LOCUS       sequence-250001-        9517 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of Venus-degronSwitchA and 
            mScarlet-degronSwitchC.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9517)
  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 9517)
  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: pAN1935
FEATURES             Location/Qualifiers
     source          1..9517
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        299..712
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             732..1445
                     /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             5790..6593
                     /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(7235..8050)
                     /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(7235..8050)
                     /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     7958..7977
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     8042..8061
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      8275..8863
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8764..8783
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 actggccgat aattgcagac gaacgccttg ccaacaggga gttcttcaga gacatggagg
       61 ctcaaaacga aattattgac agcctagaca tcaatagtca tacaacagaa agcgaccacc
      121 caactttggc tgataatagc gtataaacaa tgcatacttt gtacgttcaa aatacaatgc
      181 agtagatata tttatgcata ttacatataa tacatatcac ataggaagca acaggcgcgt
      241 tggactttta attttcgagg accgcgaatc cttacatcac acccaatccc ccacaagtga
      301 tcccccacac accatagctt caaaatgttt ctactccttt tttactcttc cagattttct
      361 cggactccgc gcatcgccgt accacttcaa aacacccaag cacagcatac taaatttccc
      421 ctctttcttc ctctagggtg tcgttaatta cccgtactaa aggtttggaa aagaaaaaag
      481 acaccgcctc gtttcttttt cttcgtcgaa aaaggcaata aaaattttta tcacgtttct
      541 ttttcttgaa aatttttttt tttgattttt ttctctttcg atgacctccc attgatattt
      601 aagttaataa acggtcatca atttctcaag tttcagtttc atttttcttg ttctattaca
      661 acttttttta cttcttgctc attagaaaga aagcatagca atctaatcta agttttaatt
      721 acaaaagatc tatgtctaaa ggtgaagaat tattcactgg tgttgtccca attttggttg
      781 aattagatgg tgatgttaat ggtcacaaat tttctgtctc cggtgaaggt gaaggtgatg
      841 ctacttacgg taaattgacc ttaaaattga tttgtactac tggtaaattg ccagttccat
      901 ggccaacctt agtcactact ttaggttatg gtttgcaatg ttttgctaga tacccagatc
      961 atatgaaaca acatgacttt ttcaagtctg ccatgccaga aggttatgtt caagaaagaa
     1021 ctattttttt caaagatgac ggtaactaca agaccagagc tgaagtcaag tttgaaggtg
     1081 ataccttagt taatagaatc gaattaaaag gtattgattt taaagaaggt ggtaacattt
     1141 taggtcacaa attggaatac aactataact ctcacaatgt ttacatcact gctgacaaac
     1201 aaaagaatgg tatcaaagct aacttcaaaa ttagacacaa cattgaagat ggtggtgttc
     1261 aattagctga ccattatcaa caaaatactc caattggtga tggtccagtc ttgttaccag
     1321 acaaccatta cttatcctat caatctgcct tatccaaaga tccaaacgaa aagagagatc
     1381 acatggtctt gttagaattt gttactgctg ctggtattac ccatggtatg gatgaattgt
     1441 acaaaggatc cagtaaggaa gcagtgacga agctacaggc ccttaacatc aaattggcgg
     1501 aaaaattgct tgaagctgtc acgaagttgc aagcattaaa cataaaactt gccgaaaagc
     1561 tgttagaagc cctggcacgt ctgcaagagc tgaacattgc cttggtttat ttagccgtag
