LOCUS       sequence-475888-        9059 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express ClPT1(37-407) membrane enzyme cargo and Pex22 peroxisomal 
            chauffeur.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9059)
  TITLE     Dueber Lab plasmids 2025
REFERENCE   2  (bases 1 to 9059)
  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"
COMMENT     Sequence Label: pKS1517
FEATURES             Location/Qualifiers
     source          1..9059
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3560..3973
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             4107..4814
                     /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"
     CDS             5332..6135
                     /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(6777..7592)
                     /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(6777..7592)
                     /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     7500..7519
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     7584..7603
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      7817..8405
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8306..8325
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 actggccgat aattgcagac gaacgactag agatctatgt gaggatccta actcgagatc
       61 gagctgccaa accagatgtc aacacagcta caacgttgat aggtcaagat caatgtaaac
      121 aattactttg ttatgtagag tttttttagc tacctatatt ccaccataac atcaatcatg
      181 cggttgctgg tgtatttacc aataatgttt aatgtatata tatatatata tatatggggc
      241 cgtatactta catatagtag atgtcaagcg taggcgcttc ccctgccggc tgtgagggcg
      301 ccataaccaa ggtatctata gaccgccaat cagcaaacta cctccgtaca ttcatgttgc
      361 acccacacat ttatacaccc agaccgcgac aaattaccca taaggttgtt tgtgacggcg
      421 tcgtacaaga gaacgtggga actttttagg ctcaccaaaa aagaaagaaa aaatacgagt
      481 tgctgacaga agcctcaaga aaaaaaaaat tcttcttcga ctatgctgga ggcagagatg
      541 atcgagccgg tagttaacta tatatagcta aattggttcc atcaccttct tttctggtgt
      601 cgctccttct agtgctattt ctggcttttc ctattttttt ttttccattt ttctttctct
      661 ctttctaata tataaattct cttgcatttt ctatttttct ctctatctat tctacttgtt
      721 tattcccttc aaggtttttt tttaaggagt acttgttttt agaatatacg gtcaacgaac
      781 tataattaac taaacagatc tatggcagag tatgtgcggg ccctgtttga ctttaatggg
      841 aatgatgaag aagaccttcc ctttaagaaa ggagacatcc tgagaatccg ggataagcct
      901 gaagagcagt ggtggaatgc agaggacagc gaaggaaaga gggggatgat tcctgtccct
      961 tacgtggaga agtatggatc tgggagcgga tctggaggca gcatggcgga atatgtacga
     1021 gctctttttg attttaatgg caatgatgag gaggatctcc cttttaaaaa gggcgatatt
     1081 ctgcgcattc gtgataagcc agaggaacaa tggtggaacg ccgaggatag tgaggggaaa
     1141 cgcggcatga ttccggtgcc gtatgtagag aagtacggat ctggttctgt ttctaaaggt
     1201 gaagaattat tcactggtgt tgtcccaatt ttggttgaat tagatggtga tgttaatggt
     1261 cacaaatttt ctgtctccgg tgaaggtgaa ggtgatgcta cttacggtaa attgacctta
     1321 aaatttattt gtactactgg taaattgcca gttccatggc caaccttagt cactacttta
     1381 tcttggggtg ttcaatgttt tgcaagatac ccagatcata tgaaacaaca tgactttttc
     1441 aagtctgcca tgccagaagg ttatgttcaa gaaagaacta tttttttcaa agatgacggt
     1501 aactacaaga ccagagctga agtcaagttt gaaggtgata ccttagttaa tagaatcgaa
     1561 ttaaaaggta ttgattttaa agaagatggt aacattttag gtcacaaatt ggaatacaat
     1621 tatttctctg acaatgttta catcactgct gacaaacaaa agaatggtat caaagctaac
     1681 ttcaaaatta gacacaacat tgaagatggt ggtgttcaat tagctgacca ttatcaacaa
     1741 aatactccaa ttggtgatgg tccagtcttg ttaccagaca accattactt atccactcaa
     1801 tctaagttat ccaaagatcc aaacgaaaag agggaccaca tggtcttgtt agaatttgtt
     1861 actgctgctg gtattacctt gggtatggat gaattgtaca aaggatccgg tttaaacagg
     1921 tcagagtcaa agaactatgc aataaaatgt acccaatctg attcattcta ttcaaccaac
     1981 aagatcagaa ataacgaaaa ctcttcatca aggaattgta aaccatttaa taaatatagg
     2041 gttgccgtaa cattacaaca acaagactgt gcatcaaata atgaagatga tataaattca
     2101 acctctttta gggatgtatt attgaagaaa ttgcatgcct tatatgtctt tactagacct
     2161 tttgctatga taggaactat tgttggtatt acctctatcg ccattttacc attgcagtca
     2221 ttcgccgact tgacaccaaa gtattttatg gaatttctaa aggcattatt gtctgccgtt
     2281 ttgatgaata attatgtagg tacagttaac caggttgcag atgttgaaat tgataaggtt
