LOCUS       sequence-475881-        9017 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express CpPT1(52-408) membrane enzyme cargo and inactive Pex22 
            peroxisomal chauffeur.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9017)
  TITLE     Dueber Lab plasmids 2025
REFERENCE   2  (bases 1 to 9017)
  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: pKS1520
FEATURES             Location/Qualifiers
     source          1..9017
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3518..3931
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             4065..4772
                     /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             5290..6093
                     /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(6735..7550)
                     /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(6735..7550)
                     /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     7458..7477
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     7542..7561
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      7775..8363
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8264..8283
                     /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 aaggatccat tcagtcttca
     1921 ttctgtttaa caaataataa gataggtaac aatgaagata tgatgaatag ataccataaa
     1981 ccattaaaga aatctacagt tcctatggct ttacaagatg actctgctac aaaatctcaa
     2041 aacgaaaata ttgtttctac atctttcttg gacttcttaa ccaagaagtt ggatgcattt
     2101 tataggttat caagaccata tgcttggact tcaatcattg taggtatttt gtcatcttct
     2161 ttgttgccaa tccaatcttt ggcagatttg actcctacct tcttgatcga ggtattgaag
     2221 cctatagttc cttcaattat gatgaatata tttgtcgttg ctattaatca gttatcagat
     2281 atagcaattg ataaggttaa caagccaaac ttgcctttag tttctggtga catatctatt
     2341 ggtgaggcta tagctattgc tgttatctca acattgactt ctttggctat gggtgttatg
     2401 ttaagatcac caccattagt tatagcatta atattgaggt gtatcttagg tgcagcttat
     2461 tctattgatt tgccattatt gagatggaag gcttctcctt tgatggctgc cgtcgcaatc
     2521 gttattggaa atggtataaa taatgttttg ccatacttct tacacgtcca aaagtacgtt
     2581 ttaggtaggc ctgttgtctt tactaagcct ttattattcg cagttgcatt ctcagcaata
     2641 ttttctatcg tcttatcatt cttaaaagac attcctgacg ttgaaggaga taagaaatct
     2701 ggtatcagaa cattaccagt cattttgggt aaagagagag tcttgtctat gtcaactggt
     2761 attttgttga tggcctatgc ttctgccgcc ttggctggag tcttctcacc aattttatta
     2821 tgtaaattgg taactatgat tggtcattca gtattgggtt ttatcttatg gtctaaggcc
     2881 caaacagttg atttatcaaa tgctaaatct acatattcat tcttcttatt cattttccag
     2941 ttgtattaca ccgagttctt cttaatgcat tttgtaagat aatggcagtg cttttaacta
     3001 agaattatta gtcttttctg cttatttttt catcatagtt tagaacactt tatattaacg
     3061 aatagtttat gaatctattt aggtttaaaa attgatacag ttttataagt tactttttca
     3121 aagactcgtg ctgtctattg cataatgcac tggaagggga aaaaaaaggt gcacacgcgt
     3181 ggctttttct tgaatttgca gtttgaaaaa tgctggatgc acacactggc ttaagatgac
     3241 aacgccttgc caacagggag ttcttcagag acatggaggc tcaaaacgaa attattgaca
     3301 gcctagacat caatagtcat acaacagaaa gcgaccaccc aactttggct gataatagcg
     3361 tataaacaat gcatactttg tacgttcaaa atacaatgca gtagatatat ttatgcatat
     3421 tacatataat acatatcaca taggaagcaa caggcgcgtt ggacttttaa ttttcgagga
     3481 ccgcgaatcc ttacatcaca cccaatcccc cacaagtgat cccccacaca ccatagcttc
