LOCUS       sequence-431863-        6283 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express degron assay: Degradation tag-YFP-ePTS1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6283)
  AUTHORS   Baker JJ, Shi J, Wang S, Mujica EM, Bianco S, Capponi S, Dueber JE
  TITLE     ML-enhanced peroxisome capacity enables compartmentalization of 
            multienzyme pathway.
  JOURNAL   Nat Chem Biol. 2024 Oct 14. doi: 10.1038/s41589-024-01759-2.
  PUBMED    39402374
REFERENCE   2  (bases 1 to 6283)
  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: "Nat Chem 
            Biol. 2024 Oct 14. doi: 10.1038/s41589-024-01759-2."
COMMENT     Sequence Label: pJJB207
FEATURES             Location/Qualifiers
     source          1..6283
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        14..680
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     550..573
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     CDS             687..914
                     /codon_start=1
                     /product="Saccharomyces cerevisiae ubiquitin"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVESSDTIDNVKSKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     CDS             1041..1751
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKL
                     ICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEGGNILGHKLEYNYNSHNVYITADKQKNGIKAN
                     FKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     CDS             2402..3205
                     /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(3847..4662)
                     /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(3847..4662)
                     /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     4570..4589
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4654..4673
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4887..5475
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5376..5395
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cagttcgagt ttatcattat caatactgcc atttcaaaga atacgtaaat aattaatagt
       61 agtgattttc ctaactttat ttagtcaaaa aattagcctt ttaattctgc tgtaacccgt
      121 acatgcccaa aatagggggc gggttacaca gaatatataa catcgtaggt gtctgggtga
      181 acagtttatt cctggcatcc actaaatata atggagcccg ctttttaagc tggcatccag
      241 aaaaaaaaag aatcccagca ccaaaatatt gttttcttca ccaaccatca gttcataggt
      301 ccattctctt agcgcaacta cagagaacag gggcacaaac aggcaaaaaa cgggcacaac
      361 ctcaatggag tgatgcaacc tgcctggagt aaatgatgac acaaggcaat tgacccacgc
      421 atgtatctat ctcattttct tacaccttct attaccttct gctctctctg atttggaaaa
      481 agctgaaaaa aaaggttgaa accagttccc tgaaattatt cccctacttg actaataagt
      541 atataaagac ggtaggtatt gattgtaatt ctgtaaatct atttcttaaa cttcttaaat
      601 tctactttta tagttagtct tttttttagt tttaaaacac caagaactta gtttcgaata
      661 aacacacata aacaaacaaa agatctatgc agattttcgt taaaacttta actggtaaaa
      721 caattacttt agaagttgag tcatctgata ctatcgataa tgttaaatct aagatccagg
      781 ataaggaagg tattcctcca gatcaacaaa gattgatttt cgcaggtaag cagttggagg
      841 acggtaggac tttgtctgat tataacatac agaaagaatc tactttgcac ttagtcttaa
      901 gattaagggg aggttatcat ggttctggtg cctggttgtt acctgtatca ttggttaaaa
      961 gaaagaccac attggcacca aatactcaaa ctgcctctcc aagagccttg gcagactctt
     1021 taatgcagag atcaggttct tctaaaggtg aagaattatt cactggtgtt gtcccaattt
     1081 tggttgaatt agatggtgat gttaatggtc acaaattttc tgtctccggt gaaggtgaag
     1141 gtgatgctac ttacggtaaa ttgaccttaa aattgatttg tactactggt aaattgccag
     1201 ttccatggcc aaccttagtc actactttag gttatggttt gcaatgtttt gctagatacc
     1261 cagatcatat gaaacaacat gactttttca agtctgccat gccagaaggt tatgttcaag
     1321 aaagaactat ttttttcaaa gatgacggta actacaagac cagagctgaa gtcaagtttg
     1381 aaggtgatac cttagttaat agaatcgaat taaaaggtat tgattttaaa gaaggtggta
     1441 acattttagg tcacaaattg gaatacaact ataactctca caatgtttac atcactgctg
     1501 acaaacaaaa gaatggtatc aaagctaact tcaaaattag acacaacatt gaagatggtg
     1561 gtgttcaatt agctgaccat tatcaacaaa atactccaat tggtgatggt ccagtcttgt
     1621 taccagacaa ccattactta tcctatcaat ctgccttatc caaagatcca aacgaaaaga
     1681 gagatcacat ggtcttgtta gaatttgtta ctgctgctgg tattacccat ggtatggatg
     1741 aattgtacaa agggtccttg ggaagaggta gaagatccaa attgtaagga tcctaactcg
     1801 agataaagca atcttgatga ggataatgat ttttttttga atatacataa atactaccgt
     1861 ttttctgcta gattttgtga tgacgtaaat aagtacatat tactttttaa gccaagacaa
     1921 gattaagcat taactttacc cttttctttc taagtttcaa tattagttat cactgtttaa
     1981 aagttatggc gagaacgtcg gcggttaaaa tatattaccc tgaacggctg gaaatctgct
     2041 cgtcagtggt gctcacactg acgaatcatg tacagatcat accgatgact gcctggcgac
     2101 tcacaactaa gcaagacagc cggaaccagc gccggcgaac accactgcat atatggcata
     2161 tcacaacagt ccacgtctca agcagttaca gagatgttac gaaccactag tgcactgcag
     2221 tacacggttt ccttgaaatt tttttgattc ggtaatctcc gaacagaagg aagaacgaag
