LOCUS       sequence-475863-        9674 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express CDT1 membrane protein cargo, peroxisomal Ynd1p truncation, 
            and inactive Pex22 peroxisomal chauffeur as negative control.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9674)
  TITLE     Dueber Lab plasmids 2025
REFERENCE   2  (bases 1 to 9674)
  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: pVL1281
FEATURES             Location/Qualifiers
     source          1..9674
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        4175..4588
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             4722..5429
                     /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             5947..6750
                     /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(7392..8207)
                     /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(7392..8207)
                     /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     8115..8134
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     8199..8218
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      8432..9020
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8921..8940
                     /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 tatgtcttct cacggctccc atgacggggc cagcaccgag aagcatcttg
      781 ctactcatga cattgcgccc acccacgacg ccatcaagat agtgcccaag ggccatggcc
      841 agacagccac aaagcccggt gcccaagaga aggaggtccg caacgccgcc ctatttgcgg
      901 ccatcaagga gtccaatatc aagccctgga gcaaggagtc catccacctc tatttcgcca
      961 tcttcgtcgc cttttgttgt gcatgcgcca acggttacga tggttcactc atgaccggaa
     1021 tcatcgctat ggacaagttc cagaaccaat tccacactgg tgacactggt cctaaagtct
     1081 ctgtcatctt ttctctctat accgttggtg ccatggttgg agctcccttc gctgctatcc
     1141 tctctgatcg ttttggccgt aagaagggca tgttcatcgg tggtatcttt atcattgtcg
     1201 gctccattat tgttgctagc tcctccaagc tcgctcagtt tgtcgttggc cgcttcgttc
     1261 ttggcctcgg tatcgccatc atgaccgttg ctgccccggc ctactccatc gaaatcgccc
     1321 ctcctcactg gcgcggtcgc tgcactggct tctactcctg cggttggttc ggaggttcga
     1381 ttcctgccgc ctgcatcacc tatggctgct acttcattaa gagcaactgg tcatggcgta
     1441 tccccttgat ccttcaggct ttcacgtgcc ttatcgtcat gtcctccgtc ttcttcctcc
     1501 cagaatcccc tcgctaccta tttgccaacg gccgcgacgc tgaggctgtt gcctttcttg
     1561 tcaagtatca cggcaacggc gatcccaatt ccaagctggt gttgctggag actgaggaga
     1621 tgagggacgg tatcaggacc gacggtgtcg acaaggtctg gtgggattac cgcccgctct
     1681 tcatgaccca cagcggtcgc tggcgcatgg cccaggtgct catgatctcc atctttggcc
     1741 agttctccgg caacggactc ggttacttca ataccgtcat cttcaagaac attggtgtca
     1801 ccagcacctc ccaacagctc gcctacaaca tcctcaactc cgtcatctcc gctatcggtg
     1861 ccttgaccgc cgtttccatg actgatcgta tgccccgccg cgcggtgctc attatcggta
     1921 ccttcatgtg cgccgctgct cttgccacca actcgggtct ttcggctact ctcgacaagc
     1981 agactcaaag aggcacgcaa atcaacctga accagggtat gaacgagcag gatgccaagg
     2041 acaacgccta cctccacgtc gacagcaact acgccaaggg tgccctggcc gcttacttcc
     2101 tcttcaacgt catcttctcc ttcacctaca ctcccctcca gggtgttatt cccaccgagg
     2161 ctctggaaac caccatccgt ggcaagggtc ttgccctttc cggcttcatt gtcaacgcca
     2221 tgggcttcat caaccagttc gctggcccca tcgctctcca caacattggc tacaagtaca
     2281 tctttgtctt tgtcggctgg gatcttatcg aaaccgtcgc ttggtacttc tttggtgtcg
     2341 aatcccaagg ccgtaccctg gagcagctcg aatgggtcta cgaccagccc aaccccgtca
     2401 aggcctccct aaaagtcgaa aaggtcgtcg tccaggccga cggccatgtg tccgaagcta
     2461 tcgttgctgg ttctgtttct aaaggtgaag aattattcac tggtgttgtc ccaattttgg
     2521 ttgaattaga tggtgatgtt aatggtcaca aattttctgt ctccggtgaa ggtgaaggtg
     2581 atgctactta cggtaaattg accttaaaat ttatttgtac tactggtaaa ttgccagttc
     2641 catggccaac cttagtcact actttatctt ggggtgttca atgttttgca agatacccag
     2701 atcatatgaa acaacatgac tttttcaagt ctgccatgcc agaaggttat gttcaagaaa
     2761 gaactatttt tttcaaagat gacggtaact acaagaccag agctgaagtc aagtttgaag
     2821 gtgatacctt agttaataga atcgaattaa aaggtattga ttttaaagaa gatggtaaca
     2881 ttttaggtca caaattggaa tacaattatt tctctgacaa tgtttacatc actgctgaca
     2941 aacaaaagaa tggtatcaaa gctaacttca aaattagaca caacattgaa gatggtggtg
