LOCUS       sequence-468716-        7397 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo-YTK URA multigene plasmid with MCS; NS3 systems; 
            pTDH3-LexA-NLS-NS3-V1-VP48-2xOaf1-tADH1; lexAO6-pLEU2m-MCS-tENO2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7397)
  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 7397)
  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: pAL007
FEATURES             Location/Qualifiers
     source          1..7397
                     /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(1663..2478)
                     /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(1663..2478)
                     /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     2386..2405
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2470..2489
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2703..3291
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3192..3211
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3984..4650
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     4520..4543
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     CDS             join(4657..4908,4909..5262)
                     /codon_start=1
                     /gene="lexA"
                     /product="E. coli DNA-binding protein that represses genes 
                     involved in the SOS response to DNA damage"
                     /label=LexA
                     /note="The N-terminal DNA-binding domain recognizes the 
                     16-bp palindromic sequence 5'-CTGTATATATATACAG-3'."
                     /translation="MKALTARQQEVFDLIRDHISQTGMPPTRAEIAQRLGFRSPNAAEE
                     HLKALARKGVIEIVSGASRGIRLLQEEEEGLPLVGRVAAGEPLLAQQHIEGHYQVDPSL
                     FKPNADFLLRVSGMSMKDIGIMDGDLLAVHKTQDVRNGQVVVARIDDEVTVKRLKKQGN
                     KVELLPENSEFKPIVVDLRQQSFTIEGLAVGVIRNGDWL"
     CDS             5278..5298
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6160..6186
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     terminator      6497..6684
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(6552..6576)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
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 aattggaaag aaaaagcttc atggccttta
     1201 taaaaaggaa ctatccaata cctcgccaga accaagtaac agtattttac ggggcacaaa
     1261 tcaagaacaa taagacagga ctgtaaagat ggacgcattg aactccaaag aacaacaaga
     1321 gttccaaaaa gtagtggaac aaaagcaaat gaaggatttc atgcgtttgt actctaatct
     1381 ggtagaaaga tgtttcacag actgtgtcaa tgacttcaca acatcaaagc taaccaataa
     1441 ggaacaaaca tgcatcatga agtgctcaga aaagttcttg aagcatagcg aacgtgtagg
     1501 gcagcgtttc caagaacaaa acgctgcctt gggacaaggc ttgggccgat aaggtgtact
     1561 ggcgtatata tatctaatta tgtatctctg gtgtagccca tttttagcat gtaaatataa
     1621 agaccgagcg gccgcgtgtt acaaccaatt aaccaattct gattagaaaa actcatcgag
     1681 catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag
     1741 ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg
     1801 gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc
     1861 aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg
     1921 caaaagctta tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc
     1981 aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgaggcgaaa
     2041 tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa
     2101 cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa
     2161 tgctgttttc ccggggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa
     2221 atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc
     2281 tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg
     2341 cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt
     2401 atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctggagc aagacgtttc
     2461 ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag acagttttat
     2521 tgttcatgat gatatatttt tatcttgtgc aatgtaacat cagagatttt gagacacaac
     2581 gtggctttgt tgaataaatc gaacttttgc tgagttgaag gatcagtcat gaccaaaatc
     2641 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     2701 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     2761 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     2821 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     2881 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     2941 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     3001 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     3061 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     3121 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     3181 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     3241 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     3301 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     3361 gcgttatccc ctgattctgt ggataaccgt gcggccgcca atgctaaatt cgagtgaaac
     3421 acaggaagat cagaaaatcc tcatttcatc catattaaca ataatttcaa atgtttattt
