LOCUS       sequence-313742-        5830 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 5830)
  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 5830)
  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: ACS1
FEATURES             Location/Qualifiers
     source          1..5830
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..15,16..21,22..38,39..45)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    21..37
                     /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    53..69
                     /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             88..99
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             118..135
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(2319..3401)
                     /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     3363..3382
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3402..3479)
                     /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(3563..3585)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3671..3811
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3726..3745
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3826..3843)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3997..4585)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4077..4096)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4756..5547,5548..5616))
                     /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     5379..5398
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5617..5721)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5789..5807
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcataaaaaa tttatttgct ttgtgagcgg ataacaatta taatagattc aattgtgagc
       61 ggataacaat ttcacacaga attcattaaa gaggagaaat taactatgag aggatcgcat
      121 caccatcacc atcacggatc cgcatgcgag ctcatggggc ttcccgagga gcggcgcaag
      181 agcggcagtg gaagccgggc tcgtgaggag accggcgccg agggccgggt gcggggttgg
      241 tccccgccgc cggaggtcag acgctcggcg cacgtcccct ctctgcagcg ctaccgcgag
      301 ctgcaccggc gttctgtgga ggagccacgg gagttttggg gaaacattgc caaggaattt
      361 tactggaaaa ccgcatgccc tggcccattc ctccagtaca actttgatgt gactaaaggg
      421 aaaatattca ctgagtggat gaaaggagca actacaaaca tctgctacaa cgtgctggat
      481 cgaaatgtcc atgagaaaaa acttggcgac aaagttgctt tttactggga gggcaatgag
      541 ccaggggaga ccaccaagat cacataccgt gaactcctgg tccaggtgtg tcagttcagc
      601 aatgttctcc gtaaacaggg cattcagaag ggtgaccgag tggccatcta catgcctatg
      661 atcttggaac ttgtggtggc tatgctggca tgcgctcgcc ttggagcttt gcactccatt
      721 gtgtttgcag gcttctctgc agagtctctc tgtgaaagga tcttggattc cagttgctgc
      781 ctgctcatca ctacagatgc cttctacagg ggggaaaaac ttgtgaacct gaaggagctg
      841 gctgatgagt ccttggagaa gtgccgagag aagggcttcc cagtgagatg ctgcattgtg
      901 gtcaaacatc tggggcgggc agagctgggc atgaatgact cccccagcca gtccccacca
      961 gttaagaggc catgtccaga tgtccagatc tgctggaacg aaggggttga cttatggtgg
     1021 catgaactca tgcagcaggc aggagacgag tgtgagcctg agtggtgtga tgctgaggac
     1081 ccactcttca tcttgtacac cagtggctcc acaggcaaac ctaagggtgt ggtgcacaca
     1141 attggaggct atatgctcta tgtggctaca actttcaagt atgtgtttga tttccacccg
     1201 gaagatgtgt tctggtgcac agcagacatc ggctggatca ctggtcattc ctatgtcacc
     1261 tatgggccac tggctaatgg tgccactagt gttttgtttg aggggatccc cacataccca
     1321 gatgaagggc gcttgtggag cattgtggac aaatacaagg tgaccaagtt ctacacggca
     1381 ccaacagcca tccggatgct catgaagttt ggagatgatc ctgtcaccaa gcatagccgg
     1441 gcatccctgc aggtgctggg cacagtaggt gaacccatca accctgaagc ctggctatgg
     1501 taccaccggg tagtaggttc ccagcgctgc cccattgtag acaccttctg gcaaacagaa
     1561 acaggtggcc atatgctgac ccctctccct ggcgccacac ccatgaaacc tggttctgct
     1621 tctttcccat tcttcggtgt agcgcctgca atcctgaatg agtccgggga ggagctggaa
     1681 ggggaagctg aaggttatct ggtgttcaag cagccctggc cagggatcat gcgcacagtc
     1741 tatgggaacc acacacggtt tgagaccacc tactttaaga agttccctgg ctactatgtg
     1801 accggagatg gctgccgacg ggatcaggat ggctattact ggatcacggg caggattgat
     1861 gacatgctca atgtgtctgg acatctcctg agtacagcag aggtggaatc ggcacttgtg
     1921 gaacacgagg ctgtcgcaga ggcagctgtg gtgggccacc ctcatcctgt gaagggcgaa
     1981 tgcctctact gctttgttac cttgtgtgat ggccatacct tcagccccac actcacagag
     2041 gaactcaaga agcagattag agaaaagatt ggccccattg ccacaccaga ctacatccag
     2101 aatgcacctg gcttgcctaa aacacgctca gggaaaatca tgaggcgagt tctccggaag
     2161 attgctcaga atgaccatga cctgggggat acatctacgg tggctgaccc atctgtcatc
     2221 aaccatctct tcagtcaccg ctgcctgacc acccagaagc tgtcaaacat gagaattgtg
     2281 cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg
     2341 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     2401 gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc
     2461 ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg
     2521 cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg
     2581 tcgtatccca ctaccgagat atccgcacca acgcgcagcc cggactcggt aatggcgcgc
     2641 attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca
     2701 ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc
     2761 gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc
     2821 gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc
     2881 agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt
     2941 gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca
     3001 atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga
     3061 agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc
     3121 acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg
     3181 tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt
     3241 tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc
     3301 gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca
     3361 ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga
     3421 ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcaccattc gatggtgtcg
     3481 gaatttcggg cagcgttggg tcctggccac gggtgcgcat gatctagagc tgcctcgcgc
     3541 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     3601 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     3661 ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat actggcttaa
     3721 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     3781 cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc tcactgactc
     3841 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3901 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3961 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4021 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4081 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4141 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4201 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4261 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4321 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4381 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     4441 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4501 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4561 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4621 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4681 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4741 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4801 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4861 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     4921 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     4981 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5041 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5101 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5161 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5221 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5281 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5341 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5401 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     5461 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     5521 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     5581 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     5641 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     5701 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     5761 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca
     5821 cctcgagaaa
//
