LOCUS       sequence-468718-        8966 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo-YTK URA multigene plasmid with MCS; ABA systems; 
            pTDH3-PhlF-NES-PYL1-tADH1; pTDH3-NLS-VP48-2xOaf1-ABI-tENO1; 
            phlfO7-pLEU2m-MCS-tENO2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8966)
  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 8966)
  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: pAL005
FEATURES             Location/Qualifiers
     source          1..8966
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             218..1021
                     /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(1664..2479)
                     /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(1664..2479)
                     /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     2387..2406
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2471..2490
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2704..3292
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3193..3212
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3985..4651
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     CDS             5267..5293
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     terminator      5861..6048
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(5916..5940)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        6132..6798
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     CDS             6808..6828
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 gttacagaga tgttacgaac cactagtgca ctgcagtaca cggtttcctt gaaatttttt
       61 tgattcggta atctccgaac agaaggaaga acgaaggaag gagcacagac ttagattggt
      121 atatatacgc atatgtagtg ttgaagaaac atgaaattgc ccagtattct taacccaact
      181 gcacagaaca aaaacgtgca ggaaacgaag ataaatcatg tcgaaagcta catataagga
      241 acgtgctgct actcatccta gtcctgttgc tgccaagcta tttaatatca tgcacgaaaa
      301 gcaaacaaac ttgtgtgctt cattggatgt tcgtaccacc aaggaattac tggagttagt
      361 tgaagcatta ggtcccaaaa tttgtttact aaaaacacat gtggatatct tgactgattt
      421 ttccatggag ggcacagtta agccgctaaa ggcattatcc gccaagtaca attttttact
      481 cttcgaggac agaaaatttg ctgacattgg taatacagtc aaattgcagt actctgcggg
      541 tgtatacaga atagcagaat gggcagacat tacgaatgca cacggtgtgg tgggcccagg
      601 tattgttagc ggtttgaagc aggcggcaga agaagtaaca aaggaaccta gaggcctttt
      661 gatgttagca gaattgtcat gcaagggctc cctatctact ggagaatata ctaagggtac
      721 tgttgacatt gcgaagagcg acaaagattt tgttatcggc tttattgctc aaagagacat
      781 gggtggaaga gatgaaggtt acgattggtt gattatgaca cccggtgtgg gtttagatga
      841 caagggagat gcattgggtc aacagtatag aaccgtggat gatgtggttt ctacaggatc
      901 tgacattatt attgttggaa gaggactatt tgcaaaggga agggatgcta aggtagaggg
      961 tgaacgttac agaaaagcag gctgggaagc atatttgaga agatgcggcc agcaaaacta
     1021 aaaaactgta ttataagtaa atgcatgtat actaaactca caaattagag cttcaattta
     1081 attatatcag ttattacccg agtagagcac ttgaatccac tgccccggga atctcggtcg
     1141 taatgatttc tataatgacg aaaaaaaaaa aaattggaaa gaaaaagctt catggccttt
     1201 ataaaaagga actatccaat acctcgccag aaccaagtaa cagtatttta cggggcacaa
     1261 atcaagaaca ataagacagg actgtaaaga tggacgcatt gaactccaaa gaacaacaag
     1321 agttccaaaa agtagtggaa caaaagcaaa tgaaggattt catgcgtttg tactctaatc
     1381 tggtagaaag atgtttcaca gactgtgtca atgacttcac aacatcaaag ctaaccaata
     1441 aggaacaaac atgcatcatg aagtgctcag aaaagttctt gaagcatagc gaacgtgtag
     1501 ggcagcgttt ccaagaacaa aacgctgcct tgggacaagg cttgggccga taaggtgtac
     1561 tggcgtatat atatctaatt atgtatctct ggtgtagccc atttttagca tgtaaatata
     1621 aagaccgagc ggccgcgtgt tacaaccaat taaccaattc tgattagaaa aactcatcga
     1681 gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa
     1741 gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct
     1801 ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt
     1861 caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg
     1921 gcaaaagctt atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat
     1981 caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgaggcgaa
     2041 atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga
     2101 acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga
     2161 atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa
     2221 aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat
     2281 ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg
     2341 gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt
     2401 tatacccata taaatcagca tccatgttgg aatttaatcg cggcctggag caagacgttt
     2461 cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta
     2521 ttgttcatga tgatatattt ttatcttgtg caatgtaaca tcagagattt tgagacacaa
     2581 cgtggctttg ttgaataaat cgaacttttg ctgagttgaa ggatcagtca tgaccaaaat
     2641 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     2701 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     2761 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     2821 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     2881 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     2941 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     3001 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     3061 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     3121 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     3181 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     3241 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     3301 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     3361 tgcgttatcc cctgattctg tggataaccg tgcggccgcc aatgctaaat tcgagtgaaa
     3421 cacaggaaga tcagaaaatc ctcatttcat ccatattaac aataatttca aatgtttatt
     3481 tgcattattt gaaactaggc aagacaagca acgaaacgtt tttgaaaatt ttgagtattt
     3541 tcaataaatt tgtagaggac tcagatattg aaaaaaagct acagcaatta atacttgata
     3601 agaagagtat tgagaagggc aacggttcat catctcatgg atctgcacat gaacaaacac
     3661 cagagtcaaa cgacgttgaa attgaggcta ctgcgccaat tgatgacaat acagacgatg
     3721 ataacaaacc gaagttatct gatgtagaaa aggattaaag atgctaagag atagtgatga
     3781 tatttcataa ataatgtaat tctatatatg ttaattacct tttttgcgag gcatatttat
     3841 ggtgaaggat aagttttgac catcaaagaa ggttaatgtg gctgtggttt cagggtccat
     3901 aaagcccaca tggataacat tacccctgaa ttcgcatcta gactgaactg gccgataatt
     3961 gcagacgaac gcagttcgag tttatcatta tcaatactgc catttcaaag aatacgtaaa
     4021 taattaatag tagtgatttt cctaacttta tttagtcaaa aaattagcct tttaattctg
     4081 ctgtaacccg tacatgccca aaataggggg cgggttacac agaatatata acatcgtagg
     4141 tgtctgggtg aacagtttat tcctggcatc cactaaatat aatggagccc gctttttaag
     4201 ctggcatcca gaaaaaaaaa gaatcccagc accaaaatat tgttttcttc accaaccatc
     4261 agttcatagg tccattctct tagcgcaact acagagaaca ggggcacaaa caggcaaaaa
     4321 acgggcacaa cctcaatgga gtgatgcaac ctgcctggag taaatgatga cacaaggcaa
     4381 ttgacccacg catgtatcta tctcattttc ttacaccttc tattaccttc tgctctctct
     4441 gatttggaaa aagctgaaaa aaaaggttga aaccagttcc ctgaaattat tcccctactt
     4501 gactaataag tatataaaga cggtaggtat tgattgtaat tctgtaaatc tatttcttaa
     4561 acttcttaaa ttctactttt atagttagtc ttttttttag ttttaaaaca ccaagaactt
     4621 agtttcgaat aaacacacat aaacaaacaa aagatctatg gcacgtacgc cctcacgttc
     4681 cagcataggt agccttagat caccacatac gcacaaagcc atcctgactt caactattga
     4741 aatactaaaa gaatgcgggt attctggact aagtatcgaa tcagtggcta gacgtgccgg
     4801 ggccagcaaa cccacaattt accgttggtg gaccaacaaa gccgctctaa tagccgaagt
     4861 gtacgaaaac gagtctgaac aggtccgtaa gttccctgat ttaggctcct ttaaggcgga
     4921 tttggatttc ttactgagaa acctatggaa agtatggagg gagacaattt gcggagaagc
     4981 atttaggtgc gtcattgctg aagcccaact tgatcccgcg acgcttacac aactaaaaga
     5041 tcagtttatg gagagaaggc gtgaaatgcc aaagaaatta gtagaaaacg ccatatcaaa
     5101 cggggagcta cccaaagaca ccaacagaga actgctatta gatatgatct ttggtttttg
     5161 ctggtacaga ctgttgactg aacaacttac tgtggagcaa gatatagagg agttcacctt
     5221 tctgttgata aacggagttt gcccagggac acaacgtgga agcggactgc ctccgttaga
     5281 aagactaacg ctgggttctg gtggaggtgc tccaacacaa gatgagttca cacaactatc
     5341 acagagtatc gcagaattcc atacctacca actaggaaat ggaagatgtt cttcactgct
     5401 tgcgcaaaga atccatgccc cgccggagac tgtgtggagt gtagttagga ggttcgacag
     5461 gccgcagata tacaagcact ttatcaaatc ctgcaatgtt agcgaagatt tcgagatgcg
     5521 tgttggatgc accagggacg taaatgttat atctggatta cctgcgaata cgtcaagaga
     5581 aaggttggac ttactagacg acgacagacg tgttaccgga ttttccataa cgggtggtga