     3541 aaaatgtttc tactcctttt ttactcttcc agattttctc ggactccgcg catcgccgta
     3601 ccacttcaaa acacccaagc acagcatact aaatttcccc tctttcttcc tctagggtgt
     3661 cgttaattac ccgtactaaa ggtttggaaa agaaaaaaga caccgcctcg tttctttttc
     3721 ttcgtcgaaa aaggcaataa aaatttttat cacgtttctt tttcttgaaa attttttttt
     3781 ttgatttttt tctctttcga tgacctccca ttgatattta agttaataaa cggtcatcaa
     3841 tttctcaagt ttcagtttca tttttcttgt tctattacaa ctttttttac ttcttgctca
     3901 ttagaaagaa agcatagcaa tctaatctaa gttttaatta caaaagatct atgccaccac
     3961 catcaagaag tagaataaac aaaacaagaa cattaggaat agtgggtaca gctatagcag
     4021 tgttggtcac gtcctactat atatatcaaa aggtgacagg ttctgtgtcc aaaggagagg
     4081 agttaatcaa ggaaaacatg agaatgaaag ttgtcatgga gggctccgtt aatggtcacc
     4141 aattcaagtg tacaggggaa ggtgaaggta atccttacat gggtacacaa actatgagaa
     4201 ttaaagtaat tgaaggcgga ccactaccat ttgcatttga cattctggca acgtcattca
     4261 tgtacggatc acgaactttc atcaagtacc ctaaaggtat accagacttt ttcaagcaat
     4321 cttttccaga gggttttaca tgggaaaggg ttacaagata cgaagatggg ggtgtcgtca
     4381 cagttatgca agatacttca ttagaagatg gctgccttgt ctatcatgtg caagtaagag
     4441 gggtgaattt tccttctaac ggacctgtga tgcagaaaaa gaccaaaggt tgggaaccaa
     4501 atactgaaat gatgtaccca gctgatggag gtttgagagg ctacacacac atggcgctta
     4561 aagttgatgg tggaggtcat ttgtcttgta gttttgttac cacttatcgt tctaaaaaga
     4621 ctgttggcaa tatcaaaatg ccaggaatac atgctgtaga ccacagacta gaaagactcg
     4681 aagagagcga taacgaaatg ttcgttgtac agagagagca tgccgtagcc aaatttgctg
     4741 gcttaggcgg tggtatggat gaattgtata agggatccgg gtcaggctcc ggatctggcc
     4801 ctcccgcagc tcttgcgcca aaaagacgaa ggtaatggca gtgcttttaa ctaagaatta
     4861 ttagtctttt ctgcttattt tttcatcata gtttagaaca ctttatatta acgaatagtt
     4921 tatgaatcta tttaggttta aaaattgata cagttttata agttactttt tcaaagactc
     4981 gtgctgtcta ttgcataatg cactggaagg ggaaaaaaaa ggtgcacacg cgtggctttt
     5041 tcttgaattt gcagtttgaa aaatgctgag cagttacaga gatgttacga accactagtg
     5101 cactgcagta cacggtttcc ttgaaatttt tttgattcgg taatctccga acagaaggaa
     5161 gaacgaagga aggagcacag acttagattg gtatatatac gcatatgtag tgttgaagaa
     5221 acatgaaatt gcccagtatt cttaacccaa ctgcacagaa caaaaacgtg caggaaacga
     5281 agataaatca tgtcgaaagc tacatataag gaacgtgctg ctactcatcc tagtcctgtt
     5341 gctgccaagc tatttaatat catgcacgaa aagcaaacaa acttgtgtgc ttcattggat
     5401 gttcgtacca ccaaggaatt actggagtta gttgaagcat taggtcccaa aatttgttta
     5461 ctaaaaacac atgtggatat cttgactgat ttttccatgg agggcacagt taagccgcta
     5521 aaggcattat ccgccaagta caatttttta ctcttcgaag acagaaaatt tgctgacatt
     5581 ggtaatacag tcaaattgca gtactctgcg ggtgtataca gaatagcaga atgggcagac
     5641 attacgaatg cacacggtgt ggtgggccca ggtattgtta gcggtttgaa gcaggcggca
     5701 gaagaagtaa caaaggaacc tagaggcctt ttgatgttag cagaattgtc atgcaagggc
     5761 tccctatcta ctggagaata tactaagggt actgttgaca ttgcgaagag cgacaaagat
     5821 tttgttatcg gctttattgc tcaaagagac atgggtggaa gagatgaagg ttacgattgg
     5881 ttgattatga cacccggtgt gggtttagat gacaagggag atgcattggg tcaacagtat
     5941 agaaccgtgg atgatgtggt ttctacagga tctgacatta ttattgttgg aagaggacta
     6001 tttgcaaagg gaagggatgc taaggtagag ggtgaacgtt acagaaaagc aggctgggaa
     6061 gcatatttga gaagatgcgg ccagcaaaac taaaaaactg tattataagt aaatgcatgt
     6121 atactaaact cacaaattag agcttcaatt taattatatc agttattacc cgagtagagc
