LOCUS       sequence-487896-       10668 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express Yea4p membrane protein cargo, peroxisomal Ynd1p truncation, 
            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 10668)
  TITLE     Dueber Lab plasmids 2025
REFERENCE   2  (bases 1 to 10668)
  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: pKS1606
FEATURES             Location/Qualifiers
     source          1..10668
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      5366..5553
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(5421..5445)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        5897..6310
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             6819..7526
                     /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(8406..9221)
                     /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(8406..9221)
                     /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     9129..9148
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     9213..9232
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      9446..10034
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9935..9954
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 actggccgat aattgcagac gaacgttgat aggtcaagat caatgtaaac aattactttg
       61 ttatgtagag tttttttagc tacctatatt ccaccataac atcaatcatg cggttgctgg
      121 tgtatttacc aataatgttt aatgtatata tatatatata tatatggggc cgtatactta
      181 catatagtag atgtcaagcg taggcgcttc ccctgccggc tgtgagggcg ccataaccaa
      241 ggtatctata gaccgccaat cagcaaacta cctccgtaca ttcatgttgc acccacacat
      301 ttatacaccc agaccgcgac aaattaccca taaggttgtt tgtgacggcg tcgtacaaga
      361 gaacgtggga actttttagg ctcaccaaaa aagaaagaaa aaatacgagt tgctgacaga
      421 agcctcaaga aaaaaaaaat tcttcttcga ctatgctgga ggcagagatg atcgagccgg
      481 tagttaacta tatatagcta aattggttcc atcaccttct tttctggtgt cgctccttct
      541 agtgctattt ctggcttttc ctattttttt ttttccattt ttctttctct ctttctaata
      601 tataaattct cttgcatttt ctatttttct ctctatctat tctacttgtt tattcccttc
      661 aaggtttttt tttaaggagt acttgttttt agaatatacg gtcaacgaac tataattaac
      721 taaacagatc tatgtggaac tcactaaaag cattcgctct agtgttcgga ggttgttgct
      781 ccaatgttat tacatttgaa accttaatga gcaacgaaac aggcagcatt aataatctca
      841 ttactttttg tcaattttta tttgttacat gtcagggact ccccgaattt ctggatgttc
      901 atcaaccttt cccttacttc aagcctctga aaactccact tcacgtttat gtcattactg
      961 tcgttttatt ttatatttca tctacgacga ataataacgt atttaaatac aacatatcca
     1021 ttccgattca tatcgttttc agatgcttcg gaaccgtaat aaccatgttt acatgctggt
     1081 tattaaatgg taggaaatat acaaagattc agattttatc aacactattt ttaaccattg
     1141 gagctataat tgcctcatta ttcaaagatg ctgattttcg gtaccaagat ttgaaactgc
     1201 aagcgtggaa aattggaagt gaccaatctg tagatctcac ttttattttt ggcatttgta
     1261 tactagtact ttcatcattc acatcatctt tactttcagc atacaatgaa cgtacgtatc
     1321 aaaaatatgg aaagcattgg aaggagaata tcttttacag tcatttttta tcattacctc
     1381 tctttctatt tagtcgaaag cagttaattc acgaatatcg agtgatgaga aagtctgaaa
     1441 gaatattgtg ttcaaatttt ggagggaaga ttctagtccc gcgagaagaa actctccttc
     1501 ttttcaatgt gttaacacaa tatttttgcg tcaagggtgt taatatcctc gccagtaaaa
     1561 caaacgctct gacactttcc ataacattac ttgtaagaaa atttataagt cttttactga
     1621 gcgtccgact tttcgataat aacttgtcat atactggcta catcggggtc tacctagtct
     1681 tttttggtgc ctttatatac tcgctgggat caattcaccc caggcaaaat gacaaggggg
     1741 cgataaagaa aagtaaaggt tctgtttcta aaggtgaaga attattcact ggtgttgtcc
     1801 caattttggt tgaattagat ggtgatgtta atggtcacaa attttctgtc tccggtgaag
