LOCUS       sequence-243263-        9112 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dual inducible expression of SynTF-mScarlet-degronSwitchA and 
            KeyA-BFP-NLS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9112)
  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 9112)
  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: pAN1735
FEATURES             Location/Qualifiers
     source          1..9112
                     /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             568..633
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             640..660
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4895..4915
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5385..6188
                     /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(6830..7645)
                     /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(6830..7645)
                     /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     7553..7572
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     7637..7656
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      7870..8458
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8359..8378
                     /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 atctatggat tataaggatc acgatggcga ctacaaggac
      601 cacgatattg actacaaaga cgatgacgat aaaatggctc ctaagaaaaa gcgcaaagtc
      661 ggtatccatg gcgttcccgt cgacggtgga ctgcaggccc caccaaccga tgtttcttta
      721 ggtgatgagt tacatttgga cggtgaggac gtagctatgg cacatgcaga tgccttagac
      781 gattttgact tggatatgtt aggagatgga gattcaccag gacctggttt cacaccacac
      841 gattcagctc catacggtgc tttggacatg gccgatttcg aatttgaaca gatgtttact
      901 gatgccttgg gtatagacga atacggtggt ggagctagac ccggggagcg ccccttccag
      961 tgtcgcattt gcatgcggaa cttttcgcgc caggacaggc ttgacaggca tacccgtact
     1021 cataccggtg aaaaaccgtt tcagtgtcgg atctgtatgc gaaatttctc ccagaaggag
     1081 cacttggcgg ggcatctacg tacgcacacc ggcgagaagc cattccaatg ccgaatatgc
     1141 atgcgcaact tcagtcgccg cgacaacctg aaccggcacc taaaaaccca cctgagagga
     1201 tctggttctg tatcaaaagg cgaggcggtc attaaagagt tcatgaggtt taaagtccat
     1261 atggaaggaa gcatgaatgg tcacgagttt gagatagagg gagagggtga gggaaggccg
     1321 tacgaaggga cgcaaactgc gaagttgaag gtaacgaaag gcggccctct gccattctct
     1381 tgggacatcc tttcaccgca atttatgtac ggctctaggg ccttcacaaa acaccccgcc
     1441 gatattccgg attactataa acaaagtttt cctgagggct tcaagtggga aagagtaatg
     1501 aacttcgagg atgggggagc tgtgacggtt actcaagaca ctagcctgga ggacggtact
     1561 cttatataca aggtaaaact aagagggacg aattttccgc cagacggccc cgtgatgcaa
     1621 aaaaaaacta tggggtggga agcatccacc gagaggctgt accccgaaga cggggtcctg
     1681 aagggtgata ttaaaatggc attaagatta aaagatggcg ggagatactt agctgacttc
     1741 aagactacat ataaagccaa aaaacctgta cagatgcctg gagcttacaa tgtggatcgt
     1801 aagctggata ttacatccca caacgaagat tataccgtgg tggagcagta cgagcgttca
     1861 gaggggcgtc actccaccgg gggtatggat gaattataca agagcggtct tagaagcagg
     1921 gcgcaagcgt caaattcagc ggtagatgga acagcaggtc caggtagtac tggcagcaga
     1981 ggatccagta aggaagcagt gacgaagcta caggccctta acatcaaatt ggcggaaaaa
     2041 ttgcttgaag ctgtcacgaa gttgcaagca ttaaacataa aacttgccga aaagctgtta
     2101 gaagccctgg cacgtctgca agagctgaac attgccttgg tttatttagc cgtagaatta
     2161 acggacccta agagaattgc ggacgagatt aaaaaagtta aagacaagag caaggaaatt
     2221 gtcgagagag cggaagagga gatagctagg gcggcagccg agtccaagaa aattttagac
     2281 gaagccgagg aggaaatcgc ccgtgcagca gcagagtcca agaagatcct tgatgaagga
     2341 tccggaagtg gttcagacgc ggtagcggaa ttacaggcat taaatctgaa gctagccgag
     2401 ttacttctgg aggctgtggc tgagcttcaa gcgcttaacc tgaaactagc ggagctactt
     2461 ttggaggcta tagctaagct acaagagctg aatattaagc ttgtcgagct attgactaaa
     2521 ctaactgacc ctgctaccat cagggaggcg atcagaaagg tgaaggaaga ctccgagagg
     2581 atagtggcgg aggcagagcg tttgatagcc gctgcgaagg ctgaaagtga acgtattata
     2641 cgtgaagctg agagattgat agctgctgct aaagcggagt ccgagaggat aattagagag
