LOCUS       sequence-469367-        6333 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo-YTK URA multigene plasmid with MCS; Tet systems; 
            pRPL18B-rtTA3-tADH1; tetO7-pLEU2m-MCS-tENO2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6333)
  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 6333)
  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: pAL003
FEATURES             Location/Qualifiers
     source          1..6333
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     252..271
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      2457..2644
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2512..2536)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    2738..2756
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2770..2788
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2801..2819
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2833..2851
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2864..2882
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2896..2914
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2927..2945
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             3611..4414
                     /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(5056..5871)
                     /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(5056..5871)
                     /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     5779..5798
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5863..5882
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      join(6096..6333,1..351)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
       61 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
      121 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
      181 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
      241 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
      301 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
      361 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
      421 gcgttatccc ctgattctgt ggataaccgt gcggccgcca atgctaaatt cgagtgaaac
      481 acaggaagat cagaaaatcc tcatttcatc catattaaca ataatttcaa atgtttattt
      541 gcattatttg aaactaggca agacaagcaa cgaaacgttt ttgaaaattt tgagtatttt
      601 caataaattt gtagaggact cagatattga aaaaaagcta cagcaattaa tacttgataa
      661 gaagagtatt gagaagggca acggttcatc atctcatgga tctgcacatg aacaaacacc
      721 agagtcaaac gacgttgaaa ttgaggctac tgcgccaatt gatgacaata cagacgatga
      781 taacaaaccg aagttatctg atgtagaaaa ggattaaaga tgctaagaga tagtgatgat
      841 atttcataaa taatgtaatt ctatatatgt taattacctt ttttgcgagg catatttatg
      901 gtgaaggata agttttgacc atcaaagaag gttaatgtgg ctgtggtttc agggtccata
      961 aagcccacat ggataacatt acccctgaat tcgcatctag actgaactgg ccgataattg
     1021 cagacgaacg aagaggatgt ccaatatttt ttttaaggaa taaggatact tcaagactag
     1081 attcccccct gcattcccat cagaaccgta aaccttggcg ctttccttgg gaagtattca
     1141 agaagtgcct tgtccggttt ctgtggctca caaaccagcg cgcccgatat ggctttcttt
     1201 tcacttatga atgtaccagt acgggacaat tagaacgctc ctgtaacaat ctctttgcaa
     1261 atgtggggtt acattctaac catgtcacac tgctgacgaa attcaaagta aaaaaaaatg
     1321 ggaccacgtc ttgagaacga tagattttct ttattttaca ttgaacagtc gttgtctcag
     1381 cgcgctttat gttttcattc atacttcata ttataaaata acaaaagaag aatttcatat
     1441 tcacgcccaa gaaatcaggc tgctttccaa atgcaattga cacttcatta gccatcacac
     1501 aaaactcttt cttgctggag cttcttttaa aaaagacctc agtacaccaa acacgttacc
     1561 cgacctcgtt attttacgac aactatgata aaattctgaa gaaaaaataa aaaaattttc
     1621 atacttcttg cttttattta aaccattgaa tgatttcttt tgaacaaaac tacctgtttc
     1681 accaaaggaa atagaaagaa aaaatcaatt agaagaaaac aaaaaacaaa agatctatga
     1741 gtcgtttaga taaatccaag gttataaacg gcgcactaga actgcttaat ggcgttggta
     1801 tcgaaggtct gactacgagg aagttagcac agaaacttgg ggtcgagcag cccacgctgt
     1861 attggcacgt aaagaacaag agagcgctat tagatgccct accaatagag atgttagata
     1921 gacaccatac tcatttttgc ccgcttgagg gggaaagctg gcaagacttt ttaagaaata
     1981 atgcaaagag ttacaggtgc gctcttcttt cacataggga tggggcaaag gtacacttag
     2041 gtacgaggcc aaccgagaaa cagtatgaaa cactagaaaa ccaacttgct tttttatgcc
     2101 aacagggttt cagtcttgag aatgcacttt atgcccttag cgcagtcggg cactttacgt
     2161 tgggttgcgt cctagaggag caggaacatc aagtcgcaaa agaggaacgt gagacaccga
     2221 ctactgatag tatgcccccg ttattacgtc aggcaataga actatttgac cgtcaagggg
     2281 cggaaccggc ttttcttttt ggcctggagc taattatttg tgggctggag aagcaactga
     2341 agtgcgaaag tggtgggcca actgacgcgc tagatgattt cgatcttgat atgttacctg
     2401 cagatgcact agatgacttt gatttagaca tgttgcctgg aggatcctaa ctcgaggcga
     2461 atttcttatg atttatgatt tttattatta aataagttat aaaaaaaata agtgtataca
