LOCUS       sequence-432647-        8657 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express a red fluorescent protein to the peroxisome membrane via 
            tethering to a truncated Pex22. Also overexpresses Pex11.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8657)
  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 8657)
  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: pJJB1340
FEATURES             Location/Qualifiers
     source          1..8657
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        299..712
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      1458..1645
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1513..1537)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             2606..3313
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="VSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTMR
                     IKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGGV
                     VTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTHM
                     ALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAVA
                     KFAGLGGGMDELYK"
     promoter        3842..4249
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             4250..5344
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     CDS             complement(6375..7190)
                     /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(6375..7190)
                     /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     7098..7117
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     7182..7201
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      7415..8003
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7904..7923
                     /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 tatggtctgt gatacactgg tatatcatcc ctccgtgacg agattcgtca
      781 aatttctaga tggctcagct ggcagagaaa aggttctcag attactgcag tatttagcaa
      841 gatttttagc agtacagaac tcatctctct tggccaggca attacaagct caatttacca
      901 cagtcagaaa atttctgagg tttctgaagc ctttaaacca cttgcaggca gctgctaaat
      961 tctacgataa caagttggcc agcgacaacg tcgttagagt ctgcaatgtt ttgaaaaaca
     1021 tcttctttgc tgcatatttg tcgttagatc aagtcaatct tttgagaatt ttgaaagtga
     1081 ttcctgtaac cgttcttacc ggtaagaaaa tacctcgctg gtccaattgg tgttggctgt
     1141 tcggcctcct aagtggtctg gctatggatc ttcgtaagat ccaaacatct catgcccaga
     1201 ttgctgcgtt tgtcaaggca aaatcacaaa gccaaggcga tgagcatgag gatcacaaga
     1261 aggtactagg gaaggcatac caagacaggt acaccgcgtt aagaagacta ttctgggatg
     1321 ctgcagattc gttcatcgtc ctcaacaacc tagggtactt gtctagcaac gaagagtatg
     1381 ttgcgctatc cggtgttgtc acatctatcc ttggtatgca agacatgtgg aaagctacat
     1441 agggatccta actcgaggcg aatttcttat gatttatgat ttttattatt aaataagtta
     1501 taaaaaaaat aagtgtatac aaattttaaa gtgactctta ggttttaaaa cgaaaattct
     1561 tattcttgag taactctttc ctgtaggtca ggttgctttc tcaggtatag catgaggtcg
     1621 ctcttattga ccacacctct accggcatgc cgagcaaatg cctgcaaatc gctccccatt
     1681 tcgctgccaa accagatgtc aacacagcta caacgactag agatctatgt gaggatccta
     1741 actcgagatc gagctggatg cacacactgg cttaagatga caacgaagag gatgtccaat
     1801 atttttttta aggaataagg atacttcaag actagattcc cccctgcatt cccatcagaa
     1861 ccgtaaacct tggcgctttc cttgggaagt attcaagaag tgccttgtcc ggtttctgtg
     1921 gctcacaaac cagcgcgccc gatatggctt tcttttcact tatgaatgta ccagtacggg
     1981 acaattagaa cgctcctgta acaatctctt tgcaaatgtg gggttacatt ctaaccatgt
     2041 cacactgctg acgaaattca aagtaaaaaa aaatgggacc acgtcttgag aacgatagat
     2101 tttctttatt ttacattgaa cagtcgttgt ctcagcgcgc tttatgtttt cattcatact
     2161 tcatattata aaataacaaa agaagaattt catattcacg cccaagaaat caggctgctt
     2221 tccaaatgca attgacactt cattagccat cacacaaaac tctttcttgc tggagcttct
     2281 tttaaaaaag acctcagtac accaaacacg ttacccgacc tcgttatttt acgacaacta
