LOCUS       sequence-469368-        6372 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo-YTK URA multigene plasmid with MCS; Tet systems; 
            pRNR2-rtTA-v16-tADH1; tetO7-pLEU2m-MCS-tENO2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6372)
  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 6372)
  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: pAL004
FEATURES             Location/Qualifiers
     source          1..6372
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      2046..2233
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2101..2125)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    2327..2345
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2359..2377
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2390..2408
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2422..2440
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2453..2471
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2485..2503
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2516..2534
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             3200..4003
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     CDS             complement(4645..5460)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(4645..5460)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     5368..5387
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5452..5471
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5685..6273
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6174..6193
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcggccgcca atgctaaatt cgagtgaaac acaggaagat cagaaaatcc tcatttcatc
       61 catattaaca ataatttcaa atgtttattt gcattatttg aaactaggca agacaagcaa
      121 cgaaacgttt ttgaaaattt tgagtatttt caataaattt gtagaggact cagatattga
      181 aaaaaagcta cagcaattaa tacttgataa gaagagtatt gagaagggca acggttcatc
      241 atctcatgga tctgcacatg aacaaacacc agagtcaaac gacgttgaaa ttgaggctac
      301 tgcgccaatt gatgacaata cagacgatga taacaaaccg aagttatctg atgtagaaaa
      361 ggattaaaga tgctaagaga tagtgatgat atttcataaa taatgtaatt ctatatatgt
      421 taattacctt ttttgcgagg catatttatg gtgaaggata agttttgacc atcaaagaag
      481 gttaatgtgg ctgtggtttc agggtccata aagcccacat ggataacatt acccctgaat
      541 tcgcatctag actgaactgg ccgataattg cagacgaacg agtcgaacaa gaagcaggca
      601 aagtttagag cactgcccct ccgcactcaa aaaagaaaaa actaggagga aaataaaatt
      661 ctcaaccaca caaacacata aacacataca aatacaaata caagcttatt tacttgacat
      721 cgcgcgatct tccactattc agcgccgtcc gccctctctc gtgttttttg tttacgcgac
      781 aactatgcga aatccggagc aacgggcaac cgtttgggga aagaccacac ccacgcgcga
      841 tcgccatggc aacgaggtcg cacacgcccc acacccagac ctccctgcga gcgggcatgg
      901 gtacaatgtc cccgttgcca cagacaccac ttcgtagcac agcgcagagc gtagcgtgtt
      961 gttgctgctg acaaaagaaa atttttctta gcaaagcaaa ggaggggaag cacgggcaga
     1021 tagcaccgta ccataccctt ggaaactcga aatgaacgaa gcaggaaatg agagaatgag
     1081 agttttgtag gtatatatag cggtagtgtt tgcgcgttac catcatcttc tggatctatc
     1141 tattgttctt ttcctcatca ctttcccctt tttcgctctt cttcttgtct tttatttctt
     1201 tctttttttt aattgttccc tcgattggct atctaccaaa gaatccaaac ttaatacacg
     1261 tatttatttg tccaattacc agatctatga gtcgtcttga taagtccaag atcataaata
     1321 gcgcactgga attgctgaac ggcgttggta ttgagggtct taccacgaga aagctggcac
     1381 agaagttggg ggtcgaacaa ccgaccttgt actggcatgt caagaataag cgtgcgctat
     1441 tagatgcctt acctatcgag atgctagaca gacatcatac tcactcatgt ccattggagg
     1501 gggagtcttg gcaagacttt cttagaaaca atgcaaagtc atataggtgc gcgctactat
     1561 cacatagaga tggggccaaa gtccatttgg gcaccaggcc gaccgaaaag cagtacgaga
     1621 ctttagagaa ccagctggca ttcctgtgtc aacaaggctt cagtctagag aatgctctgt
     1681 atgctttaag cgcggtcggc cacttcacgc taggttgtgt attagaggaa caagaacacc
     1741 aagtggcaaa agaggaaaga gaaactccaa cgaccgacag catgccacca ctgttgaaac
     1801 aagcaatcga actgtttgat cgtcaagggg ctgaaccagc attcctattc ggtttggaat
     1861 taataatctg cggtctggag aaacaattga agtgcgaaag cggcgggccg acagatgcct
     1921 tagatgactt tgacctagat atgttaccag ccgacgccct tgacgacttt gatctggaca
     1981 tgctgccggc tgacgccttg gatgacttcg acttggatat gttgccaggg ggatcctaac
     2041 tcgaggcgaa tttcttatga tttatgattt ttattattaa ataagttata aaaaaaataa
     2101 gtgtatacaa attttaaagt gactcttagg ttttaaaacg aaaattctta ttcttgagta
     2161 actctttcct gtaggtcagg ttgctttctc aggtatagca tgaggtcgct cttattgacc
     2221 acacctctac cggcatgccg agcaaatgcc tgcaaatcgc tccccatttc gctgccaaac
     2281 cagatgtcaa cacagctaca acgggtagtc catcgttgta ggatactccc tatcagtgat
     2341 agagaaatct atgcggcatc cctatcagtg atagagatcg gtaacggagt ccctatcagt
     2401 gatagagaag tggtgttcag ttccctatca gtgatagaga agtttgacac tatccctatc
