LOCUS       sequence-469338-        5022 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo-YTK; ConLS/R1 cassette; NS3 systems; 
            pTDH3-LexA-NES-NS3-V2-NLS-VP48-2XOaf1-tADH1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5022)
  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 5022)
  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: pWN104
FEATURES             Location/Qualifiers
     source          1..5022
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(103..894,895..963))
                     /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     726..745
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(964..1068)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1171..1759
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1660..1679
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2078..2744
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     2614..2637
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     CDS             join(2751..3002,3003..3356)
                     /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             3372..3398
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             4266..4292
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4299..4319
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      4621..4808
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4676..4700)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
ORIGIN
        1 gcggccgcga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
       61 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
      121 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
      181 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
      241 gcgggaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
      301 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
      361 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
      421 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
      481 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
      541 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
      601 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
      661 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
      721 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
      781 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
      841 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
      901 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
      961 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     1021 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     1081 aaaagtgcca cctgtcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     1141 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     1201 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     1261 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     1321 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     1381 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     1441 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     1501 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     1561 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     1621 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     1681 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     1741 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     1801 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtgc
     1861 ggccgcccct gaattcgcat ctagatggta gagccacaaa cagccggtac aagcaacgat
     1921 ctccaggacc atctgaatca tgcgcggatg acacgaactc acgacggcga tcacagacat
     1981 taacccacag tacagacact gcgacaacgt ggcaattcgt cgcaataccg tctcactgaa
     2041 ctggccgata attgcagacg aacgcagttc gagtttatca ttatcaatac tgccatttca
     2101 aagaatacgt aaataattaa tagtagtgat tttcctaact ttatttagtc aaaaaattag
     2161 ccttttaatt ctgctgtaac ccgtacatgc ccaaaatagg gggcgggtta cacagaatat
     2221 ataacatcgt aggtgtctgg gtgaacagtt tattcctggc atccactaaa tataatggag
     2281 cccgcttttt aagctggcat ccagaaaaaa aaagaatccc agcaccaaaa tattgttttc
     2341 ttcaccaacc atcagttcat aggtccattc tcttagcgca actacagaga acaggggcac
     2401 aaacaggcaa aaaacgggca caacctcaat ggagtgatgc aacctgcctg gagtaaatga
     2461 tgacacaagg caattgaccc acgcatgtat ctatctcatt ttcttacacc ttctattacc
     2521 ttctgctctc tctgatttgg aaaaagctga aaaaaaaggt tgaaaccagt tccctgaaat
     2581 tattccccta cttgactaat aagtatataa agacggtagg tattgattgt aattctgtaa
     2641 atctatttct taaacttctt aaattctact tttatagtta gtcttttttt tagttttaaa
     2701 acaccaagaa cttagtttcg aataaacaca cataaacaaa caaaagatct atgaaggcac
     2761 taacggcgcg tcaacaggaa gtatttgatc taatcagaga tcacatttct cagacgggca
     2821 tgccacctac gagagcggaa attgcgcaga ggctggggtt tcgttcacca aacgcggccg
     2881 aggaacacct gaaggctcta gcaagaaaag gtgtcattga gatagtctcc ggtgcctcca
     2941 ggggaatcag gcttctacaa gaggaggagg aaggactgcc acttgtagga agggtcgctg
     3001 caggggagcc tctattggcg cagcagcata tcgaaggtca ttatcaggtc gatccatctt
     3061 tgttcaagcc caatgccgat ttcttacttc gtgtgagtgg aatgagtatg aaagatatag
     3121 gcattatgga tggggatcta cttgctgttc ataaaacaca agatgtgaga aatggtcagg
     3181 tagttgtagc aaggatcgat gacgaagtca cagttaaaag gctaaagaag caaggcaata
     3241 aagtagagtt attacccgag aactcagaat ttaagcctat tgtggtcgac ctacgtcaac
     3301 agagcttcac catagaaggt ctggccgtcg gtgttatccg taacggtgat tggctggggt
     3361 cagggtcacc tctgcctccg ttagaaagac taacgctggg ttctggggcg agcgcgggca
     3421 tggcgtcaat gactgggggc cagcaaatgg ggagtacgtt gtaccaagag tttgacgaga
     3481 tggaagagtg ttcacaacat ggcaagaaga agggtgacat cgacacgtac agatacatag
     3541 gctctagtgg gacaggctgc gtagtaatcg tgggcagaat tgtcctaagc ggatccggaa
     3601 ccagtgcgcc tatcaccgcc tacgcacaac aaaccagggg actattgggc tgtatcataa
     3661 caagcctaac tggaagagac aaaaatcaag tcgagggcga ggttcagatc gtatctacgg
     3721 caactcagac ctttcttgcc acgtgcataa acggcgtgtg ctgggccgta tatcatgggg
     3781 ctggcacaag aacgatagcg agtcctaagg gtcctgttat tcaaatgtat accaacgtag
     3841 accaggatct agtcggctgg cccgcgcccc aaggctctag gagtcttacc ccttgcacgt
     3901 gcgggtcctc agacttatac ctggtaacga ggcacgcaga tgttatacct gtgcgtagaa
     3961 ggggtgacag tagaggatct cttttatccc cccgtcccat tagctactta aagggtagct
     4021 ctggggggcc gttgttatgt ccggcaggac atgccgtcgg gttattcaga gcggctgttt
     4081 gtacgagggg tgttgcaaaa gcagtagatt ttataccggt tgagaactta gagacaacga
     4141 tgaggtcccc ggtttttaca gacaactcat ctccacctgc tgttacactg acccacccca
     4201 tcaccaaaat tgacagggag gttctctacc aggaatttga cgaaatggag gagtgctcac
     4261 aacactatcc atacgacgta ccagattatg ctggatcccc caagaagaag cgtaaagtgc
     4321 cggcggatgc tctagacgat ttcgatctgg atatgctacc agccgatgca cttgacgatt
     4381 tcgacttaga catgcttccg gccgacgctc tagatgactt cgacttagat atgctcggtt
     4441 ccggcgcgaa caatacacca tttccgggat attttggggg ccttgattta ttcgactacg
     4501 acttcctttt cggcaacgac tttgctggct caggtgcgaa taatactcct tttccaggtt
     4561 attttggagg tttggattta tttgattatg actttttgtt tggcaatgac tttgcttggc
     4621 gcgaatttct tatgatttat gatttttatt attaaataag ttataaaaaa aataagtgta
     4681 tacaaatttt aaagtgactc ttaggtttta aaacgaaaat tcttattctt gagtaactct
     4741 ttcctgtagg tcaggttgct ttctcaggta tagcatgagg tcgctcttat tgaccacacc
     4801 tctaccggca tgccgagcaa atgcctgcaa atcgctcccc atttcgctgc caatgagacg
     4861 acggggtcat cacggctcat catgcgccaa acaaatgtgt gcaatacacg ctcggatgac
     4921 tgcatgatga ccgcactgac tggggacagc agatccacct aagcctgtga gagaagcaga
     4981 cacccgacag atcaaggcag ttaactagtg cactgcagta ca
//