     3001 ttcaattagc tgaccattat caacaaaata ctccaattgg tgatggtcca gtcttgttac
     3061 cagacaacca ttacttatcc actcaatcta agttatccaa agatccaaac gaaaagaggg
     3121 accacatggt cttgttagaa tttgttactg ctgctggtat taccttgggt atggatgaat
     3181 tgtacaaagg atccgcagag tatgtgcggg ccctgtttga ctttaatggg aatgatgaag
     3241 aagaccttcc ctttaagaaa ggagacatcc tgagaatccg ggataagcct gaagagcagt
     3301 ggtggaatgc agaggacagc gaaggaaaga gggggatgat tcctgtccct tacgtggaga
     3361 agtatggatc tgggagcgga tctggaggca gcatggcgga atatgtacga gctctttttg
     3421 attttaatgg caatgatgag gaggatctcc cttttaaaaa gggcgatatt ctgcgcattc
     3481 gtgataagcc agaggaacaa tggtggaacg ccgaggatag tgaggggaaa cgcggcatga
     3541 ttccggtgcc gtatgtagag aagtactaat ggcgccaatt ggtgcggcaa ttgataataa
     3601 cgaaaatgtc ttttaatgat ctgggtataa tgaggaattt tccgaacgtt tttactttat
     3661 atatatatat acatgtaaca tatattctat acgctataga gaaaggaaat ttttcaatta
     3721 aaaaaaaaat agagaaagag tttcacttct tgattatcgc taacactaat ggttgaagta
     3781 ctgctacttt aattttatgc tgccaaacca gatgtcaaca cagctacaac gactagagat
     3841 ctatgtgagg atcctaactc gagatcgagc tggatgcaca cactggctta agatgacaac
     3901 gccttgccaa cagggagttc ttcagagaca tggaggctca aaacgaaatt attgacagcc
     3961 tagacatcaa tagtcataca acagaaagcg accacccaac tttggctgat aatagcgtat
     4021 aaacaatgca tactttgtac gttcaaaata caatgcagta gatatattta tgcatattac
     4081 atataataca tatcacatag gaagcaacag gcgcgttgga cttttaattt tcgaggaccg
     4141 cgaatcctta catcacaccc aatcccccac aagtgatccc ccacacacca tagcttcaaa
     4201 atgtttctac tcctttttta ctcttccaga ttttctcgga ctccgcgcat cgccgtacca
     4261 cttcaaaaca cccaagcaca gcatactaaa tttcccctct ttcttcctct agggtgtcgt
     4321 taattacccg tactaaaggt ttggaaaaga aaaaagacac cgcctcgttt ctttttcttc
     4381 gtcgaaaaag gcaataaaaa tttttatcac gtttcttttt cttgaaaatt tttttttttg
     4441 atttttttct ctttcgatga cctcccattg atatttaagt taataaacgg tcatcaattt
     4501 ctcaagtttc agtttcattt ttcttgttct attacaactt tttttacttc ttgctcatta
     4561 gaaagaaagc atagcaatct aatctaagtt ttaattacaa aagatctatg ccaccaccat
     4621 caagaagtag aataaacaaa acaagaacat taggaatagt gggtacagct atagcagtgt
     4681 tggtcacgtc ctactatata tatcaaaagg tgacaggttc tgtgtccaaa ggagaggagt
     4741 taatcaagga aaacatgaga atgaaagttg tcatggaggg ctccgttaat ggtcaccaat
     4801 tcaagtgtac aggggaaggt gaaggtaatc cttacatggg tacacaaact atgagaatta
     4861 aagtaattga aggcggacca ctaccatttg catttgacat tctggcaacg tcattcatgt
     4921 acggatcacg aactttcatc aagtacccta aaggtatacc agactttttc aagcaatctt
     4981 ttccagaggg ttttacatgg gaaagggtta caagatacga agatgggggt gtcgtcacag
     5041 ttatgcaaga tacttcatta gaagatggct gccttgtcta tcatgtgcaa gtaagagggg
     5101 tgaattttcc ttctaacgga cctgtgatgc agaaaaagac caaaggttgg gaaccaaata
     5161 ctgaaatgat gtacccagct gatggaggtt tgagaggcta cacacacatg gcgcttaaag
     5221 ttgatggtgg aggtcatttg tcttgtagtt ttgttaccac ttatcgttct aaaaagactg
     5281 ttggcaatat caaaatgcca ggaatacatg ctgtagacca cagactagaa agactcgaag
     5341 agagcgataa cgaaatgttc gttgtacaga gagagcatgc cgtagccaaa tttgctggct
     5401 taggcggtgg tatggatgaa ttgtataagg gatccgggtc aggctccgga tctggccctc
     5461 ccgcagctct tgcgccaaaa agacgaaggt aatggcagtg cttttaacta agaattatta
     5521 gtcttttctg cttatttttt catcatagtt tagaacactt tatattaacg aatagtttat
     5581 gaatctattt aggtttaaaa attgatacag ttttataagt tactttttca aagactcgtg
     5641 ctgtctattg cataatgcac tggaagggga aaaaaaaggt gcacacgcgt ggctttttct
     5701 tgaatttgca gtttgaaaaa tgctgagcag ttacagagat gttacgaacc actagtgcac
     5761 tgcagtacac ggtttccttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa
     5821 cgaaggaagg agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca
     5881 tgaaattgcc cagtattctt aacccaactg cacagaacaa aaacgtgcag gaaacgaaga
     5941 taaatcatgt cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct
     6001 gccaagctat ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt
     6061 cgtaccacca aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta
     6121 aaaacacatg tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag
     6181 gcattatccg ccaagtacaa ttttttactc ttcgaagaca gaaaatttgc tgacattggt