     3481 gcattatttg aaactaggca agacaagcaa cgaaacgttt ttgaaaattt tgagtatttt
     3541 caataaattt gtagaggact cagatattga aaaaaagcta cagcaattaa tacttgataa
     3601 gaagagtatt gagaagggca acggttcatc atctcatgga tctgcacatg aacaaacacc
     3661 agagtcaaac gacgttgaaa ttgaggctac tgcgccaatt gatgacaata cagacgatga
     3721 taacaaaccg aagttatctg atgtagaaaa ggattaaaga tgctaagaga tagtgatgat
     3781 atttcataaa taatgtaatt ctatatatgt taattacctt ttttgcgagg catatttatg
     3841 gtgaaggata agttttgacc atcaaagaag gttaatgtgg ctgtggtttc agggtccata
     3901 aagcccacat ggataacatt acccctgaat tcgcatctag actgaactgg ccgataattg
     3961 cagacgaacg cagttcgagt ttatcattat caatactgcc atttcaaaga atacgtaaat
     4021 aattaatagt agtgattttc ctaactttat ttagtcaaaa aattagcctt ttaattctgc
     4081 tgtaacccgt acatgcccaa aatagggggc gggttacaca gaatatataa catcgtaggt
     4141 gtctgggtga acagtttatt cctggcatcc actaaatata atggagcccg ctttttaagc
     4201 tggcatccag aaaaaaaaag aatcccagca ccaaaatatt gttttcttca ccaaccatca
     4261 gttcataggt ccattctctt agcgcaacta cagagaacag gggcacaaac aggcaaaaaa
     4321 cgggcacaac ctcaatggag tgatgcaacc tgcctggagt aaatgatgac acaaggcaat
     4381 tgacccacgc atgtatctat ctcattttct tacaccttct attaccttct gctctctctg
     4441 atttggaaaa agctgaaaaa aaaggttgaa accagttccc tgaaattatt cccctacttg
     4501 actaataagt atataaagac ggtaggtatt gattgtaatt ctgtaaatct atttcttaaa
     4561 cttcttaaat tctactttta tagttagtct tttttttagt tttaaaacac caagaactta
     4621 gtttcgaata aacacacata aacaaacaaa agatctatga aggctttaac tgcaagacaa
     4681 caggaagttt ttgatttgat aagagatcat atatctcaaa ctggaatgcc accaacaaga
     4741 gctgaaatag ctcaaagatt aggatttaga tctccaaacg ctgcagaaga acatttgaaa
     4801 gctttggcaa ggaaaggtgt catcgaaatt gtctctggtg cttcaagagg tattagacta
     4861 ttacaagaag aggaagaagg attgccattg gttggtagag ttgctgctgg tgaaccatta
     4921 ttagcacagc aacatattga gggtcattat caagtagatc catctttatt caaaccaaat
     4981 gctgattttt tgttgagagt aagtggtatg tctatgaaag atattggtat catggatggt
     5041 gacttattgg ctgttcataa aactcaagat gttagaaacg gtcaagttgt cgttgccaga
     5101 attgatgatg aagtcactgt taaaagattg aagaaacaag gtaataaagt ggaattgtta
     5161 ccagaaaatt ctgaatttaa gccaattgtt gttgatttaa ggcaacaatc ttttactatt
     5221 gaaggtttag ctgtcggtgt tatcagaaat ggtgattggt taggttcagg ttctcctcca
     5281 aaaaaaaaga gaaaagttgg atctggttct ggggcgagcg cgggcatggc gtcaatgact
     5341 gggggccagc aaatggggag tacggaggat gtagtttgtt gtcattcaat atacggcaag
     5401 aagaagggtg acatcgacac gtacagatac ataggctcta gtgggacagg ctgcgtagta
     5461 atcgtgggca gaattgtcct aagcggatcc ggaaccagtg cgcctatcac cgcctacgca
     5521 caacaaacca ggggactatt gggctgtatc ataacaagcc taactggaag agacaaaaat
     5581 caagtcgagg gcgaggttca gatcgtatct acggcaactc agacctttct tgccacgtgc
     5641 ataaacggcg tgtgctgggc cgtatatcat ggggctggca caagaacgat agcgagtcct
     5701 aagggtcctg ttattcaaat gtataccaac gtagaccagg atctagtcgg ctggcccgcg
     5761 ccccaaggct ctaggagtct taccccttgc acgtgcgggt cctcagactt atacctggta
     5821 acgaggcacg cagatgttat acctgtgcgt agaaggggtg acagtagagg atctctttta
     5881 tccccccgtc ccattagcta cttaaagggt agctctgggg ggccgttgtt atgtccggca
     5941 ggacatgccg tcgggttatt cagagcggct gtttgtacga ggggtgttgc aaaagcagta
     6001 gattttatac cggttgagaa cttagagaca acgatgaggt ccccggtttt tacagacaac
     6061 tcatctccac ctgctgttac actgacccac cccatcacca aaattgacag ggaggttctc
     6121 taccaggaat ttgacgaaat ggaggagtgc tcacaacact atccatacga cgtaccagat
     6181 tatgctggat ccccggcgga tgctctagac gatttcgatc tggatatgct accagccgat
     6241 gcacttgacg atttcgactt agacatgctt ccggccgacg ctctagatga cttcgactta
     6301 gatatgctcg gttccggcgc gaacaataca ccatttccgg gatattttgg gggccttgat
     6361 ttattcgact acgacttcct tttcggcaac gactttgctg gctcaggtgc gaataatact
     6421 ccttttccag gttattttgg aggtttggat ttatttgatt atgacttttt gtttggcaat
     6481 gactttgctt aatggcgcga atttcttatg atttatgatt tttattatta aataagttat
     6541 aaaaaaaata agtgtataca aattttaaag tgactcttag gttttaaaac gaaaattctt
     6601 attcttgagt aactctttcc tgtaggtcag gttgctttct caggtatagc atgaggtcgc
     6661 tcttattgac cacacctcta ccggcatgcc gagcaaatgc ctgcaaatcg ctccccattt
     6721 cgctgccaaa ccagatgtca acacagctac aacgggtagt ccatcgttgt aggatactgt
     6781 atatacaccc agtagagtag gtgactactg tatgagcata cagtagtgac aaccacttac
     6841 tgtatataaa tacagtagtg gtcatcgata ctgtatataa aaccagtagg caatcgtttg
     6901 tactgtatgt acatacagta tacctcgcaa ctactgtata taaacacagt aattaacttg
     6961 taatattcta atcaattgac aatattattt aaggacctat tgttttttcc aataggtggt
     7021 tagcaatcgt cttactttct aacttttctt accttttaca tttcagcaat atatatatat
     7081 atatttcaag gatataccat tctaagatct gtttaaacga gctcgctagc ctcgagtcta
     7141 gagtcgacct gcaggtaact cgagagtgct tttaactaag aattattagt cttttctgct
     7201 tattttttca tcatagttta gaacacttta tattaacgaa tagtttatga atctatttag
     7261 gtttaaaaat tgatacagtt ttataagtta ctttttcaaa gactcgtgct gtctattgca
     7321 taatgcactg gaaggggaaa aaaaaggtgc acacgcgtgg ctttttcttg aatttgcagt
     7381 ttgaaaaatg ctgagca
//
