LOCUS       sequence-249999-        9021 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dual inducible expression of SynTF-mScarlet-nesSwitchA and 
            KeyA-BFP-NLS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9021)
  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 9021)
  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: pAN1777
FEATURES             Location/Qualifiers
     source          1..9021
                     /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             5294..6097
                     /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(6739..7554)
                     /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(6739..7554)
                     /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     7462..7481
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     7546..7565
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      7779..8367
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8268..8287
                     /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 atctatggcc gctgcgaaga gggcgagaac tagtggaggg
      601 tctggttctg gggtcgacgg tggactgcag gccccaccaa ccgatgtttc tttaggtgat
      661 gagttacatt tggacggtga ggacgtagct atggcacatg cagatgcctt agacgatttt
      721 gacttggata tgttaggaga tggagattca ccaggacctg gtttcacacc acacgattca
      781 gctccatacg gtgctttgga catggccgat ttcgaatttg aacagatgtt tactgatgcc
      841 ttgggtatag acgaatacgg tggtggagct agacccgggg agcgcccctt ccagtgtcgc
      901 atttgcatgc ggaacttttc gcgccaggac aggcttgaca ggcatacccg tactcatacc
      961 ggtgaaaaac cgtttcagtg tcggatctgt atgcgaaatt tctcccagaa ggagcacttg
     1021 gcggggcatc tacgtacgca caccggcgag aagccattcc aatgccgaat atgcatgcgc
     1081 aacttcagtc gccgcgacaa cctgaaccgg cacctaaaaa cccacctgag aggatctgga
     1141 tccggttctg tatcaaaagg cgaggcggtc attaaagagt tcatgaggtt taaagtccat
     1201 atggaaggaa gcatgaatgg tcacgagttt gagatagagg gagagggtga gggaaggccg
     1261 tacgaaggga cgcaaactgc gaagttgaag gtaacgaaag gcggccctct gccattctct
     1321 tgggacatcc tttcaccgca atttatgtac ggctctaggg ccttcacaaa acaccccgcc
     1381 gatattccgg attactataa acaaagtttt cctgagggct tcaagtggga aagagtaatg
     1441 aacttcgagg atgggggagc tgtgacggtt actcaagaca ctagcctgga ggacggtact
     1501 cttatataca aggtaaaact aagagggacg aattttccgc cagacggccc cgtgatgcaa
     1561 aaaaaaacta tggggtggga agcatccacc gagaggctgt accccgaaga cggggtcctg
     1621 aagggtgata ttaaaatggc attaagatta aaagatggcg ggagatactt agctgacttc
     1681 aagactacat ataaagccaa aaaacctgta cagatgcctg gagcttacaa tgtggatcgt
     1741 aagctggata ttacatccca caacgaagat tataccgtgg tggagcagta cgagcgttca
     1801 gaggggcgtc actccaccgg gggtatggat gaattataca agagcggtct tagaagcagg
     1861 gcgcaagcgt caaattcagc ggtagatgga acagcaggtc caggtagtac tggcagcaga
     1921 ggatccagta aggaagcagt gacgaagcta caggccctta acatcaaatt ggcggaaaaa
     1981 ttgcttgaag ctgtcacgaa gttgcaagca ttaaacataa aacttgccga aaagctgtta
     2041 gaagccctgg cacgtctgca agagctgaac attgccttgg tttatttagc cgtagaatta
     2101 acggacccta agagaattgc ggacgagatt aaaaaagtta aagacaagag caaggaaatt
     2161 gtcgagagag cggaagagga gatagctagg gcggcagccg agtccaagaa aattttagac
     2221 gaagccgagg aggaaatcgc ccgtgcagca gcagagtcca agaagatcct tgatgaagga
     2281 tccggaagtg gttcagacgc ggtagcggaa ttacaggcat taaatctgaa gctagccgag
     2341 ttacttctgg aggctgtggc tgagcttcaa gcgcttaacc tgaaactagc ggagctactt
     2401 ttggaggcta tagctaagct acaagagctg aatattaagc ttgtcgagct attgactaaa
     2461 ctaactgacc ctgctaccat cagggaggcg atcagaaagg tgaaggaaga ctccgagagg
     2521 atagtggcgg aggcagagcg tttgatagcc gctgcgaagg ctgaaagtga acgtattata
     2581 cgtgaagctg agagattgat agctgctgct aaagcggagt ccgagaggat aattagagag
     2641 gggtccggtt caggggaccc tgacgtagcg agactgcaag aactaaatat cgagctagct
     2701 agagagctgc taagggatgt cgctagacta caagagctaa acatagaact ggcgagagaa