     2701 gggtccggtt caggggaccc tgacgtagcg agactgcaag aactaaatat cgagctagct
     2761 agagagctgc taagggatgt cgctagacta caagagctaa acatagaact ggcgagagaa
     2821 cttctaaggg cagcggcaga gttacaagag ctgaacataa agttagtcga gctagcgagt
     2881 gagttaaccg atcctgatga agcacgtaaa gcaatcgcca gggtaaagag agagagtaaa
     2941 agaatcgttg aggatcttat tatgagctgt gcccaagagt ctgctgcaag cgagaagatt
     3001 agcagagaag ccgaaagact gataaggtaa tggcgccaat tggtgcggca attgataata
     3061 acgaaaatgt cttttaatga tctgggtata atgaggaatt ttccgaacgt ttttacttta
     3121 tatatatata tacatgtaac atatattcta tacgctatag agaaaggaaa tttttcaatt
     3181 aaaaaaaaaa tagagaaaga gtttcacttc ttgattatcg ctaacactaa tggttgaagt
     3241 actgctactt taattttatg ctgccaaacc agatgtcaac acagctacaa cgttatattg
     3301 aattttcaaa aattcttact ttttttttgg atggacgcaa agaagtttaa taatcatatt
     3361 acatggcatt accaccatat acatatccat atacatatcc atatctaatc ttacttatat
     3421 gttgtggaaa tgtaaagagc cccattatct tagcctaaaa aaaccttctc tttggaactt
     3481 tcagtaatac gcttaactgc tcattgctat attgaagtgt ggccgcgtgg gcgtgcgtgg
     3541 gcgggcgtgg gcgtgcgtgg gcgggcgtgg gcgtgcgtgg gcgtctagac cgtgcgtcct
     3601 cgtcttcacc ggtcgcgttc ctgaaacgca gatgtgccta atgccgcact gctccgaaca
     3661 ataaagattc tacaatacta gcttttatgg ttatgaagag gaaaaattgg cagtaacctg
     3721 gccccacaaa ccttcaaatt aacgaatcaa attaacaacc ataggatgat aatgcgatta
     3781 gttttttagc cttatttctg gggtaattaa tcagcgaagc gatgattttt gatctattaa
     3841 cagatatata aatggaaaag ctgcataacc actttaacta atactttcaa cattttcagt
     3901 ttgtattact tcttattcaa atgtcataaa agtatcaaca aaaaattgtt aatatacctc
     3961 tatactttaa cgtcaaggag aaaaaactat aagatctatg gaggctagaa aagccatcgc
     4021 tagagtaaag agagaatcaa agagaatcgt agaagatgct gagaggttaa taagagaagc
     4081 agccgccgca tctgagaaaa tttcaaggga ggccgaaagg ttgattaggg aggcagctgc
     4141 ggcttcagaa aagataagta gggagggttc tgtttcaaag ggtgaagaat tgatcaaaga
     4201 aaacatgcat atgaaattat atatggaagg tacagttgac aaccatcatt ttaagtgtac
     4261 atctgaagga gaaggtaagc cttacgaggg aactcaaact atgagaataa aagtcgtaga
     4321 aggtggacca ttgccttttg cttttgatat cttagcaaca tctttcttat atggttctaa
     4381 aacctttatt aaccatactc aaggtatccc tgatttcttc aaacaatctt tccctgaagg
     4441 atttacttgg gagagggtta caacatatga ggatggtggt gttttgacag caacccagga
     4501 tacatcattg caggatggat gtttgattta taatgtaaag attagaggag taaattttac
     4561 ttctaacggt cctgtaatgc aaaagaagac tttaggttgg gaagctttca ccgaaacatt
     4621 atatccagca gacggaggtt tagagggaag aaacgatatg gctttgaagt tagtcggtgg
     4681 ttcacactta atagctaacg ccaagactac ttacaggtct aagaaaccag caaagaattt
     4741 gaaaatgcca ggagtttatt atgttgacta cagattggaa aggatcaagg aggccaataa
     4801 cgagacatat gtagagcagc atgaagtcgc cgtcgccaga tattgcgact taccttctaa
     4861 gttgggtcat aaattgaatg gatccggttc tccaccaaag aagaagagaa aggttggttc
     4921 ttaactcgag tggcagcttt tgattaagcc ttctagtcca aaaaacacgt ttttttgtca
     4981 tttatttcat tttcttagaa tagtttagtt tattcatttt atagtcacga atgttttatg
     5041 attctatata gggttgcaaa caagcatttt tcattttatg ttaaaacaat ttcaggttta
     5101 ccttttattc tgcttgtggt gacgcgtgta tccgcccgct cttttggtca cccatgtatg
     5161 ctgagcagtt acagagatgt tacgaaccac tagtgcactg cagtacacgg tttccttgaa
     5221 atttttttga ttcggtaatc tccgaacaga aggaagaacg aaggaaggag cacagactta
     5281 gattggtata tatacgcata tgtagtgttg aagaaacatg aaattgccca gtattcttaa
     5341 cccaactgca cagaacaaaa acgtgcagga aacgaagata aatcatgtcg aaagctacat
     5401 ataaggaacg tgctgctact catcctagtc ctgttgctgc caagctattt aatatcatgc
     5461 acgaaaagca aacaaacttg tgtgcttcat tggatgttcg taccaccaag gaattactgg
     5521 agttagttga agcattaggt cccaaaattt gtttactaaa aacacatgtg gatatcttga
     5581 ctgatttttc catggagggc acagttaagc cgctaaaggc attatccgcc aagtacaatt
     5641 ttttactctt cgaagacaga aaatttgctg acattggtaa tacagtcaaa ttgcagtact
     5701 ctgcgggtgt atacagaata gcagaatggg cagacattac gaatgcacac ggtgtggtgg
     5761 gcccaggtat tgttagcggt ttgaagcagg cggcagaaga agtaacaaag gaacctagag