     2281 gaaggagcac agacttagat tggtatatat acgcatatgt agtgttgaag aaacatgaaa
     2341 ttgcccagta ttcttaaccc aactgcacag aacaaaaacg tgcaggaaac gaagataaat
     2401 catgtcgaaa gctacatata aggaacgtgc tgctactcat cctagtcctg ttgctgccaa
     2461 gctatttaat atcatgcacg aaaagcaaac aaacttgtgt gcttcattgg atgttcgtac
     2521 caccaaggaa ttactggagt tagttgaagc attaggtccc aaaatttgtt tactaaaaac
     2581 acatgtggat atcttgactg atttttccat ggagggcaca gttaagccgc taaaggcatt
     2641 atccgccaag tacaattttt tactcttcga agacagaaaa tttgctgaca ttggtaatac
     2701 agtcaaattg cagtactctg cgggtgtata cagaatagca gaatgggcag acattacgaa
     2761 tgcacacggt gtggtgggcc caggtattgt tagcggtttg aagcaggcgg cagaagaagt
     2821 aacaaaggaa cctagaggcc ttttgatgtt agcagaattg tcatgcaagg gctccctatc
     2881 tactggagaa tatactaagg gtactgttga cattgcgaag agcgacaaag attttgttat
     2941 cggctttatt gctcaaagag acatgggtgg aagagatgaa ggttacgatt ggttgattat
     3001 gacacccggt gtgggtttag atgacaaggg agatgcattg ggtcaacagt atagaaccgt
     3061 ggatgatgtg gtttctacag gatctgacat tattattgtt ggaagaggac tatttgcaaa
     3121 gggaagggat gctaaggtag agggtgaacg ttacagaaaa gcaggctggg aagcatattt
     3181 gagaagatgc ggccagcaaa actaaaaaac tgtattataa gtaaatgcat gtatactaaa
     3241 ctcacaaatt agagcttcaa tttaattata tcagttatta cccgagtaga gcacttgaat
     3301 ccactgcccc gggaatctcg gtcgtaatga tttctataat gacgaaaaaa aaaaaattgg
     3361 aaagaaaaag cttcatggcc tttataaaaa ggaactatcc aatacctcgc cagaaccaag
     3421 taacagtatt ttacggggca caaatcaaga acaataagac aggactgtaa agatggacgc
     3481 attgaactcc aaagaacaac aagagttcca aaaagtagtg gaacaaaagc aaatgaagga
     3541 tttcatgcgt ttgtactcta atctggtaga aagatgtttc acagactgtg tcaatgactt
     3601 cacaacatca aagctaacca ataaggaaca aacatgcatc atgaagtgct cagaaaagtt
     3661 cttgaagcat agcgaacgtg tagggcagcg tttccaagaa caaaacgctg ccttgggaca
     3721 aggcttgggc cgataaggtg tactggcgta tatatatcta attatgtatc tctggtgtag
     3781 cccattttta gcatgtaaat ataaagaccg agcggccgcg tgttacaacc aattaaccaa
     3841 ttctgattag aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt
     3901 atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca
     3961 gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat
     4021 acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt
     4081 gacgactgaa tccggtgaga atggcaaaag cttatgcatt tctttccaga cttgttcaac
     4141 aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg
     4201 tgattgcgcc tgagcgaggc gaaatacgcg atcgctgtta aaaggacaat tacaaacagg
     4261 aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc
     4321 aggatattct tctaatacct ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca
     4381 tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag
     4441 ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt
     4501 cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg cacctgattg
     4561 cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa
     4621 tcgcggcctg gagcaagacg tttcccgttg aatatggctc ataacacccc ttgtattact
     4681 gtttatgtaa gcagacagtt ttattgttca tgatgatata tttttatctt gtgcaatgta
     4741 acatcagaga ttttgagaca caacgtggct ttgttgaata aatcgaactt ttgctgagtt
     4801 gaaggatcag tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     4861 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     4921 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     4981 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     5041 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     5101 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     5161 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     5221 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     5281 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     5341 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     5401 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     5461 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     5521 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtgcggcc
     5581 gccaatgcta aattcgagtg aaacacagga agatcagaaa atcctcattt catccatatt
     5641 aacaataatt tcaaatgttt atttgcatta tttgaaacta ggcaagacaa gcaacgaaac
     5701 gtttttgaaa attttgagta ttttcaataa atttgtagag gactcagata ttgaaaaaaa
     5761 gctacagcaa ttaatacttg ataagaagag tattgagaag ggcaacggtt catcatctca
     5821 tggatctgca catgaacaaa caccagagtc aaacgacgtt gaaattgagg ctactgcgcc
     5881 aattgatgac aatacagacg atgataacaa accgaagtta tctgatgtag aaaaggatta
     5941 aagatgctaa gagatagtga tgatatttca taaataatgt aattctatat atgttaatta
     6001 ccttttttgc gaggcatatt tatggtgaag gataagtttt gaccatcaaa gaaggttaat
     6061 gtggctgtgg tttcagggtc cataaagccc acatggataa cattacccct gaattcgcat
     6121 ctagactgat gagacgtggt agagccacaa acagccggta caagcaacga tctccaggac
     6181 catctgaatc atgcgcggat gacacgaact cacgacggcg atcacagaca ttaacccaca
     6241 gtacagacac tgcgacaacg tggcaattcg tcgcaataca acg
//
