LOCUS       sequence-243265-        8370 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dual inducible expression of Venus-degronSwitchA and KeyA-BFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8370)
  AUTHORS   Langan RA, Boyken SE, Ng AH, Samson JA, Dods G, Westbrook AM, Nguyen
            TH, Lajoie MJ, Chen Z, Berger S, Mulligan VK, Dueber JE, Novak WRP, 
            El-Samad H, Baker D
  TITLE     De novo design of bioactive protein switches.
  JOURNAL   Nature. 2019 Jul 24. pii: 10.1038/s41586-019-1432-8. doi: 
            10.1038/s41586-019-1432-8.
  PUBMED    31341284
REFERENCE   2  (bases 1 to 8370)
  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"; journalName: "Nature. 
            2019 Jul 24. pii: 10.1038/s41586-019-1432-8. doi: 
            10.1038/s41586-019-1432-8."
COMMENT     Sequence Label: pAN1572
FEATURES             Location/Qualifiers
     source          1..8370
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        106..547
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    106..223
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     CDS             565..1278
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEGGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             4643..5446
                     /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(6088..6903)
                     /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(6088..6903)
                     /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     6811..6830
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6895..6914
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      7128..7716
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7617..7636
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 actggccgat aattgcagac gaacgcccca ttatcttagc ctaaaaaaac cttctctttg
       61 gaactttcag taatacgctt aactgctcat tgctatattg aagtacggat tagaagccgc
      121 cgagcgggcg acagccctcc gacggatgac tctcctccgt gcgtcctcgt catcaccggt
      181 cgcgttcctg aaacgcagat gtgcctcgcg ccgcactgct ccgaacaata aagattctac
      241 aatactagct tttatggtta tgaagaggaa aaattggcag taacctggcc ccacaaacct
      301 tcaaattaac gaatcaaatt aacaaccata ggatgataat gcgattagtt ttttagcctt
      361 atttctgggg taattaatca gcgaagcgat gatttttgat ctattaacag atatataaat
      421 ggaaaagctg cataaccact ttaactaata ctttcaacat tttcagtttg tattacttct
      481 tattcaaatg tcataaaagt atcaacaaaa aattgttaat atacctctat actttaacgt
      541 caaggagaaa aaactataag atctatgtct aaaggtgaag aattattcac tggtgttgtc
      601 ccaattttgg ttgaattaga tggtgatgtt aatggtcaca aattttctgt ctccggtgaa
      661 ggtgaaggtg atgctactta cggtaaattg accttaaaat tgatttgtac tactggtaaa
      721 ttgccagttc catggccaac cttagtcact actttaggtt atggtttgca atgttttgct
      781 agatacccag atcatatgaa acaacatgac tttttcaagt ctgccatgcc agaaggttat
      841 gttcaagaaa gaactatttt tttcaaagat gacggtaact acaagaccag agctgaagtc
      901 aagtttgaag gtgatacctt agttaataga atcgaattaa aaggtattga ttttaaagaa
      961 ggtggtaaca ttttaggtca caaattggaa tacaactata actctcacaa tgtttacatc
     1021 actgctgaca aacaaaagaa tggtatcaaa gctaacttca aaattagaca caacattgaa
     1081 gatggtggtg ttcaattagc tgaccattat caacaaaata ctccaattgg tgatggtcca
     1141 gtcttgttac cagacaacca ttacttatcc tatcaatctg ccttatccaa agatccaaac
     1201 gaaaagagag atcacatggt cttgttagaa tttgttactg ctgctggtat tacccatggt
     1261 atggatgaat tgtacaaagg atccagtaag gaagcagtga cgaagctaca ggcccttaac
     1321 atcaaattgg cggaaaaatt gcttgaagct gtcacgaagt tgcaagcatt aaacataaaa
     1381 cttgccgaaa agctgttaga agccctggca cgtctgcaag agctgaacat tgccttggtt
     1441 tatttagccg tagaattaac ggaccctaag agaattgcgg acgagattaa aaaagttaaa
     1501 gacaagagca aggaaattgt cgagagagcg gaagaggaga tagctagggc ggcagccgag
     1561 tccaagaaaa ttttagacga agccgaggag gaaatcgccc gtgcagcagc agagtccaag
     1621 aagatccttg atgaaggatc cggaagtggt tcagacgcgg tagcggaatt acaggcatta
     1681 aatctgaagc tagccgagtt acttctggag gctgtggctg agcttcaagc gcttaacctg
     1741 aaactagcgg agctactttt ggaggctata gctaagctac aagagctgaa tattaagctt
