LOCUS       sequence-479035-        7914 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express CpPT1(52-408) membrane enzyme cargo and inactive Pex22 
            peroxisomal chauffeur; chromosomal integration by CRISPR .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7914)
  TITLE     Dueber Lab plasmids 2025
REFERENCE   2  (bases 1 to 7914)
  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: pKS1603
FEATURES             Location/Qualifiers
     source          1..7914
                     /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             complement(5652..6467)
                     /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(5652..6467)
                     /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     6375..6394
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6459..6478
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6692..7280
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7181..7200
                     /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 catgacatgt atgggttgaa aatatttaga ggatgctaaa agtcccgtct
     5161 aaaatagagg aagtcatgga ttacattatc acctttattg tcctgttccg tctggctaat
     5221 ttgggacgag gaacgtttgt ctccattcgc aaccttttga aaccatatga acggcggcta
     5281 atctgacaga tgtcttatat ttctaggatt gtagaaagcc tcctcatcaa accaccttgt
     5341 ttgaaaaagg cactagtgcg gttgctttac ttgatctagt attctcctac cttggagcgg
     5401 tcccatatca actaaattca gttttctgac tgagttaaaa actaatgtag cgacaccact
     5461 tccattggtg cctccgttat tttttttttg tatgttaatg ctaaataata caccgctatg
     5521 tatttcaggg cactacttct acacatcaac ggtactaaac atttcgctca agatcggtcc
     5581 gctttcacgt atttggatgc tatgcaatgt tgccgagcgg ccgcgtgtta caaccaatta
     5641 accaattctg attagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca
     5701 ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg
     5761 aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca
     5821 tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca
     5881 tgagtgacga ctgaatccgg tgagaatggc aaaagcttat gcatttcttt ccagacttgt
     5941 tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc
     6001 attcgtgatt gcgcctgagc gaggcgaaat acgcgatcgc tgttaaaagg acaattacaa
     6061 acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct
     6121 gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc agtggtgagt
     6181 aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc
     6241 gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca
     6301 tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct
     6361 gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa
     6421 tttaatcgcg gcctggagca agacgtttcc cgttgaatat ggctcataac accccttgta
     6481 ttactgttta tgtaagcaga cagttttatt gttcatgatg atatattttt atcttgtgca
     6541 atgtaacatc agagattttg agacacaacg tggctttgtt gaataaatcg aacttttgct
     6601 gagttgaagg atcagtcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     6661 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     6721 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     6781 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     6841 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     6901 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     6961 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     7021 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     7081 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     7141 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     7201 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     7261 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     7321 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgtg
     7381 cggccgccaa ttgaaaaagg ccactactga gccaagcgca gaaccagatg agccattcat
     7441 tggatacttg ggtgacgtta ctgcatctat aaaccaagat ataaaagaat atggtcgcag
     7501 aagaacagta atacctacaa gcgatttagt agcgtcgttc tacaccaaaa agacgcctgt
     7561 ttttaaggtc aagttcagca gaagcaattt agcccttgca ggaggtgctt tcaggccata
     7621 attcacactg aaaaaaataa cagcatggaa gtgatatgta ataagtataa cctaatccga
     7681 tatgtaacta ttagatgctt tttcgacatc gtacttgacc tgctcggcgg ttatcatagc
     7741 gtcttcttta gccttttttt tttcgctcag tcatcggggc caaaacaaga aaggcgaaga
     7801 acagaaaaat gaaaaatgta actggttgtt gtaaaaacct tccagagaaa taagatgaaa
     7861 tgtatgttgc tggcacgaac gaagacaaat accctgaatt cgcatctaga ctga
//