     1861 gtgaaggtga tgctacttac ggtaaattga ccttaaaatt tatttgtact actggtaaat
     1921 tgccagttcc atggccaacc ttagtcacta ctttatcttg gggtgttcaa tgttttgcaa
     1981 gatacccaga tcatatgaaa caacatgact ttttcaagtc tgccatgcca gaaggttatg
     2041 ttcaagaaag aactattttt ttcaaagatg acggtaacta caagaccaga gctgaagtca
     2101 agtttgaagg tgatacctta gttaatagaa tcgaattaaa aggtattgat tttaaagaag
     2161 atggtaacat tttaggtcac aaattggaat acaattattt ctctgacaat gtttacatca
     2221 ctgctgacaa acaaaagaat ggtatcaaag ctaacttcaa aattagacac aacattgaag
     2281 atggtggtgt tcaattagct gaccattatc aacaaaatac tccaattggt gatggtccag
     2341 tcttgttacc agacaaccat tacttatcca ctcaatctaa gttatccaaa gatccaaacg
     2401 aaaagaggga ccacatggtc ttgttagaat ttgttactgc tgctggtatt accttgggta
     2461 tggatgaatt gtacaaagga tccgcagagt atgtgcgggc cctgtttgac tttaatggga
     2521 atgatgaaga agaccttccc tttaagaaag gagacatcct gagaatccgg gataagcctg
     2581 aagagcagtg gtggaatgca gaggacagcg aaggaaagag ggggatgatt cctgtccctt
     2641 acgtggagaa gtatggatct gggagcggat ctggaggcag catggcggaa tatgtacgag
     2701 ctctttttga ttttaatggc aatgatgagg aggatctccc ttttaaaaag ggcgatattc
     2761 tgcgcattcg tgataagcca gaggaacaat ggtggaacgc cgaggatagt gaggggaaac
     2821 gcggcatgat tccggtgccg tatgtagaga agtactaatg gcgccaattg gtgcggcaat
     2881 tgataataac gaaaatgtct tttaatgatc tgggtataat gaggaatttt ccgaacgttt
     2941 ttactttata tatatatata catgtaacat atattctata cgctatagag aaaggaaatt
     3001 tttcaattaa aaaaaaaata gagaaagagt ttcacttctt gattatcgct aacactaatg
     3061 gttgaagtac tgctacttta attttatgct gccaaaccag atgtcaacac agctacaacg
     3121 tatatattaa atttgctctt gttctgtact ttcctaattc ttatgtaaaa agacaagaat
     3181 ttatgatact atttaataac aaaaaactac ctaagaaaag catcatgcag tcgaaattga
     3241 aatcgaaaag taaaacttta acggaacatg tttgaaattc taagaaagca tacatcttca
     3301 tcccttatat atagagttat gtttgatatt agtagtcatg ttgtaatctc tggcctaagt
     3361 atacgtaacg aaaatggtag cacgtcgcgt ttatggcccc caggttaatg tgttctctga
     3421 aattcgcatc actttgagaa ataatgggaa caccttacgc gtgagctgtg cccaccgctt
     3481 cgcctaataa agcggtgttc tcaaaatttc tccccgtttt caggatcacg agcgccatct
     3541 agttctggta aaatcgcgct tacaagaaca aagaaaagaa acatcgcgta atgcaacagt
     3601 gagacacttg ccgtcatata taaggttttg gatcagtaac cgttatttga gcataacaca
     3661 ggtttttaaa tatattatta tatatcatgg tatatgtgta aaattttttt gctgactggt
     3721 tttgtttatt tatttagctt tttaaaaatt ttactttctt cttgttaatt ttttctgatt
     3781 gctctatact caaaccaaca acaacttact ctacaactaa gatctatgct catagaaaac
     3841 actaatgatc ggtttggtat cgtcatagat gcagggtctt cgggttcgag aatccatgtg
     3901 tttaagtggc aggacacgga atcgcttctt catgcaacaa accaggactc acagtccata
     3961 ttacaatcag tacctcatat tcatcaagaa aaagactgga cttttaagct gaatccaggc
     4021 ttgtcgagtt ttgaaaaaaa acctcaagat gcgtacaaat ctcacatcaa gccgttacta
     4081 gattttgcta agaatattat ccctgaatca cattggtcga gttgtcccgt ttttattcaa
     4141 gcaaccgcgg gcatgcgcct tttacctcaa gacatacaat cttccatttt ggatggtttg
     4201 tgtcaggggc tcaaacaccc tgcagaattt ttggttgagg attgctcagc acaaattcaa
     4261 gtcattgatg gtgaaaccga aggtttatat ggctggcttg gcttaaacta tctatatgga
     4321 cactttaatg actataatcc agaagtctct gaccatttta catttgggtt tatggacatg
     4381 ggcggtgcct ctactcagat tgcgtttgca ccgcatgatt caggagaaat agctagacat
     4441 agagatgaca tcgccaccat cttcttgagg agtgttaacg gagatttgca gaaatgggac
     4501 gtttttgtta gtacgtggtt agggtttggt gccaatcagg ctagaagaag gtacttggct
     4561 cagttgatca atacccttcc agaaaacaca aatgattacg aaaatgatga cttctctaca
     4621 aggaacttga atgatccatg tatgccgcga ggaagcagca cggattttga atttaaagat
     4681 accatatttc acatcgcggg ttctgggaat tacgaacaat gtacaaaatc tatttatccg