     1801 gtcgagctat tgactaaact aactgaccct gctaccatca gggaggcgat cagaaaggtg
     1861 aaggaagact ccgagaggat agtggcggag gcagagcgtt tgatagccgc tgcgaaggct
     1921 gaaagtgaac gtattatacg tgaagctgag agattgatag ctgctgctaa agcggagtcc
     1981 gagaggataa ttagagaggg gtccggttca ggggaccctg acgtagcgag actgcaagaa
     2041 ctaaatatcg agctagctag agagctgcta agggatgtcg ctagactaca agagctaaac
     2101 atagaactgg cgagagaact tctaagggca gcggcagagt tacaagagct gaacataaag
     2161 ttagtcgagc tagcgagtga gttaaccgat cctgatgaag cacgtaaagc aatcgccagg
     2221 gtaaagagag agagtaaaag aatcgttgag gatcttatta tgagctgtgc ccaagagtct
     2281 gctgcaagcg agaagattag cagagaagcc gaaagactga taaggtaatg gcgccaattg
     2341 gtgcggcaat tgataataac gaaaatgtct tttaatgatc tgggtataat gaggaatttt
     2401 ccgaacgttt ttactttata tatatatata catgtaacat atattctata cgctatagag
     2461 aaaggaaatt tttcaattaa aaaaaaaata gagaaagagt ttcacttctt gattatcgct
     2521 aacactaatg gttgaagtac tgctacttta attttatgct gccaaaccag atgtcaacac
     2581 agctacaacg ttatattgaa ttttcaaaaa ttcttacttt ttttttggat ggacgcaaag
     2641 aagtttaata atcatattac atggcattac caccatatac atatccatat acatatccat
     2701 atctaatctt acttatatgt tgtggaaatg taaagagccc cattatctta gcctaaaaaa
     2761 accttctctt tggaactttc agtaatacgc ttaactgctc attgctatat tgaagtgtgg
     2821 ccgcgtgggc gtgcgtgggc gggcgtgggc gtgcgtgggc gggcgtgggc gtgcgtgggc
     2881 gtctagaccg tgcgtcctcg tcttcaccgg tcgcgttcct gaaacgcaga tgtgcctaat
     2941 gccgcactgc tccgaacaat aaagattcta caatactagc ttttatggtt atgaagagga
     3001 aaaattggca gtaacctggc cccacaaacc ttcaaattaa cgaatcaaat taacaaccat
     3061 aggatgataa tgcgattagt tttttagcct tatttctggg gtaattaatc agcgaagcga
     3121 tgatttttga tctattaaca gatatataaa tggaaaagct gcataaccac tttaactaat
     3181 actttcaaca ttttcagttt gtattacttc ttattcaaat gtcataaaag tatcaacaaa
     3241 aaattgttaa tatacctcta tactttaacg tcaaggagaa aaaactataa gatctatgga
     3301 ggctagaaaa gccatcgcta gagtaaagag agaatcaaag agaatcgtag aagatgctga
     3361 gaggttaata agagaagcag ccgccgcatc tgagaaaatt tcaagggagg ccgaaaggtt
     3421 gattagggag gcagctgcgg cttcagaaaa gataagtagg gagggttctg tttcaaaggg
     3481 tgaagaattg atcaaagaaa acatgcatat gaaattatat atggaaggta cagttgacaa
     3541 ccatcatttt aagtgtacat ctgaaggaga aggtaagcct tacgagggaa ctcaaactat
     3601 gagaataaaa gtcgtagaag gtggaccatt gccttttgct tttgatatct tagcaacatc
     3661 tttcttatat ggttctaaaa cctttattaa ccatactcaa ggtatccctg atttcttcaa
     3721 acaatctttc cctgaaggat ttacttggga gagggttaca acatatgagg atggtggtgt
     3781 tttgacagca acccaggata catcattgca ggatggatgt ttgatttata atgtaaagat
     3841 tagaggagta aattttactt ctaacggtcc tgtaatgcaa aagaagactt taggttggga
     3901 agctttcacc gaaacattat atccagcaga cggaggttta gagggaagaa acgatatggc
     3961 tttgaagtta gtcggtggtt cacacttaat agctaacgcc aagactactt acaggtctaa
     4021 gaaaccagca aagaatttga aaatgccagg agtttattat gttgactaca gattggaaag
     4081 gatcaaggag gccaataacg agacatatgt agagcagcat gaagtcgccg tcgccagata
     4141 ttgcgactta ccttctaagt tgggtcataa attgaatgga tcctaactcg agagcttttg
     4201 attaagcctt ctagtccaaa aaacacgttt ttttgtcatt tatttcattt tcttagaata
     4261 gtttagttta ttcattttat agtcacgaat gttttatgat tctatatagg gttgcaaaca
     4321 agcatttttc attttatgtt aaaacaattt caggtttacc ttttattctg cttgtggtga
     4381 cgcgtgtatc cgcccgctct tttggtcacc catgtatgct gagcagttac agagatgtta
     4441 cgaaccacta gtgcactgca gtacacggtt tccttgaaat ttttttgatt cggtaatctc
     4501 cgaacagaag gaagaacgaa ggaaggagca cagacttaga ttggtatata tacgcatatg
     4561 tagtgttgaa gaaacatgaa attgcccagt attcttaacc caactgcaca gaacaaaaac
     4621 gtgcaggaaa cgaagataaa tcatgtcgaa agctacatat aaggaacgtg ctgctactca
     4681 tcctagtcct gttgctgcca agctatttaa tatcatgcac gaaaagcaaa caaacttgtg
     4741 tgcttcattg gatgttcgta ccaccaagga attactggag ttagttgaag cattaggtcc
     4801 caaaatttgt ttactaaaaa cacatgtgga tatcttgact gatttttcca tggagggcac
     4861 agttaagccg ctaaaggcat tatccgccaa gtacaatttt ttactcttcg aagacagaaa
     4921 atttgctgac attggtaata cagtcaaatt gcagtactct gcgggtgtat acagaatagc
