaa cagtgagaaa atgctggcct
     1801 aatagagtgg caaaggagca gattagtagg cggcttgatg gtaatcagta tctgtttttg
     1861 ccaccaaatc gttacatttt ccatggcgct gaggtatatt cagactctga agatgacgtc
     1921 ttatcctcta gttcttgtgg cagtaacagt gatagtggga catgccagag tccaagttta
     1981 gaagaaccca tggaggatga aagtgaaatt gaagaattct acaatggctt agaagatgag
     2041 cctgatgttc cagagagagc tggaggagct ggatttggga ctgatggaga tgatcaagag
     2101 gcaattaatg aagctatatc tgtgaaacag gaagtaacag acatgaacta tccatcaaac
     2161 aaatcatgtc gacctgcagc caagcttaat tagctgagct tggactcctg ttgatagatc
     2221 cagtaatgac ctcagaactc catctggatt tgttcagaac gctcggttgc cgccgggcgt
     2281 tttttattgg tgagaatcca agctagcttg gcgagatttt caggagctaa ggaagctaaa
     2341 atggagaaaa aaatcactgg atataccacc gttgatatat cccaatggca tcgtaaagaa
     2401 cattttgagg catttcagtc agttgctcaa tgtacctata accagaccgt tcagctggat
     2461 attacggcct ttttaaagac cgtaaagaaa aataagcaca agttttatcc ggcctttatt
     2521 cacattcttg cccgcctgat gaatgctcat ccggaatttc gtatggcaat gaaagacggt
     2581 gagctggtga tatgggatag tgttcaccct tgttacaccg ttttccatga gcaaactgaa
     2641 acgttttcat cgctctggag tgaataccac gacgatttcc ggcagtttct acacatatat
     2701 tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg gtttattgag
     2761 aatatgtttt tcgtctcagc caatccctgg gtgagtttca ccagttttga tttaaacgtg
     2821 gccaatatgg acaacttctt cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc
     2881 gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga tggcttccat
     2941 gtcggcagaa tgcttaatga attacaacag tactgcgatg agtggcaggg cggggcgtaa
     3001 tttttttaag gcagttattg gtgcccttaa acgcctgggg taatgactct ctagcttgag
     3061 gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt
     3121 gtcggtgaac gctctcctga gtaggacaaa tccgccctct agattacgtg cagtcgatga
     3181 taagctgtca aacatgagaa ttgtgcctaa tgagtgagct aacttacatt aattgcgttg
     3241 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     3301 caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag
     3361 tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg
     3421 gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat
     3481 ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatatccg caccaacgcg
     3541 cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag
     3601 catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat
     3661 ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt
     3721 atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc
     3781 gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg
     3841 ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac
     3901 attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat
     3961 cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc
     4021 gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat
     4081 cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag
     4141 caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc
     4201 catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac
     4261 gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg
     4321 tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa
     4381 ggttttgcac cattcgatgg tgtcggaatt tcgggcagcg ttgggtcctg gccacgggtg
     4441 cgcatgatct agagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     4501 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     4561 cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc
     4621 gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     4681 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct
     4741 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     4801 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     4861 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     4921 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     4981 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     5041 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     5101 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     5161 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     5221 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     5281 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     5341 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     5401 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     5461 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     5521 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     5581 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     5641 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     5701 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     5761 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     5821 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     5881 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     5941 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     6001 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     6061 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     6121 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     6181 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     6241 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     6301 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     6361 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     6421 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     6481 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     6541 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     6601 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     6661 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     6721 gtatcacgag gcccttt
//
