LOCUS       sequence-469336-        4992 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo-YTK; ConLS/R1 cassette; NS3 systems; 
            pTDH3-LexA-NLS-NS3-V1-VP48-2XOaf1-tADH1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4992)
  AUTHORS   O'Laughlin R, Tran Q, Lezia A, Ngamkanjanarat W, Emmanuele P, Hao N,
            Hasty J
  TITLE     A Standardized Set of MoClo-Compatible Inducible Promoter Systems 
            for Tunable Gene Expression in Yeast.
  JOURNAL   ACS Synth Biol. 2024 Jan 19;13(1):85-102. doi: 
            10.1021/acssynbio.3c00184. Epub 2023 Dec 11.
  PUBMED    38079574
REFERENCE   2  (bases 1 to 4992)
  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: 
            "10.1021/acssynbio.3c00184"; journalName: "ACS Synth Biol"; date: 
            "2024-01-19- 19"; volume: "13"; issue: "1"; pages: "85-102"
COMMENT     Sequence Label: pAL008
FEATURES             Location/Qualifiers
     source          1..4992
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        908..1574
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     1444..1467
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     CDS             join(1581..1832,1833..2186)
                     /codon_start=1
                     /gene="lexA"
                     /product="E. coli DNA-binding protein that represses genes 
                     involved in the SOS response to DNA damage"
                     /label=LexA
                     /note="The N-terminal DNA-binding domain recognizes the 
                     16-bp palindromic sequence 5'-CTGTATATATATACAG-3'."
                     /translation="MKALTARQQEVFDLIRDHISQTGMPPTRAEIAQRLGFRSPNAAEE
                     HLKALARKGVIEIVSGASRGIRLLQEEEEGLPLVGRVAAGEPLLAQQHIEGHYQVDPSL
                     FKPNADFLLRVSGMSMKDIGIMDGDLLAVHKTQDVRNGQVVVARIDDEVTVKRLKKQGN
                     KVELLPENSEFKPIVVDLRQQSFTIEGLAVGVIRNGDWL"
     CDS             2202..2222
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3084..3110
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     terminator      3421..3608
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3476..3500)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(join(3925..4716,4717..4785))
                     /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     4548..4567
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4786..4890)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtgc ggccgcccct gaattcgcat ctagatggta
      721 gagccacaaa cagccggtac aagcaacgat ctccaggacc atctgaatca tgcgcggatg
      781 acacgaactc acgacggcga tcacagacat taacccacag tacagacact gcgacaacgt
      841 ggcaattcgt cgcaataccg tctcactgaa ctggccgata attgcagacg aacgcagttc
      901 gagtttatca ttatcaatac tgccatttca aagaatacgt aaataattaa tagtagtgat
      961 tttcctaact ttatttagtc aaaaaattag ccttttaatt ctgctgtaac ccgtacatgc
     1021 ccaaaatagg gggcgggtta cacagaatat ataacatcgt aggtgtctgg gtgaacagtt
     1081 tattcctggc atccactaaa tataatggag cccgcttttt aagctggcat ccagaaaaaa
     1141 aaagaatccc agcaccaaaa tattgttttc ttcaccaacc atcagttcat aggtccattc
     1201 tcttagcgca actacagaga acaggggcac aaacaggcaa aaaacgggca caacctcaat
     1261 ggagtgatgc aacctgcctg gagtaaatga tgacacaagg caattgaccc acgcatgtat
     1321 ctatctcatt ttcttacacc ttctattacc ttctgctctc tctgatttgg aaaaagctga
     1381 aaaaaaaggt tgaaaccagt tccctgaaat tattccccta cttgactaat aagtatataa
     1441 agacggtagg tattgattgt aattctgtaa atctatttct taaacttctt aaattctact
     1501 tttatagtta gtcttttttt tagttttaaa acaccaagaa cttagtttcg aataaacaca
     1561 cataaacaaa caaaagatct atgaaggctt taactgcaag acaacaggaa gtttttgatt
     1621 tgataagaga tcatatatct caaactggaa tgccaccaac aagagctgaa atagctcaaa
     1681 gattaggatt tagatctcca aacgctgcag aagaacattt gaaagctttg gcaaggaaag
     1741 gtgtcatcga aattgtctct ggtgcttcaa gaggtattag actattacaa gaagaggaag