     2521 aattttaaag tgactcttag gttttaaaac gaaaattctt attcttgagt aactctttcc
     2581 tgtaggtcag gttgctttct caggtatagc atgaggtcgc tcttattgac cacacctcta
     2641 ccggcatgcc gagcaaatgc ctgcaaatcg ctccccattt cgctgccaaa ccagatgtca
     2701 acacagctac aacgggtagt ccatcgttgt aggatactcc ctatcagtga tagagaaatc
     2761 tatgcggcat ccctatcagt gatagagatc ggtaacggag tccctatcag tgatagagaa
     2821 gtggtgttca gttccctatc agtgatagag aagtttgaca ctatccctat cagtgataga
     2881 gagaacagca atgactccct atcagtgata gagatcaaat ggcgtatccc tatcagtgat
     2941 agagataatt aacttgtaat attctaatca attgacaata ttatttaagg acctattgtt
     3001 ttttccaata ggtggttagc aatcgtctta ctttctaact tttcttacct tttacatttc
     3061 agcaatatat atatatatat ttcaaggata taccattcta agatctgttt aaacgagctc
     3121 gctagcctcg agtctagagt cgacctgcag gtaactcgag agtgctttta actaagaatt
     3181 attagtcttt tctgcttatt ttttcatcat agtttagaac actttatatt aacgaatagt
     3241 ttatgaatct atttaggttt aaaaattgat acagttttat aagttacttt ttcaaagact
     3301 cgtgctgtct attgcataat gcactggaag gggaaaaaaa aggtgcacac gcgtggcttt
     3361 ttcttgaatt tgcagtttga aaaatgctga gcagttacag agatgttacg aaccactagt
     3421 gcactgcagt acacggtttc cttgaaattt ttttgattcg gtaatctccg aacagaagga
     3481 agaacgaagg aaggagcaca gacttagatt ggtatatata cgcatatgta gtgttgaaga
     3541 aacatgaaat tgcccagtat tcttaaccca actgcacaga acaaaaacgt gcaggaaacg
     3601 aagataaatc atgtcgaaag ctacatataa ggaacgtgct gctactcatc ctagtcctgt
     3661 tgctgccaag ctatttaata tcatgcacga aaagcaaaca aacttgtgtg cttcattgga
     3721 tgttcgtacc accaaggaat tactggagtt agttgaagca ttaggtccca aaatttgttt
     3781 actaaaaaca catgtggata tcttgactga tttttccatg gagggcacag ttaagccgct
     3841 aaaggcatta tccgccaagt acaatttttt actcttcgag gacagaaaat ttgctgacat
     3901 tggtaataca gtcaaattgc agtactctgc gggtgtatac agaatagcag aatgggcaga
     3961 cattacgaat gcacacggtg tggtgggccc aggtattgtt agcggtttga agcaggcggc
     4021 agaagaagta acaaaggaac ctagaggcct tttgatgtta gcagaattgt catgcaaggg
     4081 ctccctatct actggagaat atactaaggg tactgttgac attgcgaaga gcgacaaaga
     4141 ttttgttatc ggctttattg ctcaaagaga catgggtgga agagatgaag gttacgattg
     4201 gttgattatg acacccggtg tgggtttaga tgacaaggga gatgcattgg gtcaacagta
     4261 tagaaccgtg gatgatgtgg tttctacagg atctgacatt attattgttg gaagaggact
     4321 atttgcaaag ggaagggatg ctaaggtaga gggtgaacgt tacagaaaag caggctggga
     4381 agcatatttg agaagatgcg gccagcaaaa ctaaaaaact gtattataag taaatgcatg
     4441 tatactaaac tcacaaatta gagcttcaat ttaattatat cagttattac ccgagtagag
     4501 cacttgaatc cactgccccg ggaatctcgg tcgtaatgat ttctataatg acgaaaaaaa
     4561 aaaaattgga aagaaaaagc ttcatggcct ttataaaaag gaactatcca atacctcgcc
     4621 agaaccaagt aacagtattt tacggggcac aaatcaagaa caataagaca ggactgtaaa
     4681 gatggacgca ttgaactcca aagaacaaca agagttccaa aaagtagtgg aacaaaagca
     4741 aatgaaggat ttcatgcgtt tgtactctaa tctggtagaa agatgtttca cagactgtgt
     4801 caatgacttc acaacatcaa agctaaccaa taaggaacaa acatgcatca tgaagtgctc
     4861 agaaaagttc ttgaagcata gcgaacgtgt agggcagcgt ttccaagaac aaaacgctgc
     4921 cttgggacaa ggcttgggcc gataaggtgt actggcgtat atatatctaa ttatgtatct
     4981 ctggtgtagc ccatttttag catgtaaata taaagaccga gcggccgcgt gttacaacca
     5041 attaaccaat tctgattaga aaaactcatc gagcatcaaa tgaaactgca atttattcat
     5101 atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc
     5161 accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc
     5221 aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc
     5281 accatgagtg acgactgaat ccggtgagaa tggcaaaagc ttatgcattt ctttccagac
     5341 ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt
     5401 attcattcgt gattgcgcct gagcgaggcg aaatacgcga tcgctgttaa aaggacaatt
     5461 acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc
     5521 acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt
     5581 gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa
     5641 ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt
     5701 gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc
     5761 acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt
     5821 ggaatttaat cgcggcctgg agcaagacgt ttcccgttga atatggctca taacacccct
     5881 tgtattactg tttatgtaag cagacagttt tattgttcat gatgatatat ttttatcttg
     5941 tgcaatgtaa catcagagat tttgagacac aacgtggctt tgttgaataa atcgaacttt
     6001 tgctgagttg aaggatcagt catgaccaaa atcccttaac gtgagttttc gttccactga
     6061 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6121 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6181 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6241 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6301 tacctcgctc tgctaatcct gttaccagtg gct
//