     1621 aattaacgga ccctaagaga attgcggacg agattaaaaa agttaaagac aagagcaagg
     1681 aaattgtcga gagagcggaa gaggagatag ctagggcggc agccgagtcc aagaaaattt
     1741 tagacgaagc cgaggaggaa atcgcccgtg cagcagcaga gtccaagaag atccttgatg
     1801 aaggatccgg aagtggttca gacgcggtag cggaattaca ggcattaaat ctgaagctag
     1861 ccgagttact tctggaggct gtggctgagc ttcaagcgct taacctgaaa ctagcggagc
     1921 tacttttgga ggctatagct aagctacaag agctgaatat taagcttgtc gagctattga
     1981 ctaaactaac tgaccctgct accatcaggg aggcgatcag aaaggtgaag gaagactccg
     2041 agaggatagt ggcggaggca gagcgtttga tagccgctgc gaaggctgaa agtgaacgta
     2101 ttatacgtga agctgagaga ttgatagctg ctgctaaagc ggagtccgag aggataatta
     2161 gagaggggtc cggttcaggg gaccctgacg tagcgagact gcaagaacta aatatcgagc
     2221 tagctagaga gctgctaagg gatgtcgcta gactacaaga gctaaacata gaactggcga
     2281 gagaacttct aagggcagcg gcagagttac aagagctgaa cataaagtta gtcgagctag
     2341 cgagtgagtt aaccgatcct gatgaagcac gtaaagcaat cgccagggta aagagagaga
     2401 gtaaaagaat cgttgaggat cttattatga gctgtgccca agagtctgct gcaagcgaga
     2461 agattagcag agaagccgaa agactgataa ggtaatggca gtgcttttaa ctaagaatta
     2521 ttagtctttt ctgcttattt tttcatcata gtttagaaca ctttatatta acgaatagtt
     2581 tatgaatcta tttaggttta aaaattgata cagttttata agttactttt tcaaagactc
     2641 gtgctgtcta ttgcataatg cactggaagg ggaaaaaaaa ggtgcacacg cgtggctttt
     2701 tcttgaattt gcagtttgaa aaatgctgcc aaaccagatg tcaacacagc tacaacgttg
     2761 ataggtcaag atcaatgtaa acaattactt tgttatgtag agttttttta gctacctata
     2821 ttccaccata acatcaatca tgcggttgct ggtgtattta ccaataatgt ttaatgtata
     2881 tatatatata tatatatggg gccgtatact tacatatagt agatgtcaag cgtaggcgct
     2941 tcccctgccg gctgtgaggg cgccataacc aaggtatcta tagaccgcca atcagcaaac
     3001 tacctccgta cattcatgtt gcacccacac atttatacac ccagaccgcg acaaattacc
     3061 cataaggttg tttgtgacgg cgtcgtacaa gagaacgtgg gaacttttta ggctcaccaa
     3121 aaaagaaaga aaaaatacga gttgctgaca gaagcctcaa gaaaaaaaaa attcttcttc
     3181 gactatgctg gaggcagaga tgatcgagcc ggtagttaac tatatatagc taaattggtt
     3241 ccatcacctt cttttctggt gtcgctcctt ctagtgctat ttctggcttt tcctattttt
     3301 ttttttccat ttttctttct ctctttctaa tatataaatt ctcttgcatt ttctattttt
     3361 ctctctatct attctacttg tttattccct tcaaggtttt tttttaagga gtacttgttt
     3421 ttagaatata cggtcaacga actataatta actaaacaga tctatggtat caaaaggcga
     3481 ggcggtcatt aaagagttca tgaggtttaa agtccatatg gaaggaagca tgaatggtca
     3541 cgagtttgag atagagggag agggtgaggg aaggccgtac gaagggacgc aaactgcgaa
     3601 gttgaaggta acgaaaggcg gccctctgcc attctcttgg gacatccttt caccgcaatt
     3661 tatgtacggc tctagggcct tcacaaaaca ccccgccgat attccggatt actataaaca
     3721 aagttttcct gagggcttca agtgggaaag agtaatgaac ttcgaggatg ggggagctgt
     3781 gacggttact caagacacta gcctggagga cggtactctt atatacaagg taaaactaag
     3841 agggacgaat tttccgccag acggccccgt gatgcaaaaa aaaactatgg ggtgggaagc
     3901 atccaccgag aggctgtacc ccgaagacgg ggtcctgaag ggtgatatta aaatggcatt
     3961 aagattaaaa gatggcggga gatacttagc tgacttcaag actacatata aagccaaaaa
     4021 acctgtacag atgcctggag cttacaatgt ggatcgtaag ctggatatta catcccacaa
     4081 cgaagattat accgtggtgg agcagtacga gcgttcagag gggcgtcact ccaccggggg
     4141 tatggatgaa ttatacaaga gcggtcttag aagcagggcg caagcgtcaa attcagcggt