     5821 gccttttgat gttagcagaa ttgtcatgca agggctccct atctactgga gaatatacta
     5881 agggtactgt tgacattgcg aagagcgaca aagattttgt tatcggcttt attgctcaaa
     5941 gagacatggg tggaagagat gaaggttacg attggttgat tatgacaccc ggtgtgggtt
     6001 tagatgacaa gggagatgca ttgggtcaac agtatagaac cgtggatgat gtggtttcta
     6061 caggatctga cattattatt gttggaagag gactatttgc aaagggaagg gatgctaagg
     6121 tagagggtga acgttacaga aaagcaggct gggaagcata tttgagaaga tgcggccagc
     6181 aaaactaaaa aactgtatta taagtaaatg catgtatact aaactcacaa attagagctt
     6241 caatttaatt atatcagtta ttacccgagt agagcacttg aatccactgc cccgggaatc
     6301 tcggtcgtaa tgatttctat aatgacgaaa aaaaaaaaat tggaaagaaa aagcttcatg
     6361 gcctttataa aaaggaacta tccaatacct cgccagaacc aagtaacagt attttacggg
     6421 gcacaaatca agaacaataa gacaggactg taaagatgga cgcattgaac tccaaagaac
     6481 aacaagagtt ccaaaaagta gtggaacaaa agcaaatgaa ggatttcatg cgtttgtact
     6541 ctaatctggt agaaagatgt ttcacagact gtgtcaatga cttcacaaca tcaaagctaa
     6601 ccaataagga acaaacatgc atcatgaagt gctcagaaaa gttcttgaag catagcgaac
     6661 gtgtagggca gcgtttccaa gaacaaaacg ctgccttggg acaaggcttg ggccgataag
     6721 gtgtactggc gtatatatat ctaattatgt atctctggtg tagcccattt ttagcatgta
     6781 aatataaaga ccgagcggcc gcgtgttaca accaattaac caattctgat tagaaaaact
     6841 catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt
     6901 gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa
     6961 gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc
     7021 cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg
     7081 agaatggcaa aagcttatgc atttctttcc agacttgttc aacaggccag ccattacgct
     7141 cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga
     7201 ggcgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc
     7261 gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata
     7321 cctggaatgc tgttttcccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac
     7381 ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca
     7441 tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg
     7501 catcgggctt cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag
     7561 cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc ctggagcaag
     7621 acgtttcccg ttgaatatgg ctcataacac cccttgtatt actgtttatg taagcagaca
     7681 gttttattgt tcatgatgat atatttttat cttgtgcaat gtaacatcag agattttgag
     7741 acacaacgtg gctttgttga ataaatcgaa cttttgctga gttgaaggat cagtcatgac
     7801 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     7861 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     7921 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     7981 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     8041 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     8101 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     8161 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     8221 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     8281 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     8341 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     8401 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     8461 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     8521 ctttcctgcg ttatcccctg attctgtgga taaccgtgcg gccgccaatg ctaaattcga
     8581 gtgaaacaca ggaagatcag aaaatcctca tttcatccat attaacaata atttcaaatg
     8641 tttatttgca ttatttgaaa ctaggcaaga caagcaacga aacgtttttg aaaattttga
     8701 gtattttcaa taaatttgta gaggactcag atattgaaaa aaagctacag caattaatac
     8761 ttgataagaa gagtattgag aagggcaacg gttcatcatc tcatggatct gcacatgaac
     8821 aaacaccaga gtcaaacgac gttgaaattg aggctactgc gccaattgat gacaatacag
     8881 acgatgataa caaaccgaag ttatctgatg tagaaaagga ttaaagatgc taagagatag
     8941 tgatgatatt tcataaataa tgtaattcta tatatgttaa ttaccttttt tgcgaggcat
     9001 atttatggtg aaggataagt tttgaccatc aaagaaggtt aatgtggctg tggtttcagg
     9061 gtccataaag cccacatgga taacattacc cctgaattcg catctagact ga
//