     6181 acttgaatcc actgccccgg gaatctcggt cgtaatgatt tctataatga cgaaaaaaaa
     6241 aaaattggaa agaaaaagct tcatggcctt tataaaaagg aactatccaa tacctcgcca
     6301 gaaccaagta acagtatttt acggggcaca aatcaagaac aataagacag gactgtaaag
     6361 atggacgcat tgaactccaa agaacaacaa gagttccaaa aagtagtgga acaaaagcaa
     6421 atgaaggatt tcatgcgttt gtactctaat ctggtagaaa gatgtttcac agactgtgtc
     6481 aatgacttca caacatcaaa gctaaccaat aaggaacaaa catgcatcat gaagtgctca
     6541 gaaaagttct tgaagcatag cgaacgtgta gggcagcgtt tccaagaaca aaacgctgcc
     6601 ttgggacaag gcttgggccg ataaggtgta ctggcgtata tatatctaat tatgtatctc
     6661 tggtgtagcc catttttagc atgtaaatat aaagaccgag cggccgcgtg ttacaaccaa
     6721 ttaaccaatt ctgattagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata
     6781 tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca
     6841 ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca
     6901 acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca
     6961 ccatgagtga cgactgaatc cggtgagaat ggcaaaagct tatgcatttc tttccagact
     7021 tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta
     7081 ttcattcgtg attgcgcctg agcgaggcga aatacgcgat cgctgttaaa aggacaatta
     7141 caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca
     7201 cctgaatcag gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg
     7261 agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat
     7321 tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg
     7381 ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca
     7441 cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg
     7501 gaatttaatc gcggcctgga gcaagacgtt tcccgttgaa tatggctcat aacacccctt
     7561 gtattactgt ttatgtaagc agacagtttt attgttcatg atgatatatt tttatcttgt
     7621 gcaatgtaac atcagagatt ttgagacaca acgtggcttt gttgaataaa tcgaactttt
     7681 gctgagttga aggatcagtc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     7741 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     7801 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     7861 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     7921 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     7981 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     8041 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     8101 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     8161 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     8221 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     8281 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     8341 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     8401 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     8461 gtgcggccgc caatgctaaa ttcgagtgaa acacaggaag atcagaaaat cctcatttca
     8521 tccatattaa caataatttc aaatgtttat ttgcattatt tgaaactagg caagacaagc
     8581 aacgaaacgt ttttgaaaat tttgagtatt ttcaataaat ttgtagagga ctcagatatt
     8641 gaaaaaaagc tacagcaatt aatacttgat aagaagagta ttgagaaggg caacggttca
     8701 tcatctcatg gatctgcaca tgaacaaaca ccagagtcaa acgacgttga aattgaggct
     8761 actgcgccaa ttgatgacaa tacagacgat gataacaaac cgaagttatc tgatgtagaa
     8821 aaggattaaa gatgctaaga gatagtgatg atatttcata aataatgtaa ttctatatat
     8881 gttaattacc ttttttgcga ggcatattta tggtgaagga taagttttga ccatcaaaga
     8941 aggttaatgt ggctgtggtt tcagggtcca taaagcccac atggataaca ttacccctga
     9001 attcgcatct agactga
//