     6241 aatacagtca aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt
     6301 acgaatgcac acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa
     6361 gaagtaacaa aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc
     6421 ctatctactg gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt
     6481 gttatcggct ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg
     6541 attatgacac ccggtgtggg tttagatgac aagggagatg cattgggtca acagtataga
     6601 accgtggatg atgtggtttc tacaggatct gacattatta ttgttggaag aggactattt
     6661 gcaaagggaa gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca
     6721 tatttgagaa gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata
     6781 ctaaactcac aaattagagc ttcaatttaa ttatatcagt tattacccga gtagagcact
     6841 tgaatccact gccccgggaa tctcggtcgt aatgatttct ataatgacga aaaaaaaaaa
     6901 attggaaaga aaaagcttca tggcctttat aaaaaggaac tatccaatac ctcgccagaa
     6961 ccaagtaaca gtattttacg gggcacaaat caagaacaat aagacaggac tgtaaagatg
     7021 gacgcattga actccaaaga acaacaagag ttccaaaaag tagtggaaca aaagcaaatg
     7081 aaggatttca tgcgtttgta ctctaatctg gtagaaagat gtttcacaga ctgtgtcaat
     7141 gacttcacaa catcaaagct aaccaataag gaacaaacat gcatcatgaa gtgctcagaa
     7201 aagttcttga agcatagcga acgtgtaggg cagcgtttcc aagaacaaaa cgctgccttg
     7261 ggacaaggct tgggccgata aggtgtactg gcgtatatat atctaattat gtatctctgg
     7321 tgtagcccat ttttagcatg taaatataaa gaccgagcgg ccgcgtgtta caaccaatta
     7381 accaattctg attagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca
     7441 ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg
     7501 aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca
     7561 tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca
     7621 tgagtgacga ctgaatccgg tgagaatggc aaaagcttat gcatttcttt ccagacttgt
     7681 tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc
     7741 attcgtgatt gcgcctgagc gaggcgaaat acgcgatcgc tgttaaaagg acaattacaa
     7801 acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct
     7861 gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc agtggtgagt
     7921 aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc
     7981 gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca
     8041 tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct
     8101 gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa
     8161 tttaatcgcg gcctggagca agacgtttcc cgttgaatat ggctcataac accccttgta
     8221 ttactgttta tgtaagcaga cagttttatt gttcatgatg atatattttt atcttgtgca
     8281 atgtaacatc agagattttg agacacaacg tggctttgtt gaataaatcg aacttttgct
     8341 gagttgaagg atcagtcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     8401 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     8461 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     8521 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     8581 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     8641 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     8701 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     8761 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     8821 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     8881 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     8941 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     9001 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     9061 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgtg
     9121 cggccgccaa tgctaaattc gagtgaaaca caggaagatc agaaaatcct catttcatcc
     9181 atattaacaa taatttcaaa tgtttatttg cattatttga aactaggcaa gacaagcaac
     9241 gaaacgtttt tgaaaatttt gagtattttc aataaatttg tagaggactc agatattgaa
     9301 aaaaagctac agcaattaat acttgataag aagagtattg agaagggcaa cggttcatca
     9361 tctcatggat ctgcacatga acaaacacca gagtcaaacg acgttgaaat tgaggctact
     9421 gcgccaattg atgacaatac agacgatgat aacaaaccga agttatctga tgtagaaaag
     9481 gattaaagat gctaagagat agtgatgata tttcataaat aatgtaattc tatatatgtt
     9541 aattaccttt tttgcgaggc atatttatgg tgaaggataa gttttgacca tcaaagaagg
     9601 ttaatgtggc tgtggtttca gggtccataa agcccacatg gataacatta cccctgaatt
     9661 cgcatctaga ctga
//