     4201 agatggaaca gcaggtccag gtagtactgg cagcagagga tctgggtcgg gttcaggctc
     4261 tggatcggga tcctctctgg aggcggttct taaactggcg gagttaaatc taaaacttag
     4321 cgataagcta gccgaagctg tccaaaaact agcagcccta ttgaacaaac tgcttgagaa
     4381 gttgtccgag gccctgcaga ggcttttcga attgaacgtt gccttggtaa cccttgcgat
     4441 tgaattgacc gaccctaaga ggatagcgga tgagatcaaa aaggtgaaag ataagagtaa
     4501 ggagatcgtg gaaagggccg aggaggagat tgccagagcg gcggcagaga gtaagaaaat
     4561 tttggacgag gccgaggaag aaattgcgag agcagcggcg gaatcaaaga aaatcttgga
     4621 cgaaggcagt ggcagtggat ctgacgcagt tgcagaactt caggccctga atttaaaatt
     4681 agcagaattg ctgctggagg cagtagcaga gctacaggcg ttaaatctaa aactggcgga
     4741 attgctactg gaggcgatag ctaaactaca agaattgaat atcaagctgg tagagctgtt
     4801 gactaaatta acggaccctg caacaataag agaggctata agaaaagtga aagaagattc
     4861 cgaacgtatt gttgcagaag ccgagcgttt gatcgctgct gcgaaagcgg aatctgaaag
     4921 gattatccgt gaagcagaac gtttgatcgc cgccgcaaag gcagagtcag aaagaataat
     4981 tcgtgaagga tcaggaagta atgatccgtt agtagcaaga cttcaagaat tgcttataga
     5041 acacgcacgt gaacttctaa gactggtcgc gacatctcag gagatattca tagaattggc
     5101 cagagcattt ctagctaatg ctgctcagtt acaagaggcc gcaataaaag ccgtcgaggc
     5161 ggcatcagaa aacgggtccg ggtccggtcc gagctccgag aaggtgagga gagaattgaa
     5221 agaatctttg aaagagaacc acaaacaaaa tcagaaactt ttaatgagct gtgcacagga
     5281 gcaagagaag ctgaataggg aactagagga attgaaaaaa aaacataaaa aataatggca
     5341 ttgaattgaa ttgaaatcga tagatcaatt tttttctttt ctctttcccc atcctttacg
     5401 ctaaaataat agtttatttt attttttgaa tattttttat ttatatacgt atatatagac
     5461 tattatttat cttttaatga ttattaagat ttttattaaa aaaaaattcg ctcctctttt
     5521 aatgccttta tgcagttttt ttttcccatt cgatatttct atgtgctgag cagttacaga
     5581 gatgttacga accactagtg cactgcagta cacggtttcc ttgaaatttt tttgattcgg
     5641 taatctccga acagaaggaa gaacgaagga aggagcacag acttagattg gtatatatac
     5701 gcatatgtag tgttgaagaa acatgaaatt gcccagtatt cttaacccaa ctgcacagaa
     5761 caaaaacgtg caggaaacga agataaatca tgtcgaaagc tacatataag gaacgtgctg
     5821 ctactcatcc tagtcctgtt gctgccaagc tatttaatat catgcacgaa aagcaaacaa
     5881 acttgtgtgc ttcattggat gttcgtacca ccaaggaatt actggagtta gttgaagcat
     5941 taggtcccaa aatttgttta ctaaaaacac atgtggatat cttgactgat ttttccatgg
     6001 agggcacagt taagccgcta aaggcattat ccgccaagta caatttttta ctcttcgaag
     6061 acagaaaatt tgctgacatt ggtaatacag tcaaattgca gtactctgcg ggtgtataca
     6121 gaatagcaga atgggcagac attacgaatg cacacggtgt ggtgggccca ggtattgtta
     6181 gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc tagaggcctt ttgatgttag
     6241 cagaattgtc atgcaagggc tccctatcta ctggagaata tactaagggt actgttgaca
     6301 ttgcgaagag cgacaaagat tttgttatcg gctttattgc tcaaagagac atgggtggaa
     6361 gagatgaagg ttacgattgg ttgattatga cacccggtgt gggtttagat gacaagggag
     6421 atgcattggg tcaacagtat agaaccgtgg atgatgtggt ttctacagga tctgacatta
     6481 ttattgttgg aagaggacta tttgcaaagg gaagggatgc taaggtagag ggtgaacgtt
     6541 acagaaaagc aggctgggaa gcatatttga gaagatgcgg ccagcaaaac taaaaaactg
     6601 tattataagt aaatgcatgt atactaaact cacaaattag agcttcaatt taattatatc
     6661 agttattacc cgagtagagc acttgaatcc actgccccgg gaatctcggt cgtaatgatt
     6721 tctataatga cgaaaaaaaa aaaattggaa agaaaaagct tcatggcctt tataaaaagg