     4981 agaatgggca gacattacga atgcacacgg tgtggtgggc ccaggtattg ttagcggttt
     5041 gaagcaggcg gcagaagaag taacaaagga acctagaggc cttttgatgt tagcagaatt
     5101 gtcatgcaag ggctccctat ctactggaga atatactaag ggtactgttg acattgcgaa
     5161 gagcgacaaa gattttgtta tcggctttat tgctcaaaga gacatgggtg gaagagatga
     5221 aggttacgat tggttgatta tgacacccgg tgtgggttta gatgacaagg gagatgcatt
     5281 gggtcaacag tatagaaccg tggatgatgt ggtttctaca ggatctgaca ttattattgt
     5341 tggaagagga ctatttgcaa agggaaggga tgctaaggta gagggtgaac gttacagaaa
     5401 agcaggctgg gaagcatatt tgagaagatg cggccagcaa aactaaaaaa ctgtattata
     5461 agtaaatgca tgtatactaa actcacaaat tagagcttca atttaattat atcagttatt
     5521 acccgagtag agcacttgaa tccactgccc cgggaatctc ggtcgtaatg atttctataa
     5581 tgacgaaaaa aaaaaaattg gaaagaaaaa gcttcatggc ctttataaaa aggaactatc
     5641 caatacctcg ccagaaccaa gtaacagtat tttacggggc acaaatcaag aacaataaga
     5701 caggactgta aagatggacg cattgaactc caaagaacaa caagagttcc aaaaagtagt
     5761 ggaacaaaag caaatgaagg atttcatgcg tttgtactct aatctggtag aaagatgttt
     5821 cacagactgt gtcaatgact tcacaacatc aaagctaacc aataaggaac aaacatgcat
     5881 catgaagtgc tcagaaaagt tcttgaagca tagcgaacgt gtagggcagc gtttccaaga
     5941 acaaaacgct gccttgggac aaggcttggg ccgataaggt gtactggcgt atatatatct
     6001 aattatgtat ctctggtgta gcccattttt agcatgtaaa tataaagacc gagcggccgc
     6061 gtgttacaac caattaacca attctgatta gaaaaactca tcgagcatca aatgaaactg
     6121 caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga
     6181 aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat
     6241 tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc
     6301 aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa gcttatgcat
     6361 ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc
     6421 aaccaaaccg ttattcattc gtgattgcgc ctgagcgagg cgaaatacgc gatcgctgtt
     6481 aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc
     6541 aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg ttttcccggg
     6601 gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg
     6661 aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc
     6721 aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg
     6781 atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc
     6841 agcatccatg ttggaattta atcgcggcct ggagcaagac gtttcccgtt gaatatggct
     6901 cataacaccc cttgtattac tgtttatgta agcagacagt tttattgttc atgatgatat
     6961 atttttatct tgtgcaatgt aacatcagag attttgagac acaacgtggc tttgttgaat
     7021 aaatcgaact tttgctgagt tgaaggatca gtcatgacca aaatccctta acgtgagttt
     7081 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     7141 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     7201 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     7261 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     7321 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     7381 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     7441 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     7501 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     7561 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     7621 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     7681 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     7741 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     7801 tctgtggata accgtgcggc cgccaatgct aaattcgagt gaaacacagg aagatcagaa
     7861 aatcctcatt tcatccatat taacaataat ttcaaatgtt tatttgcatt atttgaaact
     7921 aggcaagaca agcaacgaaa cgtttttgaa aattttgagt attttcaata aatttgtaga
     7981 ggactcagat attgaaaaaa agctacagca attaatactt gataagaaga gtattgagaa
     8041 gggcaacggt tcatcatctc atggatctgc acatgaacaa acaccagagt caaacgacgt
     8101 tgaaattgag gctactgcgc caattgatga caatacagac gatgataaca aaccgaagtt
     8161 atctgatgta gaaaaggatt aaagatgcta agagatagtg atgatatttc ataaataatg
     8221 taattctata tatgttaatt accttttttg cgaggcatat ttatggtgaa ggataagttt
     8281 tgaccatcaa agaaggttaa tgtggctgtg gtttcagggt ccataaagcc cacatggata
     8341 acattacccc tgaattcgca tctagactga
//