     2341 aataaaccag gattaccttt agcatcaggt gatttatctg taggaacagg tttagcaata
     2401 acattgattt tgtctttgac atctttagca atagcattat cattgcaatc acctccttta
     2461 atctttggtt tgatagtatg gttcttgtta ggtacagcat actctgttga cttgccattc
     2521 ttgagatgga agactaaccc attcttagcc ggtatgtgta tggtaattgt ctttggatta
     2581 gtttatcaat tctctttctt catccacttc cagaaatatg ttttaggtag acctgttgtc
     2641 attacaagac ctttaatctt tgctgctgcc atcatttcta ctatttctgc cgtcatgtct
     2701 ttgttgaaag acatacctga tgaagatgga gacaaacagt tcggttatca gtctatttca
     2761 tctaagttag gtaaggagaa cgttttgagg ttgtgtgtct atgctttatt ctttgcctat
     2821 ggagtcgctg ttattgtagg tgcttcttct tcttttcaat tgggtaaatt agtttcaatt
     2881 attggacact ctacattggc cttcttgtta tggttaaggg ctcagactgt tgatttgtca
     2941 aataacgcat ctacattttc tttctactta ttcgtatgga agttattcta cggtgaatat
     3001 ttgttaatac acttcttgag ataatggcag tgcttttaac taagaattat tagtcttttc
     3061 tgcttatttt ttcatcatag tttagaacac tttatattaa cgaatagttt atgaatctat
     3121 ttaggtttaa aaattgatac agttttataa gttacttttt caaagactcg tgctgtctat
     3181 tgcataatgc actggaaggg gaaaaaaaag gtgcacacgc gtggcttttt cttgaatttg
     3241 cagtttgaaa aatgctggat gcacacactg gcttaagatg acaacgcctt gccaacaggg
     3301 agttcttcag agacatggag gctcaaaacg aaattattga cagcctagac atcaatagtc
     3361 atacaacaga aagcgaccac ccaactttgg ctgataatag cgtataaaca atgcatactt
     3421 tgtacgttca aaatacaatg cagtagatat atttatgcat attacatata atacatatca
     3481 cataggaagc aacaggcgcg ttggactttt aattttcgag gaccgcgaat ccttacatca
     3541 cacccaatcc cccacaagtg atcccccaca caccatagct tcaaaatgtt tctactcctt
     3601 ttttactctt ccagattttc tcggactccg cgcatcgccg taccacttca aaacacccaa
     3661 gcacagcata ctaaatttcc cctctttctt cctctagggt gtcgttaatt acccgtacta
     3721 aaggtttgga aaagaaaaaa gacaccgcct cgtttctttt tcttcgtcga aaaaggcaat
     3781 aaaaattttt atcacgtttc tttttcttga aaattttttt ttttgatttt tttctctttc
     3841 gatgacctcc cattgatatt taagttaata aacggtcatc aatttctcaa gtttcagttt
     3901 catttttctt gttctattac aacttttttt acttcttgct cattagaaag aaagcatagc
     3961 aatctaatct aagttttaat tacaaaagat ctatgccacc accatcaaga agtagaataa
     4021 acaaaacaag aacattagga atagtgggta cagctatagc agtgttggtc acgtcctact
     4081 atatatatca aaaggtgaca ggttctgtgt ccaaaggaga ggagttaatc aaggaaaaca
     4141 tgagaatgaa agttgtcatg gagggctccg ttaatggtca ccaattcaag tgtacagggg
     4201 aaggtgaagg taatccttac atgggtacac aaactatgag aattaaagta attgaaggcg
     4261 gaccactacc atttgcattt gacattctgg caacgtcatt catgtacgga tcacgaactt
     4321 tcatcaagta ccctaaaggt ataccagact ttttcaagca atcttttcca gagggtttta
     4381 catgggaaag ggttacaaga tacgaagatg ggggtgtcgt cacagttatg caagatactt
     4441 cattagaaga tggctgcctt gtctatcatg tgcaagtaag aggggtgaat tttccttcta
     4501 acggacctgt gatgcagaaa aagaccaaag gttgggaacc aaatactgaa atgatgtacc
     4561 cagctgatgg aggtttgaga ggctacacac acatggcgct taaagttgat ggtggaggtc
     4621 atttgtcttg tagttttgtt accacttatc gttctaaaaa gactgttggc aatatcaaaa
     4681 tgccaggaat acatgctgta gaccacagac tagaaagact cgaagagagc gataacgaaa
     4741 tgttcgttgt acagagagag catgccgtag ccaaatttgc tggcttaggc ggtggtatgg
     4801 atgaattgta taagggatcc gggtcaggct ccggatctgg ccctccccca gctcttccgc
     4861 caaaaagacg aaggtaatgg cagtgctttt aactaagaat tattagtctt ttctgcttat
     4921 tttttcatca tagtttagaa cactttatat taacgaatag tttatgaatc tatttaggtt
     4981 taaaaattga tacagtttta taagttactt tttcaaagac tcgtgctgtc tattgcataa
     5041 tgcactggaa ggggaaaaaa aaggtgcaca cgcgtggctt tttcttgaat ttgcagtttg
     5101 aaaaatgctg agcagttaca gagatgttac gaaccactag tgcactgcag tacacggttt
     5161 ccttgaaatt tttttgattc ggtaatctcc gaacagaagg aagaacgaag gaaggagcac
     5221 agacttagat tggtatatat acgcatatgt agtgttgaag aaacatgaaa ttgcccagta
     5281 ttcttaaccc aactgcacag aacaaaaacg tgcaggaaac gaagataaat catgtcgaaa
     5341 gctacatata aggaacgtgc tgctactcat cctagtcctg ttgctgccaa gctatttaat
     5401 atcatgcacg aaaagcaaac aaacttgtgt gcttcattgg atgttcgtac caccaaggaa
     5461 ttactggagt tagttgaagc attaggtccc aaaatttgtt tactaaaaac acatgtggat
     5521 atcttgactg atttttccat ggagggcaca gttaagccgc taaaggcatt atccgccaag