     5641 acatcgtcta cgtaattata agagcgtgac tacagtccac cgttttgaaa aggaggaaga
     5701 ggaggagaga atatggaccg tggttctgga atcctacgtt gttgatgtac ccgaggggaa
     5761 tagcgaggag gatacacgtc tgttcgcgga taccgttatc agattgaatc ttcaaaaact
     5821 tgcttccatt acagaggcca tgaacggatc ctaactcgag gcgaatttct tatgatttat
     5881 gatttttatt attaaataag ttataaaaaa aataagtgta tacaaatttt aaagtgactc
     5941 ttaggtttta aaacgaaaat tcttattctt gagtaactct ttcctgtagg tcaggttgct
     6001 ttctcaggta tagcatgagg tcgctcttat tgaccacacc tctaccggca tgccgagcaa
     6061 atgcctgcaa atcgctcccc atttcgctgc caaaccagat gtcaacacag ctacaacgca
     6121 gttcgagttt atcattatca atactgccat ttcaaagaat acgtaaataa ttaatagtag
     6181 tgattttcct aactttattt agtcaaaaaa ttagcctttt aattctgctg taacccgtac
     6241 atgcccaaaa tagggggcgg gttacacaga atatataaca tcgtaggtgt ctgggtgaac
     6301 agtttattcc tggcatccac taaatataat ggagcccgct ttttaagctg gcatccagaa
     6361 aaaaaaagaa tcccagcacc aaaatattgt tttcttcacc aaccatcagt tcataggtcc
     6421 attctcttag cgcaactaca gagaacaggg gcacaaacag gcaaaaaacg ggcacaacct
     6481 caatggagtg atgcaacctg cctggagtaa atgatgacac aaggcaattg acccacgcat
     6541 gtatctatct cattttctta caccttctat taccttctgc tctctctgat ttggaaaaag
     6601 ctgaaaaaaa aggttgaaac cagttccctg aaattattcc cctacttgac taataagtat
     6661 ataaagacgg taggtattga ttgtaattct gtaaatctat ttcttaaact tcttaaattc
     6721 tacttttata gttagtcttt tttttagttt taaaacacca agaacttagt ttcgaataaa
     6781 cacacataaa caaacaaaag atctatgccc aagaagaagc gtaaagtgcc ggcggatgct
     6841 ctagacgatt tcgatctgga tatgctacca gccgatgcac ttgacgattt cgacttagac
     6901 atgcttccgg ccgacgctct agatgacttc gacttagata tgctcggttc cggcgcgaac
     6961 aatacaccat ttccgggata ttttgggggc cttgatttat tcgactacga cttccttttc
     7021 ggcaacgact ttgctggctc aggtgcgaat aatactcctt ttccaggtta ttttggaggt
     7081 ttggatttat ttgattatga ctttttgttt ggcaatgact ttgctggttc tacgagagtt
     7141 cctttgtatg gcttcacctc aatttgcggt agaagacccg agatggaagc cgccgtatcg
     7201 acgataccac gttttttaca atcctcctcc ggctcaatgt tagacggacg ttttgatccc
     7261 cagagtgcag ctcatttctt tggtgtttat gatggtcacg ggggtagcca ggtagccaac
     7321 tattgccgtg aaaggatgca tttagcgtta gctgaggaaa tcgccaaaga aaagcctatg
     7381 ttgtgtgatg gagacacctg gttggagaag tggaaaaagg cgttattcaa cagcttccta
     7441 agggtggata gtgagataga gtctgttgct ccggagactg ttggctcaac atccgtggtg
     7501 gccgtggtct tcccgagcca tatttttgtg gctaattgtg gtgactctcg tgcggtcttg
     7561 tgcaggggga agactgcatt acctttaagc gtcgatcata agccggacag ggaggacgaa
     7621 gcagcgagga tagaggccgc cggtggaaag gtaatacaat ggaatggtgc tagggtattc
     7681 ggcgtactgg caatgagtag atccattggt gatagatact tgaagccgag cataatccct
     7741 gaccctgaag taacagcggt taaaagggtg aaggaagacg attgccttat cttggcttct
     7801 gacggagtgt gggatgtcat gactgacgag gaagcctgcg aaatggcaag gaaaagaata
     7861 ctactttggc ataagaaaaa tgcggtagct ggggacgcaa gtctgttagc agatgaaaga
     7921 agaaaagaag gcaaggaccc cgccgcaatg tcagccgcgg agtacctgtc taaacttgct
     7981 attcagaggg ggagtaaaga taacatatcc gtggtcgtgg ttgatctaaa gggatcctaa
     8041 ctcgagagct tttgattaag ccttctagtc caaaaaacac gtttttttgt catttatttc
     8101 attttcttag aatagtttag tttattcatt ttatagtcac gaatgtttta tgattctata
     8161 tagggttgca aacaagcatt tttcatttta tgttaaaaca atttcaggtt taccttttat
     8221 tctgcttgtg gtgacgcgtg tatccgcccg ctcttttggt cacccatgta tgctggatgc
     8281 acacactggc ttaagatgac aacgggatga tacgaaacgt accgtatcgt taaggtgaat
     8341 gatacgaaac gtaccgtatc gttaaggtca atgatacgaa acgtaccgta tcgttaaggt
     8401 caatgatacg aaacgtaccg tatcgttaag gtgtatgata cgaaacgtac cgtatcgtta
     8461 aggtagatga tacgaaacgt accgtatcgt taaggtcgat gatacgaaac gtaccgtatc
     8521 gttaaggtcg gatctcagct agctttgaca atattattta aggacctatt gttttttcca
     8581 ataggtggtt agcaatcgtc ttactttcta acttttctta ccttttacat ttcagcaata
     8641 tatatatata tatttcaagg atataccatt ctaagatctg tttaaacgag ctcgctagcc
     8701 tcgagtctag agtcgacctg caggtaactc gagagtgctt ttaactaaga attattagtc
     8761 ttttctgctt attttttcat catagtttag aacactttat attaacgaat agtttatgaa
     8821 tctatttagg tttaaaaatt gatacagttt tataagttac tttttcaaag actcgtgctg
     8881 tctattgcat aatgcactgg aaggggaaaa aaaaggtgca cacgcgtggc tttttcttga
     8941 atttgcagtt tgaaaaatgc tgagca
//