     2761 cttctaaggg cagcggcaga gttacaagag ctgaacataa agttagtcga gctagcgagt
     2821 gagttaaccg atcctgatga agcacgtaaa gcaatcgcca gggtaaagag agagagtaaa
     2881 agaatagtcg aagacttggc gctaaaatta gcaggattgg atataaatag cgaaaaaatc
     2941 agcagagaag cagagagact aatccgtgag gccgcataat ggcgccaatt ggtgcggcaa
     3001 ttgataataa cgaaaatgtc ttttaatgat ctgggtataa tgaggaattt tccgaacgtt
     3061 tttactttat atatatatat acatgtaaca tatattctat acgctataga gaaaggaaat
     3121 ttttcaatta aaaaaaaaat agagaaagag tttcacttct tgattatcgc taacactaat
     3181 ggttgaagta ctgctacttt aattttatgc tgccaaacca gatgtcaaca cagctacaac
     3241 gttatattga attttcaaaa attcttactt tttttttgga tggacgcaaa gaagtttaat
     3301 aatcatatta catggcatta ccaccatata catatccata tacatatcca tatctaatct
     3361 tacttatatg ttgtggaaat gtaaagagcc ccattatctt agcctaaaaa aaccttctct
     3421 ttggaacttt cagtaatacg cttaactgct cattgctata ttgaagtgtg gccgcgtggg
     3481 cgtgcgtggg cgggcgtggg cgtgcgtggg cgggcgtggg cgtgcgtggg cgtctagacc
     3541 gtgcgtcctc gtcttcaccg gtcgcgttcc tgaaacgcag atgtgcctaa tgccgcactg
     3601 ctccgaacaa taaagattct acaatactag cttttatggt tatgaagagg aaaaattggc
     3661 agtaacctgg ccccacaaac cttcaaatta acgaatcaaa ttaacaacca taggatgata
     3721 atgcgattag ttttttagcc ttatttctgg ggtaattaat cagcgaagcg atgatttttg
     3781 atctattaac agatatataa atggaaaagc tgcataacca ctttaactaa tactttcaac
     3841 attttcagtt tgtattactt cttattcaaa tgtcataaaa gtatcaacaa aaaattgtta
     3901 atatacctct atactttaac gtcaaggaga aaaaactata agatctatgg aggctagaaa
     3961 agccatcgct agagtaaaga gagaatcaaa gagaatcgta gaagatgctg agaggttaat
     4021 aagagaagca gccgccgcat ctgagaaaat ttcaagggag gccgaaaggt tgattaggga
     4081 ggcagctgcg gcttcagaaa agataagtag ggagggttct gtttcaaagg gtgaagaatt
     4141 gatcaaagaa aacatgcata tgaaattata tatggaaggt acagttgaca accatcattt
     4201 taagtgtaca tctgaaggag aaggtaagcc ttacgaggga actcaaacta tgagaataaa
     4261 agtcgtagaa ggtggaccat tgccttttgc ttttgatatc ttagcaacat ctttcttata
     4321 tggttctaaa acctttatta accatactca aggtatccct gatttcttca aacaatcttt
     4381 ccctgaagga tttacttggg agagggttac aacatatgag gatggtggtg ttttgacagc
     4441 aacccaggat acatcattgc aggatggatg tttgatttat aatgtaaaga ttagaggagt
     4501 aaattttact tctaacggtc ctgtaatgca aaagaagact ttaggttggg aagctttcac
     4561 cgaaacatta tatccagcag acggaggttt agagggaaga aacgatatgg ctttgaagtt
     4621 agtcggtggt tcacacttaa tagctaacgc caagactact tacaggtcta agaaaccagc
     4681 aaagaatttg aaaatgccag gagtttatta tgttgactac agattggaaa ggatcaagga
     4741 ggccaataac gagacatatg tagagcagca tgaagtcgcc gtcgccagat attgcgactt
     4801 accttctaag ttgggtcata aattgaatgg atcctaactc gagagctttt gattaagcct
     4861 tctagtccaa aaaacacgtt tttttgtcat ttatttcatt ttcttagaat agtttagttt
     4921 attcatttta tagtcacgaa tgttttatga ttctatatag ggttgcaaac aagcattttt
     4981 cattttatgt taaaacaatt tcaggtttac cttttattct gcttgtggtg acgcgtgtat
     5041 ccgcccgctc ttttggtcac ccatgtatgc tgagcagtta cagagatgtt acgaaccact
     5101 agtgcactgc agtacacggt ttccttgaaa tttttttgat tcggtaatct ccgaacagaa
     5161 ggaagaacga aggaaggagc acagacttag attggtatat atacgcatat gtagtgttga
     5221 agaaacatga aattgcccag tattcttaac ccaactgcac agaacaaaaa cgtgcaggaa
     5281 acgaagataa atcatgtcga aagctacata taaggaacgt gctgctactc atcctagtcc
     5341 tgttgctgcc aagctattta atatcatgca cgaaaagcaa acaaacttgt gtgcttcatt
     5401 ggatgttcgt accaccaagg aattactgga gttagttgaa gcattaggtc ccaaaatttg
     5461 tttactaaaa acacatgtgg atatcttgac tgatttttcc atggagggca cagttaagcc
     5521 gctaaaggca ttatccgcca agtacaattt tttactcttc gaagacagaa aatttgctga
     5581 cattggtaat acagtcaaat tgcagtactc tgcgggtgta tacagaatag cagaatgggc
     5641 agacattacg aatgcacacg gtgtggtggg cccaggtatt gttagcggtt tgaagcaggc
     5701 ggcagaagaa gtaacaaagg aacctagagg ccttttgatg ttagcagaat tgtcatgcaa