     1801 aaggattgcc attggttggt agagttgctg ctggtgaacc attattagca cagcaacata
     1861 ttgagggtca ttatcaagta gatccatctt tattcaaacc aaatgctgat tttttgttga
     1921 gagtaagtgg tatgtctatg aaagatattg gtatcatgga tggtgactta ttggctgttc
     1981 ataaaactca agatgttaga aacggtcaag ttgtcgttgc cagaattgat gatgaagtca
     2041 ctgttaaaag attgaagaaa caaggtaata aagtggaatt gttaccagaa aattctgaat
     2101 ttaagccaat tgttgttgat ttaaggcaac aatcttttac tattgaaggt ttagctgtcg
     2161 gtgttatcag aaatggtgat tggttaggtt caggttctcc tccaaaaaaa aagagaaaag
     2221 ttggatctgg ttctggggcg agcgcgggca tggcgtcaat gactgggggc cagcaaatgg
     2281 ggagtacgga ggatgtagtt tgttgtcatt caatatacgg caagaagaag ggtgacatcg
     2341 acacgtacag atacataggc tctagtggga caggctgcgt agtaatcgtg ggcagaattg
     2401 tcctaagcgg atccggaacc agtgcgccta tcaccgccta cgcacaacaa accaggggac
     2461 tattgggctg tatcataaca agcctaactg gaagagacaa aaatcaagtc gagggcgagg
     2521 ttcagatcgt atctacggca actcagacct ttcttgccac gtgcataaac ggcgtgtgct
     2581 gggccgtata tcatggggct ggcacaagaa cgatagcgag tcctaagggt cctgttattc
     2641 aaatgtatac caacgtagac caggatctag tcggctggcc cgcgccccaa ggctctagga
     2701 gtcttacccc ttgcacgtgc gggtcctcag acttatacct ggtaacgagg cacgcagatg
     2761 ttatacctgt gcgtagaagg ggtgacagta gaggatctct tttatccccc cgtcccatta
     2821 gctacttaaa gggtagctct ggggggccgt tgttatgtcc ggcaggacat gccgtcgggt
     2881 tattcagagc ggctgtttgt acgaggggtg ttgcaaaagc agtagatttt ataccggttg
     2941 agaacttaga gacaacgatg aggtccccgg tttttacaga caactcatct ccacctgctg
     3001 ttacactgac ccaccccatc accaaaattg acagggaggt tctctaccag gaatttgacg
     3061 aaatggagga gtgctcacaa cactatccat acgacgtacc agattatgct ggatccccgg
     3121 cggatgctct agacgatttc gatctggata tgctaccagc cgatgcactt gacgatttcg
     3181 acttagacat gcttccggcc gacgctctag atgacttcga cttagatatg ctcggttccg
     3241 gcgcgaacaa tacaccattt ccgggatatt ttgggggcct tgatttattc gactacgact
     3301 tccttttcgg caacgacttt gctggctcag gtgcgaataa tactcctttt ccaggttatt
     3361 ttggaggttt ggatttattt gattatgact ttttgtttgg caatgacttt gcttaatggc
     3421 gcgaatttct tatgatttat gatttttatt attaaataag ttataaaaaa aataagtgta
     3481 tacaaatttt aaagtgactc ttaggtttta aaacgaaaat tcttattctt gagtaactct
     3541 ttcctgtagg tcaggttgct ttctcaggta tagcatgagg tcgctcttat tgaccacacc
     3601 tctaccggca tgccgagcaa atgcctgcaa atcgctcccc atttcgctgc caatgagacg
     3661 acggggtcat cacggctcat catgcgccaa acaaatgtgt gcaatacacg ctcggatgac
     3721 tgcatgatga ccgcactgac tggggacagc agatccacct aagcctgtga gagaagcaga
     3781 cacccgacag atcaaggcag ttaactagtg cactgcagta cagcggccgc gattatcaaa
     3841 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     3901 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     3961 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     4021 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgggacc cacgctcacc
     4081 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     4141 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     4201 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     4261 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     4321 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     4381 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     4441 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     4501 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     4561 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     4621 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     4681 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     4741 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     4801 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     4861 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgtcat
     4921 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     4981 caaaggatct tc
//
