LOCUS       sequence-487894-       10293 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express Yea4p membrane protein cargo, peroxisomal Ynd1p truncation, 
            and Pex22 peroxisomal chauffeur; chromosomal integration by CRISPR .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10293)
  TITLE     Dueber Lab plasmids 2025
REFERENCE   2  (bases 1 to 10293)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 2, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pKS1604
FEATURES             Location/Qualifiers
     source          1..10293
                     /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             6444..7151
                     /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(8031..8846)
                     /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(8031..8846)
                     /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     8754..8773
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     8838..8857
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      9071..9659
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9560..9579
                     /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 tgccaccacc atcaagaagt agaataaaca
     6361 aaacaagaac attaggaata gtgggtacag ctatagcagt gttggtcacg tcctactata
     6421 tatatcaaaa ggtgacaggt tctgtgtcca aaggagagga gttaatcaag gaaaacatga
     6481 gaatgaaagt tgtcatggag ggctccgtta atggtcacca attcaagtgt acaggggaag
     6541 gtgaaggtaa tccttacatg ggtacacaaa ctatgagaat taaagtaatt gaaggcggac
     6601 cactaccatt tgcatttgac attctggcaa cgtcattcat gtacggatca cgaactttca
     6661 tcaagtaccc taaaggtata ccagactttt tcaagcaatc ttttccagag ggttttacat
     6721 gggaaagggt tacaagatac gaagatgggg gtgtcgtcac agttatgcaa gatacttcat
     6781 tagaagatgg ctgccttgtc tatcatgtgc aagtaagagg ggtgaatttt ccttctaacg
     6841 gacctgtgat gcagaaaaag accaaaggtt gggaaccaaa tactgaaatg atgtacccag
     6901 ctgatggagg tttgagaggc tacacacaca tggcgcttaa agttgatggt ggaggtcatt
     6961 tgtcttgtag ttttgttacc acttatcgtt ctaaaaagac tgttggcaat atcaaaatgc
     7021 caggaataca tgctgtagac cacagactag aaagactcga agagagcgat aacgaaatgt
     7081 tcgttgtaca gagagagcat gccgtagcca aatttgctgg cttaggcggt ggtatggatg
     7141 aattgtataa gggatccggg tcaggctccg gatctggccc tcccccagct cttccgccaa
     7201 aaagacgaag gtaatggcag tgcttttaac taagaattat tagtcttttc tgcttatttt
     7261 ttcatcatag tttagaacac tttatattaa cgaatagttt atgaatctat ttaggtttaa
     7321 aaattgatac agttttataa gttacttttt caaagactcg tgctgtctat tgcataatgc
     7381 actggaaggg gaaaaaaaag gtgcacacgc gtggcttttt cttgaatttg cagtttgaaa
     7441 aatgctgagc agttacagag atgttacgaa ccactagtgc actgcagtac atgacatgta
     7501 tgggttgaaa atatttagag gatgctaaaa gtcccgtcta aaatagagga agtcatggat
     7561 tacattatca cctttattgt cctgttccgt ctggctaatt tgggacgagg aacgtttgtc
     7621 tccattcgca accttttgaa accatatgaa cggcggctaa tctgacagat gtcttatatt
     7681 tctaggattg tagaaagcct cctcatcaaa ccaccttgtt tgaaaaaggc actagtgcgg
     7741 ttgctttact tgatctagta ttctcctacc ttggagcggt cccatatcaa ctaaattcag
     7801 ttttctgact gagttaaaaa ctaatgtagc gacaccactt ccattggtgc ctccgttatt
     7861 ttttttttgt atgttaatgc taaataatac accgctatgt atttcagggc actacttcta
     7921 cacatcaacg gtactaaaca tttcgctcaa gatcggtccg ctttcacgta tttggatgct
     7981 atgcaatgtt gccgagcggc cgcgtgttac aaccaattaa ccaattctga ttagaaaaac
     8041 tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt
     8101 tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca
     8161 agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc
     8221 ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt
     8281 gagaatggca aaagcttatg catttctttc cagacttgtt caacaggcca gccattacgc
     8341 tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg
     8401 aggcgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg
     8461 cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat
     8521 acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta
     8581 cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc
     8641 atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc
     8701 gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga
     8761 gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg cctggagcaa
     8821 gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac
     8881 agttttattg ttcatgatga tatattttta tcttgtgcaa tgtaacatca gagattttga
     8941 gacacaacgt ggctttgttg aataaatcga acttttgctg agttgaagga tcagtcatga
     9001 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     9061 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     9121 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     9181 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     9241 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     9301 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     9361 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     9421 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     9481 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     9541 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     9601 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     9661 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     9721 tctttcctgc gttatcccct gattctgtgg ataaccgtgc ggccgccaat tgaaaaaggc
     9781 cactactgag ccaagcgcag aaccagatga gccattcatt ggatacttgg gtgacgttac
     9841 tgcatctata aaccaagata taaaagaata tggtcgcaga agaacagtaa tacctacaag
     9901 cgatttagta gcgtcgttct acaccaaaaa gacgcctgtt tttaaggtca agttcagcag
     9961 aagcaattta gcccttgcag gaggtgcttt caggccataa ttcacactga aaaaaataac
    10021 agcatggaag tgatatgtaa taagtataac ctaatccgat atgtaactat tagatgcttt
    10081 ttcgacatcg tacttgacct gctcggcggt tatcatagcg tcttctttag cctttttttt
    10141 ttcgctcagt catcggggcc aaaacaagaa aggcgaagaa cagaaaaatg aaaaatgtaa
    10201 ctggttgttg taaaaacctt ccagagaaat aagatgaaat gtatgttgct ggcacgaacg
    10261 aagacaaata ccctgaattc gcatctagac tga
//