     4741 ttacttttaa agaacatgcc ttgtgatgac gagccgtgct tgtttaatgg tgttcatgct
     4801 cctcgaatag actttgccaa tgacaaattt ataggtactt ctgagtactg gtacactgcc
     4861 aacgacgtat tcaaacttgg gggcgaatac aactttgaca aatttagcaa aagcctaagg
     4921 gagttttgca attccaattg gacgcaaata ttagcgaaca gtgataaagg cgtgtataat
     4981 tctattccgg agaatttttt gaaagatgca tgctttaagg gcaattgggt ccttaatata
     5041 cttcatgaag ggtttgatat gcctcgaata gatgtcgacg cagaaaatgt caatgacagg
     5101 cccttatttc aaagtgtgga aaaagtcgaa gaacgagagt tatcgtggac actaggcaga
     5161 attttacttt atgcctcagg tagcatattg gcaggtaatg atgacttcat ggtgggtatt
     5221 gcgcccagtg aaagaagaac taagctcact ggtaagaaat tcatacctgg caagttactg
     5281 gagtctgatc aactacgcaa gcaaagttcc ggatccggtt cattgggaag aggtagaaga
     5341 tccaaattgt aataactcga gtggcgcgaa tttcttatga tttatgattt ttattattaa
     5401 ataagttata aaaaaaataa gtgtatacaa attttaaagt gactcttagg ttttaaaacg
     5461 aaaattctta ttcttgagta actctttcct gtaggtcagg ttgctttctc aggtatagca
     5521 tgaggtcgct cttattgacc acacctctac cggcatgccg agcaaatgcc tgcaaatcgc
     5581 tccccatttc gctggatgca cacactggct taagatgaca acgccttgcc aacagggagt
     5641 tcttcagaga catggaggct caaaacgaaa ttattgacag cctagacatc aatagtcata
     5701 caacagaaag cgaccaccca actttggctg ataatagcgt ataaacaatg catactttgt
     5761 acgttcaaaa tacaatgcag tagatatatt tatgcatatt acatataata catatcacat
     5821 aggaagcaac aggcgcgttg gacttttaat tttcgaggac cgcgaatcct tacatcacac
     5881 ccaatccccc acaagtgatc ccccacacac catagcttca aaatgtttct actccttttt
     5941 tactcttcca gattttctcg gactccgcgc atcgccgtac cacttcaaaa cacccaagca
     6001 cagcatacta aatttcccct ctttcttcct ctagggtgtc gttaattacc cgtactaaag
     6061 gtttggaaaa gaaaaaagac accgcctcgt ttctttttct tcgtcgaaaa aggcaataaa
     6121 aatttttatc acgtttcttt ttcttgaaaa tttttttttt tgattttttt ctctttcgat
     6181 gacctcccat tgatatttaa gttaataaac ggtcatcaat ttctcaagtt tcagtttcat
     6241 ttttcttgtt ctattacaac tttttttact tcttgctcat tagaaagaaa gcatagcaat
     6301 ctaatctaag ttttaattac aaaagatcta tgttaataga aagtttgata ggcgctgtta
     6361 ctggatttgc acaaatgctg gaatctactt atatggccac acacaactcg tttttcacca
     6421 tgatatccgt cgcagaacag tttggtaatt tgaaggaaat gctaggttca ttttttggta
     6481 tctttgctat aatgaagttt cttaaaaaaa tactctatcg tgctactaag gggagactag
     6541 gaataccacc aaagaatttc gcagaatctg aaggaagcaa aaataaacta attgaagatt
     6601 ttcaaaagtt taatgatagt ggtaccataa attctaatga gaaagccaca agaaggaaaa
     6661 tttcatggaa accgctacta ttttttttga tggcagtatt tggttttcca tatctactaa
     6721 acaaatttat tactaaacta cagacctctg gaaccatacg agcatcgcaa ggaaatggta
     6781 gcgagcctat tgatccttcg aagttagaat ttggttctgt gtccaaagga gaggagttaa
     6841 tcaaggaaaa catgagaatg aaagttgtca tggagggctc cgttaatggt caccaattca
     6901 agtgtacagg ggaaggtgaa ggtaatcctt acatgggtac acaaactatg agaattaaag
     6961 taattgaagg cggaccacta ccatttgcat ttgacattct ggcaacgtca ttcatgtacg
     7021 gatcacgaac tttcatcaag taccctaaag gtataccaga ctttttcaag caatcttttc
     7081 cagagggttt tacatgggaa agggttacaa gatacgaaga tgggggtgtc gtcacagtta
     7141 tgcaagatac ttcattagaa gatggctgcc ttgtctatca tgtgcaagta agaggggtga
     7201 attttccttc taacggacct gtgatgcaga aaaagaccaa aggttgggaa ccaaatactg
     7261 aaatgatgta cccagctgat ggaggtttga gaggctacac acacatggcg cttaaagttg
     7321 atggtggagg tcatttgtct tgtagttttg ttaccactta tcgttctaaa aagactgttg
     7381 gcaatatcaa aatgccagga atacatgctg tagaccacag actagaaaga ctcgaagaga
     7441 gcgataacga aatgttcgtt gtacagagag agcatgccgt agccaaattt gctggcttag
     7501 gcggtggtat ggatgaattg tataagggat ccgggtcagg ctccggatct ggccctcccg
     7561 cagctcttgc gccaaaaaga cgaaggtaat ggcagtgctt ttaactaaga attattagtc
