LOCUS       sequence-468709-        3931 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo-YTK; ConLS/R1 cassette; rtTA systems; pRPL18B-rtTA-v10-tADH1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3931)
  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 3931)
  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: pRO229
FEATURES             Location/Qualifiers
     source          1..3931
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             906..1649
                     /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      1665..1852
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1720..1744)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(join(2169..2960,2961..3029))
                     /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     2792..2811
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3030..3134)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3237..3825
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3726..3745
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cccctgaatt cgcatctaga tggtagagcc acaaacagcc ggtacaagca acgatctcca
       61 ggaccatctg aatcatgcgc ggatgacacg aactcacgac ggcgatcaca gacattaacc
      121 cacagtacag acactgcgac aacgtggcaa ttcgtcgcaa taccgtctca ctgaactggc
      181 cgataattgc agacgaacga agaggatgtc caatattttt tttaaggaat aaggatactt
      241 caagactaga ttcccccctg cattcccatc agaaccgtaa accttggcgc tttccttggg
      301 aagtattcaa gaagtgcctt gtccggtttc tgtggctcac aaaccagcgc gcccgatatg
      361 gctttctttt cacttatgaa tgtaccagta cgggacaatt agaacgctcc tgtaacaatc
      421 tctttgcaaa tgtggggtta cattctaacc atgtcacact gctgacgaaa ttcaaagtaa
      481 aaaaaaatgg gaccacgtct tgagaacgat agattttctt tattttacat tgaacagtcg
      541 ttgtctcagc gcgctttatg ttttcattca tacttcatat tataaaataa caaaagaaga
      601 atttcatatt cacgcccaag aaatcaggct gctttccaaa tgcaattgac acttcattag
      661 ccatcacaca aaactctttc ttgctggagc ttcttttaaa aaagacctca gtacaccaaa
      721 cacgttaccc gacctcgtta ttttacgaca actatgataa aattctgaag aaaaaataaa
      781 aaaattttca tacttcttgc ttttatttaa accattgaat gatttctttt gaacaaaact
      841 acctgtttca ccaaaggaaa tagaaagaaa aaatcaatta gaagaaaaca aaaaacaaaa
      901 gatctatgtc ccgtcttgat aagagtaagg ttatcaactc agccctagag ctgctgaacg
      961 gcgtcggaat cgaggggctt actaccagaa aactagcaca gaaactgggt gtagagcagc
     1021 ccacgcttta ctggcatgtt aaaaataaga gagcactatt agacgcgttg ccgatcgaaa
     1081 tgttggatag gcatcacaca cactcatgcc ctttagaagg agaaagttgg caggattttc
     1141 tgagaaataa tgcgaagtct tatcgttgtg cgttactttc ccacagggac ggtgctaaag
     1201 tacatttggg aactagacca actgaaaaac aatatgagac attggagaac caactggcct
     1261 tcttatgcca acagggtttc agtcttgaga acgcccttta cgccctttca gcggttgggc
     1321 acttcactct tggttgcgtc ttggaagagc aggagcatca agtagcaaaa gaggagcgtg
     1381 aaacccccac aacagactcc atgccccctc tattgaagca agctatcgaa ttattcgacc
     1441 gtcaaggagc ggaaccagca tttttatttg gcttagaatt gattatttgt gggttagaaa
     1501 aacaactaaa gtgtgagagc ggtggaccga cagacgctct tgatgatttt gacttggata
     1561 tgttacccgc cgacgctttg gacgacttcg atttggatat gctgcctgcg gatgcactag
     1621 atgatttcga tctggacatg ttaccgggtg gatcctaact cgaggcgaat ttcttatgat
     1681 ttatgatttt tattattaaa taagttataa aaaaaataag tgtatacaaa ttttaaagtg
     1741 actcttaggt tttaaaacga aaattcttat tcttgagtaa ctctttcctg taggtcaggt
     1801 tgctttctca ggtatagcat gaggtcgctc ttattgacca cacctctacc ggcatgccga
     1861 gcaaatgcct gcaaatcgct ccccatttcg ctgccaatga gacgacgggg tcatcacggc
     1921 tcatcatgcg ccaaacaaat gtgtgcaata cacgctcgga tgactgcatg atgaccgcac
     1981 tgactgggga cagcagatcc acctaagcct gtgagagaag cagacacccg acagatcaag
     2041 gcagttaact agtgcactgc agtacagcgg ccgcgattat caaaaaggat cttcacctag
     2101 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     2161 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     2221 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     2281 tctggcccca gtgctgcaat gataccgcgg gacccacgct caccggctcc agatttatca
     2341 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     2401 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     2461 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     2521 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     2581 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     2641 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     2701 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     2761 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     2821 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     2881 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     2941 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     3001 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     3061 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     3121 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg tcatgaccaa aatcccttaa
     3181 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     3241 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     3301 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     3361 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     3421 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     3481 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     3541 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     3601 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     3661 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     3721 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     3781 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     3841 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     3901 tcccctgatt ctgtggataa ccgtgcggcc g
//
