LOCUS       sequence-468711-        3911 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo-YTK; ConLS/R1 cassette; rtTA systems; pTDH3-rtTA-v10-tADH1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3911)
  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 3911)
  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: pRO253
FEATURES             Location/Qualifiers
     source          1..3911
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        212..878
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     748..771
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     CDS             885..1628
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     terminator      1644..1831
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1699..1723)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(join(2148..2939,2940..3008))
                     /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     2771..2790
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3009..3113)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3216..3804
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3705..3724
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccctgaattc gcatctagat ggtagagcca caaacagccg gtacaagcaa cgatctccag
       61 gaccatctga atcatgcgcg gatgacacga actcacgacg gcgatcacag acattaaccc
      121 acagtacaga cactgcgaca acgtggcaat tcgtcgcaat accgtctcac tgaactggcc
      181 gataattgca gacgaacgca gttcgagttt atcattatca atactgccat ttcaaagaat
      241 acgtaaataa ttaatagtag tgattttcct aactttattt agtcaaaaaa ttagcctttt
      301 aattctgctg taacccgtac atgcccaaaa tagggggcgg gttacacaga atatataaca
      361 tcgtaggtgt ctgggtgaac agtttattcc tggcatccac taaatataat ggagcccgct
      421 ttttaagctg gcatccagaa aaaaaaagaa tcccagcacc aaaatattgt tttcttcacc
      481 aaccatcagt tcataggtcc attctcttag cgcaactaca gagaacaggg gcacaaacag
      541 gcaaaaaacg ggcacaacct caatggagtg atgcaacctg cctggagtaa atgatgacac
      601 aaggcaattg acccacgcat gtatctatct cattttctta caccttctat taccttctgc
      661 tctctctgat ttggaaaaag ctgaaaaaaa aggttgaaac cagttccctg aaattattcc
      721 cctacttgac taataagtat ataaagacgg taggtattga ttgtaattct gtaaatctat
      781 ttcttaaact tcttaaattc tacttttata gttagtcttt tttttagttt taaaacacca
      841 agaacttagt ttcgaataaa cacacataaa caaacaaaag atctatgtcc cgtcttgata
      901 agagtaaggt tatcaactca gccctagagc tgctgaacgg cgtcggaatc gaggggctta
      961 ctaccagaaa actagcacag aaactgggtg tagagcagcc cacgctttac tggcatgtta
     1021 aaaataagag agcactatta gacgcgttgc cgatcgaaat gttggatagg catcacacac
     1081 actcatgccc tttagaagga gaaagttggc aggattttct gagaaataat gcgaagtctt
     1141 atcgttgtgc gttactttcc cacagggacg gtgctaaagt acatttggga actagaccaa
     1201 ctgaaaaaca atatgagaca ttggagaacc aactggcctt cttatgccaa cagggtttca
     1261 gtcttgagaa cgccctttac gccctttcag cggttgggca cttcactctt ggttgcgtct
     1321 tggaagagca ggagcatcaa gtagcaaaag aggagcgtga aacccccaca acagactcca
     1381 tgccccctct attgaagcaa gctatcgaat tattcgaccg tcaaggagcg gaaccagcat
     1441 ttttatttgg cttagaattg attatttgtg ggttagaaaa acaactaaag tgtgagagcg
     1501 gtggaccgac agacgctctt gatgattttg acttggatat gttacccgcc gacgctttgg
     1561 acgacttcga tttggatatg ctgcctgcgg atgcactaga tgatttcgat ctggacatgt
     1621 taccgggtgg atcctaactc gaggcgaatt tcttatgatt tatgattttt attattaaat
     1681 aagttataaa aaaaataagt gtatacaaat tttaaagtga ctcttaggtt ttaaaacgaa
     1741 aattcttatt cttgagtaac tctttcctgt aggtcaggtt gctttctcag gtatagcatg
     1801 aggtcgctct tattgaccac acctctaccg gcatgccgag caaatgcctg caaatcgctc
     1861 cccatttcgc tgccaatgag acgacggggt catcacggct catcatgcgc caaacaaatg
     1921 tgtgcaatac acgctcggat gactgcatga tgaccgcact gactggggac agcagatcca
     1981 cctaagcctg tgagagaagc agacacccga cagatcaagg cagttaacta gtgcactgca
     2041 gtacagcggc cgcgattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     2101 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     2161 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     2221 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     2281 ataccgcggg acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     2341 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     2401 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     2461 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     2521 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     2581 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     2641 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     2701 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     2761 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     2821 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     2881 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     2941 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     3001 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     3061 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     3121 ccccgaaaag tgccacctgt catgaccaaa atcccttaac gtgagttttc gttccactga
     3181 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     3241 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     3301 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     3361 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     3421 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     3481 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     3541 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     3601 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     3661 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     3721 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     3781 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     3841 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     3901 cgtgcggccg c
//