     2341 tgataaaatt ctgaagaaaa aataaaaaaa ttttcatact tcttgctttt atttaaacca
     2401 ttgaatgatt tcttttgaac aaaactacct gtttcaccaa aggaaataga aagaaaaaat
     2461 caattagaag aaaacaaaaa acaaaagatc tatgccacca ccatcaagaa gtagaataaa
     2521 caaaacaaga acattaggaa tagtgggtac agctatagca gtgttggtca cgtcctacta
     2581 tatatatcaa aaggtgacag gttctgtgtc caaaggagag gagttaatca aggaaaacat
     2641 gagaatgaaa gttgtcatgg agggctccgt taatggtcac caattcaagt gtacagggga
     2701 aggtgaaggt aatccttaca tgggtacaca aactatgaga attaaagtaa ttgaaggcgg
     2761 accactacca tttgcatttg acattctggc aacgtcattc atgtacggat cacgaacttt
     2821 catcaagtac cctaaaggta taccagactt tttcaagcaa tcttttccag agggttttac
     2881 atgggaaagg gttacaagat acgaagatgg gggtgtcgtc acagttatgc aagatacttc
     2941 attagaagat ggctgccttg tctatcatgt gcaagtaaga ggggtgaatt ttccttctaa
     3001 cggacctgtg atgcagaaaa agaccaaagg ttgggaacca aatactgaaa tgatgtaccc
     3061 agctgatgga ggtttgagag gctacacaca catggcgctt aaagttgatg gtggaggtca
     3121 tttgtcttgt agttttgtta ccacttatcg ttctaaaaag actgttggca atatcaaaat
     3181 gccaggaata catgctgtag accacagact agaaagactc gaagagagcg ataacgaaat
     3241 gttcgttgta cagagagagc atgccgtagc caaatttgct ggcttaggcg gtggtatgga
     3301 tgaattgtat aagggatcct aactcgagag cttttgatta agccttctag tccaaaaaac
     3361 acgttttttt gtcatttatt tcattttctt agaatagttt agtttattca ttttatagtc
     3421 acgaatgttt tatgattcta tatagggttg caaacaagca tttttcattt tatgttaaaa
     3481 caatttcagg tttacctttt attctgcttg tggtgacgcg tgtatccgcc cgctcttttg
     3541 gtcacccatg tatgctgagc agttacagag atgttacgaa ccactagtgc actgcagtac
     3601 atcgaggaga acttctagta tatctacata cctaatatta ttgccttatt aaaaatggaa
     3661 tcccaacaat tacatcaaaa tccacattct cttcaaaatc aattgtcctg tacttccttg
     3721 ttcatgtgtg ttcaaaaacg ttatatttat aggataatta tactctattt ctcaacaagt
     3781 aattggttgt ttggccgagc ggtctaaggc gcctgattca agaaatatct tgaccgcagt
     3841 taactgtggg aatactcagg tatcgtaaga tgcaagagtt cgaatctctt agcaaccatt
     3901 atttttttcc tcaacataac gagaacacac aggggcgcta tcgcacagaa tcaaattcga
     3961 tgactggaaa ttttttgtta atttcagagg tcgcctgacg catatacctt tttcaactga
     4021 aaaattggga gaaaaaggaa aggtgagagc gccggaaccg gcttttcata tagaatagag
     4081 aagcgttcat gactaaatgc ttgcatcaca atacttgaag ttgacaatat tatttaagga
     4141 cctattgttt tttccaatag gtggttagca atcgtcttac tttctaactt ttcttacctt
     4201 ttacatttca gcaatatata tatatatatt tcaaggatat accattctaa tgtctgcccc
     4261 taagaagatc gtcgttttgc caggtgacca cgttggtcaa gaaatcacag ccgaagccat
     4321 taaggttctt aaagctattt ctgatgttcg ttccaatgtc aagttcgatt tcgaaaatca
     4381 tttaattggt ggtgctgcta tcgatgctac aggtgttcca cttccagatg aggcgctgga
     4441 agcctccaag aaggctgatg ccgttttgtt aggtgctgtg ggtggtccta aatggggaac
     4501 cggtagtgtt agacctgaac aaggtttact aaaaatccgt aaagaacttc aattgtacgc
     4561 caacttaaga ccatgtaact ttgcatccga ctctctttta gacttatctc caatcaagcc
     4621 acaatttgct aaaggtactg acttcgttgt tgtcagagaa ttagtgggag gtatttactt
     4681 tggtaagaga aaggaagacg atggtgatgg tgtcgcttgg gatagtgaac aatacaccgt
     4741 tccagaagtg caaagaatca caagaatggc cgctttcatg gccctacaac atgagccacc
     4801 attgcctatt tggtccttgg ataaagctaa tgttttggcc tcttcaagat tatggagaaa
     4861 aactgtggag gaaaccatca agaacgaatt tcctacattg aaggttcaac atcaattgat
     4921 tgattctgcc gccatgatcc tagttaagaa cccaacccac ctaaatggta ttataatcac
     4981 cagcaacatg tttggtgata tcatctccga tgaagcctcc gttatcccag gttccttggg
     5041 tttgttgcca tctgcgtcct tggcctcttt gccagacaag aacaccgcat ttggtttgta
     5101 cgaaccatgc cacggttctg ctccagattt gccaaagaat aaggtcaacc ctatcgccac
     5161 tatcttgtct gctgcaatga tgttgaaatt gtcattgaac ttgcctgaag aaggtaaggc
     5221 cattgaagat gcagttaaaa aggttttgga tgcaggtatc agaactggtg atttaggtgg
     5281 ttccaacagt accaccgaag tcggtgatgc tgtcgccgaa gaagttaaga aaatccttgc
     5341 ttaaaaagat tctctttttt tatgatattt gtacataaac tttataaatg aaattcataa