     6781 aactatccaa tacctcgcca gaaccaagta acagtatttt acggggcaca aatcaagaac
     6841 aataagacag gactgtaaag atggacgcat tgaactccaa agaacaacaa gagttccaaa
     6901 aagtagtgga acaaaagcaa atgaaggatt tcatgcgttt gtactctaat ctggtagaaa
     6961 gatgtttcac agactgtgtc aatgacttca caacatcaaa gctaaccaat aaggaacaaa
     7021 catgcatcat gaagtgctca gaaaagttct tgaagcatag cgaacgtgta gggcagcgtt
     7081 tccaagaaca aaacgctgcc ttgggacaag gcttgggccg ataaggtgta ctggcgtata
     7141 tatatctaat tatgtatctc tggtgtagcc catttttagc atgtaaatat aaagaccgag
     7201 cggccgcgtg ttacaaccaa ttaaccaatt ctgattagaa aaactcatcg agcatcaaat
     7261 gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa agccgtttct
     7321 gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt
     7381 ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa
     7441 ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagct
     7501 tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac
     7561 tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgaggcga aatacgcgat
     7621 cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca
     7681 gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt
     7741 tcccggggat cgcagtggtg agtaaccatg catcatcagg agtacggata aaatgcttga
     7801 tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacat
     7861 cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg ggcttcccat
     7921 acaatcgata gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat
     7981 ataaatcagc atccatgttg gaatttaatc gcggcctgga gcaagacgtt tcccgttgaa
     8041 tatggctcat aacacccctt gtattactgt ttatgtaagc agacagtttt attgttcatg
     8101 atgatatatt tttatcttgt gcaatgtaac atcagagatt ttgagacaca acgtggcttt
     8161 gttgaataaa tcgaactttt gctgagttga aggatcagtc atgaccaaaa tcccttaacg
     8221 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     8281 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     8341 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     8401 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     8461 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     8521 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     8581 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     8641 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     8701 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     8761 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     8821 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     8881 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     8941 ccctgattct gtggataacc gtgcggccgc caatgctaaa ttcgagtgaa acacaggaag
     9001 atcagaaaat cctcatttca tccatattaa caataatttc aaatgtttat ttgcattatt
     9061 tgaaactagg caagacaagc aacgaaacgt ttttgaaaat tttgagtatt ttcaataaat
     9121 ttgtagagga ctcagatatt gaaaaaaagc tacagcaatt aatacttgat aagaagagta
     9181 ttgagaaggg caacggttca tcatctcatg gatctgcaca tgaacaaaca ccagagtcaa
     9241 acgacgttga aattgaggct actgcgccaa ttgatgacaa tacagacgat gataacaaac
     9301 cgaagttatc tgatgtagaa aaggattaaa gatgctaaga gatagtgatg atatttcata
     9361 aataatgtaa ttctatatat gttaattacc ttttttgcga ggcatattta tggtgaagga
     9421 taagttttga ccatcaaaga aggttaatgt ggctgtggtt tcagggtcca taaagcccac
     9481 atggataaca ttacccctga attcgcatct agactga
//