     7621 ttttctgctt attttttcat catagtttag aacactttat attaacgaat agtttatgaa
     7681 tctatttagg tttaaaaatt gatacagttt tataagttac tttttcaaag actcgtgctg
     7741 tctattgcat aatgcactgg aaggggaaaa aaaaggtgca cacgcgtggc tttttcttga
     7801 atttgcagtt tgaaaaatgc tgagcagtta cagagatgtt acgaaccact agtgcactgc
     7861 agtacatgac atgtatgggt tgaaaatatt tagaggatgc taaaagtccc gtctaaaata
     7921 gaggaagtca tggattacat tatcaccttt attgtcctgt tccgtctggc taatttggga
     7981 cgaggaacgt ttgtctccat tcgcaacctt ttgaaaccat atgaacggcg gctaatctga
     8041 cagatgtctt atatttctag gattgtagaa agcctcctca tcaaaccacc ttgtttgaaa
     8101 aaggcactag tgcggttgct ttacttgatc tagtattctc ctaccttgga gcggtcccat
     8161 atcaactaaa ttcagttttc tgactgagtt aaaaactaat gtagcgacac cacttccatt
     8221 ggtgcctccg ttattttttt tttgtatgtt aatgctaaat aatacaccgc tatgtatttc
     8281 agggcactac ttctacacat caacggtact aaacatttcg ctcaagatcg gtccgctttc
     8341 acgtatttgg atgctatgca atgttgccga gcggccgcgt gttacaacca attaaccaat
     8401 tctgattaga aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta
     8461 tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag
     8521 ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata
     8581 caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg
     8641 acgactgaat ccggtgagaa tggcaaaagc ttatgcattt ctttccagac ttgttcaaca
     8701 ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt
     8761 gattgcgcct gagcgaggcg aaatacgcga tcgctgttaa aaggacaatt acaaacagga
     8821 atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca
     8881 ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat
     8941 gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc
     9001 cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc
     9061 agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc
     9121 ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat
     9181 cgcggcctgg agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg
     9241 tttatgtaag cagacagttt tattgttcat gatgatatat ttttatcttg tgcaatgtaa
     9301 catcagagat tttgagacac aacgtggctt tgttgaataa atcgaacttt tgctgagttg
     9361 aaggatcagt catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     9421 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     9481 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     9541 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     9601 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     9661 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     9721 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     9781 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     9841 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     9901 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     9961 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
    10021 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
    10081 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtgcggccg
    10141 ccaattgaaa aaggccacta ctgagccaag cgcagaacca gatgagccat tcattggata
    10201 cttgggtgac gttactgcat ctataaacca agatataaaa gaatatggtc gcagaagaac
    10261 agtaatacct acaagcgatt tagtagcgtc gttctacacc aaaaagacgc ctgtttttaa
    10321 ggtcaagttc agcagaagca atttagccct tgcaggaggt gctttcaggc cataattcac
    10381 actgaaaaaa ataacagcat ggaagtgata tgtaataagt ataacctaat ccgatatgta
    10441 actattagat gctttttcga catcgtactt gacctgctcg gcggttatca tagcgtcttc
    10501 tttagccttt tttttttcgc tcagtcatcg gggccaaaac aagaaaggcg aagaacagaa
    10561 aaatgaaaaa tgtaactggt tgttgtaaaa accttccaga gaaataagat gaaatgtatg
    10621 ttgctggcac gaacgaagac aaataccctg aattcgcatc tagactga
//