     5581 tacaattttt tactcttcga agacagaaaa tttgctgaca ttggtaatac agtcaaattg
     5641 cagtactctg cgggtgtata cagaatagca gaatgggcag acattacgaa tgcacacggt
     5701 gtggtgggcc caggtattgt tagcggtttg aagcaggcgg cagaagaagt aacaaaggaa
     5761 cctagaggcc ttttgatgtt agcagaattg tcatgcaagg gctccctatc tactggagaa
     5821 tatactaagg gtactgttga cattgcgaag agcgacaaag attttgttat cggctttatt
     5881 gctcaaagag acatgggtgg aagagatgaa ggttacgatt ggttgattat gacacccggt
     5941 gtgggtttag atgacaaggg agatgcattg ggtcaacagt atagaaccgt ggatgatgtg
     6001 gtttctacag gatctgacat tattattgtt ggaagaggac tatttgcaaa gggaagggat
     6061 gctaaggtag agggtgaacg ttacagaaaa gcaggctggg aagcatattt gagaagatgc
     6121 ggccagcaaa actaaaaaac tgtattataa gtaaatgcat gtatactaaa ctcacaaatt
     6181 agagcttcaa tttaattata tcagttatta cccgagtaga gcacttgaat ccactgcccc
     6241 gggaatctcg gtcgtaatga tttctataat gacgaaaaaa aaaaaattgg aaagaaaaag
     6301 cttcatggcc tttataaaaa ggaactatcc aatacctcgc cagaaccaag taacagtatt
     6361 ttacggggca caaatcaaga acaataagac aggactgtaa agatggacgc attgaactcc
     6421 aaagaacaac aagagttcca aaaagtagtg gaacaaaagc aaatgaagga tttcatgcgt
     6481 ttgtactcta atctggtaga aagatgtttc acagactgtg tcaatgactt cacaacatca
     6541 aagctaacca ataaggaaca aacatgcatc atgaagtgct cagaaaagtt cttgaagcat
     6601 agcgaacgtg tagggcagcg tttccaagaa caaaacgctg ccttgggaca aggcttgggc
     6661 cgataaggtg tactggcgta tatatatcta attatgtatc tctggtgtag cccattttta
     6721 gcatgtaaat ataaagaccg agcggccgcg tgttacaacc aattaaccaa ttctgattag
     6781 aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca
     6841 tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg
     6901 atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt
     6961 aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa
     7021 tccggtgaga atggcaaaag cttatgcatt tctttccaga cttgttcaac aggccagcca
     7081 ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc
     7141 tgagcgaggc gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc
     7201 aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct
     7261 tctaatacct ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca tgcatcatca
     7321 ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt
     7381 ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac
     7441 tctggcgcat cgggcttccc atacaatcga tagattgtcg cacctgattg cccgacatta
     7501 tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa tcgcggcctg
     7561 gagcaagacg tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa
     7621 gcagacagtt ttattgttca tgatgatata tttttatctt gtgcaatgta acatcagaga
     7681 ttttgagaca caacgtggct ttgttgaata aatcgaactt ttgctgagtt gaaggatcag
     7741 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     7801 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     7861 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     7921 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     7981 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     8041 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     8101 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     8161 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     8221 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     8281 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     8341 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     8401 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     8461 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtgcggcc gccaatgcta
     8521 aattcgagtg aaacacagga agatcagaaa atcctcattt catccatatt aacaataatt
     8581 tcaaatgttt atttgcatta tttgaaacta ggcaagacaa gcaacgaaac gtttttgaaa
     8641 attttgagta ttttcaataa atttgtagag gactcagata ttgaaaaaaa gctacagcaa
     8701 ttaatacttg ataagaagag tattgagaag ggcaacggtt catcatctca tggatctgca
     8761 catgaacaaa caccagagtc aaacgacgtt gaaattgagg ctactgcgcc aattgatgac
     8821 aatacagacg atgataacaa accgaagtta tctgatgtag aaaaggatta aagatgctaa
     8881 gagatagtga tgatatttca taaataatgt aattctatat atgttaatta ccttttttgc
     8941 gaggcatatt tatggtgaag gataagtttt gaccatcaaa gaaggttaat gtggctgtgg
     9001 tttcagggtc cataaagccc acatggataa cattacccct gaattcgcat ctagactga
//