     2461 agtgatagag agaacagcaa tgactcccta tcagtgatag agatcaaatg gcgtatccct
     2521 atcagtgata gagataatta acttgtaata ttctaatcaa ttgacaatat tatttaagga
     2581 cctattgttt tttccaatag gtggttagca atcgtcttac tttctaactt ttcttacctt
     2641 ttacatttca gcaatatata tatatatatt tcaaggatat accattctaa gatctgttta
     2701 aacgagctcg ctagcctcga gtctagagtc gacctgcagg taactcgaga gtgcttttaa
     2761 ctaagaatta ttagtctttt ctgcttattt tttcatcata gtttagaaca ctttatatta
     2821 acgaatagtt tatgaatcta tttaggttta aaaattgata cagttttata agttactttt
     2881 tcaaagactc gtgctgtcta ttgcataatg cactggaagg ggaaaaaaaa ggtgcacacg
     2941 cgtggctttt tcttgaattt gcagtttgaa aaatgctgag cagttacaga gatgttacga
     3001 accactagtg cactgcagta cacggtttcc ttgaaatttt tttgattcgg taatctccga
     3061 acagaaggaa gaacgaagga aggagcacag acttagattg gtatatatac gcatatgtag
     3121 tgttgaagaa acatgaaatt gcccagtatt cttaacccaa ctgcacagaa caaaaacgtg
     3181 caggaaacga agataaatca tgtcgaaagc tacatataag gaacgtgctg ctactcatcc
     3241 tagtcctgtt gctgccaagc tatttaatat catgcacgaa aagcaaacaa acttgtgtgc
     3301 ttcattggat gttcgtacca ccaaggaatt actggagtta gttgaagcat taggtcccaa
     3361 aatttgttta ctaaaaacac atgtggatat cttgactgat ttttccatgg agggcacagt
     3421 taagccgcta aaggcattat ccgccaagta caatttttta ctcttcgagg acagaaaatt
     3481 tgctgacatt ggtaatacag tcaaattgca gtactctgcg ggtgtataca gaatagcaga
     3541 atgggcagac attacgaatg cacacggtgt ggtgggccca ggtattgtta gcggtttgaa
     3601 gcaggcggca gaagaagtaa caaaggaacc tagaggcctt ttgatgttag cagaattgtc
     3661 atgcaagggc tccctatcta ctggagaata tactaagggt actgttgaca ttgcgaagag
     3721 cgacaaagat tttgttatcg gctttattgc tcaaagagac atgggtggaa gagatgaagg
     3781 ttacgattgg ttgattatga cacccggtgt gggtttagat gacaagggag atgcattggg
     3841 tcaacagtat agaaccgtgg atgatgtggt ttctacagga tctgacatta ttattgttgg
     3901 aagaggacta tttgcaaagg gaagggatgc taaggtagag ggtgaacgtt acagaaaagc
     3961 aggctgggaa gcatatttga gaagatgcgg ccagcaaaac taaaaaactg tattataagt
     4021 aaatgcatgt atactaaact cacaaattag agcttcaatt taattatatc agttattacc
     4081 cgagtagagc acttgaatcc actgccccgg gaatctcggt cgtaatgatt tctataatga
     4141 cgaaaaaaaa aaaattggaa agaaaaagct tcatggcctt tataaaaagg aactatccaa
     4201 tacctcgcca gaaccaagta acagtatttt acggggcaca aatcaagaac aataagacag
     4261 gactgtaaag atggacgcat tgaactccaa agaacaacaa gagttccaaa aagtagtgga
     4321 acaaaagcaa atgaaggatt tcatgcgttt gtactctaat ctggtagaaa gatgtttcac
     4381 agactgtgtc aatgacttca caacatcaaa gctaaccaat aaggaacaaa catgcatcat
     4441 gaagtgctca gaaaagttct tgaagcatag cgaacgtgta gggcagcgtt tccaagaaca
     4501 aaacgctgcc ttgggacaag gcttgggccg ataaggtgta ctggcgtata tatatctaat
     4561 tatgtatctc tggtgtagcc catttttagc atgtaaatat aaagaccgag cggccgcgtg
     4621 ttacaaccaa ttaaccaatt ctgattagaa aaactcatcg agcatcaaat gaaactgcaa
     4681 tttattcata tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg
     4741 agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc
     4801 gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag
     4861 tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagct tatgcatttc
     4921 tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac
     4981 caaaccgtta ttcattcgtg attgcgcctg agcgaggcga aatacgcgat cgctgttaaa
     5041 aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac
     5101 aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt tcccggggat
     5161 cgcagtggtg agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag
     5221 aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac
     5281 gctacctttg ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcgata
     5341 gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc
     5401 atccatgttg gaatttaatc gcggcctgga gcaagacgtt tcccgttgaa tatggctcat
     5461 aacacccctt gtattactgt ttatgtaagc agacagtttt attgttcatg atgatatatt
     5521 tttatcttgt gcaatgtaac atcagagatt ttgagacaca acgtggcttt gttgaataaa
     5581 tcgaactttt gctgagttga aggatcagtc atgaccaaaa tcccttaacg tgagttttcg
     5641 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     5701 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     5761 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     5821 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     5881 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     5941 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     6001 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6061 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6121 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6181 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6241 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6301 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     6361 gtggataacc gt
//
