LOCUS       sequence-432601-        7037 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express a red fluorescent protein to the peroxisome membrane via 
            tethering to a truncated Pex22.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7037)
  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 7037)
  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: pJJB1338
FEATURES             Location/Qualifiers
     source          1..7037
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             986..1693
                     /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        2222..2629
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             2630..3724
                     /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(4755..5570)
                     /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(4755..5570)
                     /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     5478..5497
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5562..5581
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5795..6383
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6284..6303
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 actggccgat aattgcagac gaacgactag agatctatgt gaggatccta actcgagatc
       61 gagctgccaa accagatgtc aacacagcta caacgactag agatctatgt gaggatccta
      121 actcgagatc gagctggatg cacacactgg cttaagatga caacgaagag gatgtccaat
      181 atttttttta aggaataagg atacttcaag actagattcc cccctgcatt cccatcagaa
      241 ccgtaaacct tggcgctttc cttgggaagt attcaagaag tgccttgtcc ggtttctgtg
      301 gctcacaaac cagcgcgccc gatatggctt tcttttcact tatgaatgta ccagtacggg
      361 acaattagaa cgctcctgta acaatctctt tgcaaatgtg gggttacatt ctaaccatgt
      421 cacactgctg acgaaattca aagtaaaaaa aaatgggacc acgtcttgag aacgatagat
      481 tttctttatt ttacattgaa cagtcgttgt ctcagcgcgc tttatgtttt cattcatact
      541 tcatattata aaataacaaa agaagaattt catattcacg cccaagaaat caggctgctt
      601 tccaaatgca attgacactt cattagccat cacacaaaac tctttcttgc tggagcttct
      661 tttaaaaaag acctcagtac accaaacacg ttacccgacc tcgttatttt acgacaacta
      721 tgataaaatt ctgaagaaaa aataaaaaaa ttttcatact tcttgctttt atttaaacca
      781 ttgaatgatt tcttttgaac aaaactacct gtttcaccaa aggaaataga aagaaaaaat
      841 caattagaag aaaacaaaaa acaaaagatc tatgccacca ccatcaagaa gtagaataaa
      901 caaaacaaga acattaggaa tagtgggtac agctatagca gtgttggtca cgtcctacta
      961 tatatatcaa aaggtgacag gttctgtgtc caaaggagag gagttaatca aggaaaacat
     1021 gagaatgaaa gttgtcatgg agggctccgt taatggtcac caattcaagt gtacagggga
     1081 aggtgaaggt aatccttaca tgggtacaca aactatgaga attaaagtaa ttgaaggcgg
     1141 accactacca tttgcatttg acattctggc aacgtcattc atgtacggat cacgaacttt
     1201 catcaagtac cctaaaggta taccagactt tttcaagcaa tcttttccag agggttttac
     1261 atgggaaagg gttacaagat acgaagatgg gggtgtcgtc acagttatgc aagatacttc
     1321 attagaagat ggctgccttg tctatcatgt gcaagtaaga ggggtgaatt ttccttctaa
     1381 cggacctgtg atgcagaaaa agaccaaagg ttgggaacca aatactgaaa tgatgtaccc
     1441 agctgatgga ggtttgagag gctacacaca catggcgctt aaagttgatg gtggaggtca
     1501 tttgtcttgt agttttgtta ccacttatcg ttctaaaaag actgttggca atatcaaaat
     1561 gccaggaata catgctgtag accacagact agaaagactc gaagagagcg ataacgaaat
     1621 gttcgttgta cagagagagc atgccgtagc caaatttgct ggcttaggcg gtggtatgga
     1681 tgaattgtat aagggatcct aactcgagag cttttgatta agccttctag tccaaaaaac
     1741 acgttttttt gtcatttatt tcattttctt agaatagttt agtttattca ttttatagtc
     1801 acgaatgttt tatgattcta tatagggttg caaacaagca tttttcattt tatgttaaaa
     1861 caatttcagg tttacctttt attctgcttg tggtgacgcg tgtatccgcc cgctcttttg
     1921 gtcacccatg tatgctgagc agttacagag atgttacgaa ccactagtgc actgcagtac
     1981 atcgaggaga acttctagta tatctacata cctaatatta ttgccttatt aaaaatggaa
     2041 tcccaacaat tacatcaaaa tccacattct cttcaaaatc aattgtcctg tacttccttg
     2101 ttcatgtgtg ttcaaaaacg ttatatttat aggataatta tactctattt ctcaacaagt
     2161 aattggttgt ttggccgagc ggtctaaggc gcctgattca agaaatatct tgaccgcagt
     2221 taactgtggg aatactcagg tatcgtaaga tgcaagagtt cgaatctctt agcaaccatt
     2281 atttttttcc tcaacataac gagaacacac aggggcgcta tcgcacagaa tcaaattcga
     2341 tgactggaaa ttttttgtta atttcagagg tcgcctgacg catatacctt tttcaactga
     2401 aaaattggga gaaaaaggaa aggtgagagc gccggaaccg gcttttcata tagaatagag
     2461 aagcgttcat gactaaatgc ttgcatcaca atacttgaag ttgacaatat tatttaagga
     2521 cctattgttt tttccaatag gtggttagca atcgtcttac tttctaactt ttcttacctt
     2581 ttacatttca gcaatatata tatatatatt tcaaggatat accattctaa tgtctgcccc
     2641 taagaagatc gtcgttttgc caggtgacca cgttggtcaa gaaatcacag ccgaagccat
     2701 taaggttctt aaagctattt ctgatgttcg ttccaatgtc aagttcgatt tcgaaaatca
     2761 tttaattggt ggtgctgcta tcgatgctac aggtgttcca cttccagatg aggcgctgga
     2821 agcctccaag aaggctgatg ccgttttgtt aggtgctgtg ggtggtccta aatggggaac
     2881 cggtagtgtt agacctgaac aaggtttact aaaaatccgt aaagaacttc aattgtacgc