     5761 gggctcccta tctactggag aatatactaa gggtactgtt gacattgcga agagcgacaa
     5821 agattttgtt atcggcttta ttgctcaaag agacatgggt ggaagagatg aaggttacga
     5881 ttggttgatt atgacacccg gtgtgggttt agatgacaag ggagatgcat tgggtcaaca
     5941 gtatagaacc gtggatgatg tggtttctac aggatctgac attattattg ttggaagagg
     6001 actatttgca aagggaaggg atgctaaggt agagggtgaa cgttacagaa aagcaggctg
     6061 ggaagcatat ttgagaagat gcggccagca aaactaaaaa actgtattat aagtaaatgc
     6121 atgtatacta aactcacaaa ttagagcttc aatttaatta tatcagttat tacccgagta
     6181 gagcacttga atccactgcc ccgggaatct cggtcgtaat gatttctata atgacgaaaa
     6241 aaaaaaaatt ggaaagaaaa agcttcatgg cctttataaa aaggaactat ccaatacctc
     6301 gccagaacca agtaacagta ttttacgggg cacaaatcaa gaacaataag acaggactgt
     6361 aaagatggac gcattgaact ccaaagaaca acaagagttc caaaaagtag tggaacaaaa
     6421 gcaaatgaag gatttcatgc gtttgtactc taatctggta gaaagatgtt tcacagactg
     6481 tgtcaatgac ttcacaacat caaagctaac caataaggaa caaacatgca tcatgaagtg
     6541 ctcagaaaag ttcttgaagc atagcgaacg tgtagggcag cgtttccaag aacaaaacgc
     6601 tgccttggga caaggcttgg gccgataagg tgtactggcg tatatatatc taattatgta
     6661 tctctggtgt agcccatttt tagcatgtaa atataaagac cgagcggccg cgtgttacaa
     6721 ccaattaacc aattctgatt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
     6781 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
     6841 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
     6901 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
     6961 atcaccatga gtgacgactg aatccggtga gaatggcaaa agcttatgca tttctttcca
     7021 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
     7081 gttattcatt cgtgattgcg cctgagcgag gcgaaatacg cgatcgctgt taaaaggaca
     7141 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt
     7201 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt
     7261 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat
     7321 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
     7381 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt
     7441 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
     7501 gttggaattt aatcgcggcc tggagcaaga cgtttcccgt tgaatatggc tcataacacc
     7561 ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgatgata tatttttatc
     7621 ttgtgcaatg taacatcaga gattttgaga cacaacgtgg ctttgttgaa taaatcgaac
     7681 ttttgctgag ttgaaggatc agtcatgacc aaaatccctt aacgtgagtt ttcgttccac
     7741 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     7801 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     7861 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     7921 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     7981 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     8041 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     8101 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     8161 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     8221 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     8281 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     8341 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     8401 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     8461 aaccgtgcgg ccgccaatgc taaattcgag tgaaacacag gaagatcaga aaatcctcat
     8521 ttcatccata ttaacaataa tttcaaatgt ttatttgcat tatttgaaac taggcaagac
     8581 aagcaacgaa acgtttttga aaattttgag tattttcaat aaatttgtag aggactcaga
     8641 tattgaaaaa aagctacagc aattaatact tgataagaag agtattgaga agggcaacgg
     8701 ttcatcatct catggatctg cacatgaaca aacaccagag tcaaacgacg ttgaaattga
     8761 ggctactgcg ccaattgatg acaatacaga cgatgataac aaaccgaagt tatctgatgt
     8821 agaaaaggat taaagatgct aagagatagt gatgatattt cataaataat gtaattctat
     8881 atatgttaat tacctttttt gcgaggcata tttatggtga aggataagtt ttgaccatca
     8941 aagaaggtta atgtggctgt ggtttcaggg tccataaagc ccacatggat aacattaccc
     9001 ctgaattcgc atctagactg a
//
