LOCUS       sequence-431865-        5179 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 5179)
  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 5179)
  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: pSKT001
FEATURES             Location/Qualifiers
     source          1..5179
                     /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"
     terminator      1801..1988
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1856..1880)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(2763..3578)
                     /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(2763..3578)
                     /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     3486..3505
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3570..3589
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3803..4391
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4292..4311
                     /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 aggatccggg tccttgggaa gaggtagaag atccaaattg tagtaatggc
     1801 gcgaatttct tatgatttat gatttttatt attaaataag ttataaaaaa aataagtgta
     1861 tacaaatttt aaagtgactc ttaggtttta aaacgaaaat tcttattctt gagtaactct
     1921 ttcctgtagg tcaggttgct ttctcaggta tagcatgagg tcgctcttat tgaccacacc
     1981 tctaccggca tgccgagcaa atgcctgcaa atcgctcccc atttcgctgg aaatctgctc
     2041 gtcagtggtg ctcacactga cgaatcatgt acagatcata ccgatgactg cctggcgact
     2101 cacaactaag caagacagcc ggaaccagcg ccggcgaaca ccactgcata tatggcatat
     2161 cacaacagtc cacgtctcaa gcagttacag agatgttacg aaccactagt gcactgcagt
     2221 acatgttata ccatagtagt agtttcaata atatattcca ctacttatat gtgttacccg
     2281 cattagaact cttattggtg gcgaaaatcg atggcaataa agaacggaag gggtttaata
     2341 gttgtatgct taacatattt cgatttaaat atataagaaa cgtcggtagc acaacaatta
     2401 actcattatt taggtatggc ggaaatacct gatgaaacca tccagcagtt catggcattg
     2461 accaatgtgt cgcataacat agccgttcaa tatctctctg aatttggaga tttaaatgaa
     2521 gcactaaatt cctattatgc ttctcaaacg gatgaccaaa aggatagaag agaggaagca
     2581 cattggaaca gacagcagga gaaggccctc aagcaagaag ccttctccac caactcttcg
     2641 aataaagcca taaatacgga gcacgttggt gggttatgtc caaaaccagg atcctcacaa
     2701 ggtagcaacg agtacttgaa aagccgagcg gccgcgtgtt acaaccaatt aaccaattct
     2761 gattagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca
     2821 ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc
     2881 cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa
     2941 cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg
     3001 actgaatccg gtgagaatgg caaaagctta tgcatttctt tccagacttg ttcaacaggc
     3061 cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat
     3121 tgcgcctgag cgaggcgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc
     3181 gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga
     3241 tattcttcta atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca
     3301 tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag
     3361 tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga
     3421 aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg
     3481 acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc
     3541 ggcctggagc aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt
     3601 atgtaagcag acagttttat tgttcatgat gatatatttt tatcttgtgc aatgtaacat
     3661 cagagatttt gagacacaac gtggctttgt tgaataaatc gaacttttgc tgagttgaag
     3721 gatcagtcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     3781 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     3841 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     3901 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     3961 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     4021 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     4081 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     4141 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     4201 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     4261 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     4321 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     4381 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     4441 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt gcggccgcca
     4501 attgtgaatg aagtagcgac agaaaaaaag cttttatttc gagttcgcat cattgtcact
     4561 ttcctgcttt ttcttggcgt actgtctatc taagatagtc tttaatatag cgtcctcgcc
     4621 atattcttct tcctcaattt tcttccatct cttttctaca tcagctctct ttattctact
     4681 ttcaatgaca gcacgtttct tgttattcaa acttgcctcc ttgagacact tgttcaaagc
     4741 atccttttcg ttgttacaaa ggccaaatat tctcttgtaa tattcctttt gatggcattt
     4801 atcgagtgca ttgatgaagt ccaaacaggc taaaatgaag aaggggagaa tacaatgtta
     4861 gtaagtcgga gcacagaaaa aggcaaatat atgaagtaaa atctggatac ttacaatgaa
     4921 aacgttcagc ttctaactgt ggatgcatgt tgtagtactt gtcttatctt gcttttgttc
     4981 aactgcactt gtaaatcagt gaccctgaat tcgcatctag actgatgaga cgtggtagag
     5041 ccacaaacag ccggtacaag caacgatctc caggaccatc tgaatcatgc gcggatgaca
     5101 cgaactcacg acggcgatca cagacattaa cccacagtac agacactgcg acaacgtggc
     5161 aattcgtcgc aatacaacg
//