     2941 caacttaaga ccatgtaact ttgcatccga ctctctttta gacttatctc caatcaagcc
     3001 acaatttgct aaaggtactg acttcgttgt tgtcagagaa ttagtgggag gtatttactt
     3061 tggtaagaga aaggaagacg atggtgatgg tgtcgcttgg gatagtgaac aatacaccgt
     3121 tccagaagtg caaagaatca caagaatggc cgctttcatg gccctacaac atgagccacc
     3181 attgcctatt tggtccttgg ataaagctaa tgttttggcc tcttcaagat tatggagaaa
     3241 aactgtggag gaaaccatca agaacgaatt tcctacattg aaggttcaac atcaattgat
     3301 tgattctgcc gccatgatcc tagttaagaa cccaacccac ctaaatggta ttataatcac
     3361 cagcaacatg tttggtgata tcatctccga tgaagcctcc gttatcccag gttccttggg
     3421 tttgttgcca tctgcgtcct tggcctcttt gccagacaag aacaccgcat ttggtttgta
     3481 cgaaccatgc cacggttctg ctccagattt gccaaagaat aaggtcaacc ctatcgccac
     3541 tatcttgtct gctgcaatga tgttgaaatt gtcattgaac ttgcctgaag aaggtaaggc
     3601 cattgaagat gcagttaaaa aggttttgga tgcaggtatc agaactggtg atttaggtgg
     3661 ttccaacagt accaccgaag tcggtgatgc tgtcgccgaa gaagttaaga aaatccttgc
     3721 ttaaaaagat tctctttttt tatgatattt gtacataaac tttataaatg aaattcataa
     3781 tagaaacgac acgaaattac aaaatggaat atgttcatag ggtagacgaa actatatacg
     3841 caatctacat acatttatca agaaggagaa aaaggaggat gtaaaggaat acaggtaagc
     3901 aaattgatac taatggctca acgtgataag gaaaaagaat tgcactttaa cattaatatt
     3961 gacaaggagg agggcaccac acaaaaagtt aggtgtaaca gaaaatcatg aaactatgat
     4021 tcctaattta tatattggag gattttctct aaaaaaaaaa aaatacaaca aataaaaaac
     4081 actcaatgac ctgaccattt gatggagttt aagtcaatac cttcttgaac catttcccat
     4141 aatggtgaaa gttccctcaa gaattttact ctgtcagaaa cggccttaac gacgtagtcg
     4201 agagtactcg tatcgcatgt cggtgcgaca cgaaattaca aaatggaata tgttcatagg
     4261 gtagacgaaa ctatatacgc aatctacata catttatcaa gaaggagaaa aaggaggatg
     4321 taaaggaata caggtaagca aattgatact aatggctcaa cgtgataagg aaaaagaatt
     4381 gcactttaac attaatattg acaaggagga gggcaccaca caaaaagtta ggtgtaacag
     4441 aaaatcatga aactatgatt cctaatttat atattggagg attttctcta aaaaaaaaaa
     4501 aatacaacaa ataaaaaaca ctcaatgacc tgaccatttg atggagttta agtcaatacc
     4561 ttcttgaacc atttcccata atggtgaaag ttccctcaag aattttactc tgtcagaaac
     4621 ggccttaacg acgtagtcga cctcctcttc agtactaaat ctaccaatac caaatctgat
     4681 ggaagaatgg gctaatgcat catccttacc cagcgccgag cggccgcgtg ttacaaccaa
     4741 ttaaccaatt ctgattagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata
     4801 tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca
     4861 ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca
     4921 acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca
     4981 ccatgagtga cgactgaatc cggtgagaat ggcaaaagct tatgcatttc tttccagact
     5041 tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta
     5101 ttcattcgtg attgcgcctg agcgaggcga aatacgcgat cgctgttaaa aggacaatta
     5161 caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca
     5221 cctgaatcag gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg
     5281 agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat
     5341 tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg
     5401 ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca
     5461 cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg
     5521 gaatttaatc gcggcctgga gcaagacgtt tcccgttgaa tatggctcat aacacccctt
     5581 gtattactgt ttatgtaagc agacagtttt attgttcatg atgatatatt tttatcttgt
     5641 gcaatgtaac atcagagatt ttgagacaca acgtggcttt gttgaataaa tcgaactttt
     5701 gctgagttga aggatcagtc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5761 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5821 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5881 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     5941 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6001 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6061 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6121 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6181 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6241 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6301 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6361 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     6421 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     6481 gtgcggccgc caatacccag aagaacagta aaataaagca aggtacgtga aattaatatt
     6541 tttaaatggt tctaaccgat gccgaagaac tgcgcagtcc ggttataacg tctgacatgt
     6601 ccttttttga tttggaatcc aaccactcaa gtgactctgt tcatttactt tgcgaaaaat
     6661 atacccacaa attgcccatc gaaagtgaat cgcaaaccac cttcagactg gcaccgacaa
     6721 agcaaagatt atacagacag agtactttat acgtaccgtt aagtctcaag caaagggttt
     6781 tcttatttac tgaacgggta aagagtatct gggccggctt gccaagatgc aaaccgaata
     6841 agtatttcaa agttgcattt gccttagccg tcctgacacc attggctatt tggatatttt
     6901 atattgactt tcgtgtacat tgatcacatc gactgttcta ttggcaaatg aaccacgggc
     6961 attgactatt tttcaggtta ctactatata ttatcatcac gggcaaggat tgtaccctga
     7021 attcgcatct agactga
//