     5401 tagaaacgac acgaaattac aaaatggaat atgttcatag ggtagacgaa actatatacg
     5461 caatctacat acatttatca agaaggagaa aaaggaggat gtaaaggaat acaggtaagc
     5521 aaattgatac taatggctca acgtgataag gaaaaagaat tgcactttaa cattaatatt
     5581 gacaaggagg agggcaccac acaaaaagtt aggtgtaaca gaaaatcatg aaactatgat
     5641 tcctaattta tatattggag gattttctct aaaaaaaaaa aaatacaaca aataaaaaac
     5701 actcaatgac ctgaccattt gatggagttt aagtcaatac cttcttgaac catttcccat
     5761 aatggtgaaa gttccctcaa gaattttact ctgtcagaaa cggccttaac gacgtagtcg
     5821 agagtactcg tatcgcatgt cggtgcgaca cgaaattaca aaatggaata tgttcatagg
     5881 gtagacgaaa ctatatacgc aatctacata catttatcaa gaaggagaaa aaggaggatg
     5941 taaaggaata caggtaagca aattgatact aatggctcaa cgtgataagg aaaaagaatt
     6001 gcactttaac attaatattg acaaggagga gggcaccaca caaaaagtta ggtgtaacag
     6061 aaaatcatga aactatgatt cctaatttat atattggagg attttctcta aaaaaaaaaa
     6121 aatacaacaa ataaaaaaca ctcaatgacc tgaccatttg atggagttta agtcaatacc
     6181 ttcttgaacc atttcccata atggtgaaag ttccctcaag aattttactc tgtcagaaac
     6241 ggccttaacg acgtagtcga cctcctcttc agtactaaat ctaccaatac caaatctgat
     6301 ggaagaatgg gctaatgcat catccttacc cagcgccgag cggccgcgtg ttacaaccaa
     6361 ttaaccaatt ctgattagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata
     6421 tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca
     6481 ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca
     6541 acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca
     6601 ccatgagtga cgactgaatc cggtgagaat ggcaaaagct tatgcatttc tttccagact
     6661 tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta
     6721 ttcattcgtg attgcgcctg agcgaggcga aatacgcgat cgctgttaaa aggacaatta
     6781 caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca
     6841 cctgaatcag gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg
     6901 agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat
     6961 tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg
     7021 ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca
     7081 cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg
     7141 gaatttaatc gcggcctgga gcaagacgtt tcccgttgaa tatggctcat aacacccctt
     7201 gtattactgt ttatgtaagc agacagtttt attgttcatg atgatatatt tttatcttgt
     7261 gcaatgtaac atcagagatt ttgagacaca acgtggcttt gttgaataaa tcgaactttt
     7321 gctgagttga aggatcagtc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     7381 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     7441 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     7501 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     7561 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     7621 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     7681 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     7741 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     7801 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     7861 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     7921 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     7981 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     8041 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     8101 gtgcggccgc caatacccag aagaacagta aaataaagca aggtacgtga aattaatatt
     8161 tttaaatggt tctaaccgat gccgaagaac tgcgcagtcc ggttataacg tctgacatgt
     8221 ccttttttga tttggaatcc aaccactcaa gtgactctgt tcatttactt tgcgaaaaat
     8281 atacccacaa attgcccatc gaaagtgaat cgcaaaccac cttcagactg gcaccgacaa
     8341 agcaaagatt atacagacag agtactttat acgtaccgtt aagtctcaag caaagggttt
     8401 tcttatttac tgaacgggta aagagtatct gggccggctt gccaagatgc aaaccgaata
     8461 agtatttcaa agttgcattt gccttagccg tcctgacacc attggctatt tggatatttt
     8521 atattgactt tcgtgtacat tgatcacatc gactgttcta ttggcaaatg aaccacgggc
     8581 attgactatt tttcaggtta ctactatata ttatcatcac gggcaaggat tgtaccctga
     8641 attcgcatct agactga
//
