LOCUS       sequence-475862-        9413 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express CDT1 membrane protein cargo fused directly to Pex22 TMD, 
            peroxisomal Ynd1p truncation, and peroxisomal mRuby2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9413)
  TITLE     Dueber Lab plasmids 2025
REFERENCE   2  (bases 1 to 9413)
  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: pVL1279
FEATURES             Location/Qualifiers
     source          1..9413
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3914..4327
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             4461..5168
                     /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             5686..6489
                     /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(7131..7946)
                     /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(7131..7946)
                     /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     7854..7873
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     7938..7957
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      8171..8759
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8660..8679
                     /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 tatgccacca ccatcaagaa gtagaataaa caaaacaaga acattaggaa
      781 tagtgggtac agctatagca gtgttggtca cgtcctacta tatatatcaa aaggtgacag
      841 gttcttcttc tcacggctcc catgacgggg ccagcaccga gaagcatctt gctactcatg
      901 acattgcgcc cacccacgac gccatcaaga tagtgcccaa gggccatggc cagacagcca
      961 caaagcccgg tgcccaagag aaggaggtcc gcaacgccgc cctatttgcg gccatcaagg
     1021 agtccaatat caagccctgg agcaaggagt ccatccacct ctatttcgcc atcttcgtcg
     1081 ccttttgttg tgcatgcgcc aacggttacg atggttcact catgaccgga atcatcgcta
     1141 tggacaagtt ccagaaccaa ttccacactg gtgacactgg tcctaaagtc tctgtcatct
     1201 tttctctcta taccgttggt gccatggttg gagctccctt cgctgctatc ctctctgatc
     1261 gttttggccg taagaagggc atgttcatcg gtggtatctt tatcattgtc ggctccatta
     1321 ttgttgctag ctcctccaag ctcgctcagt ttgtcgttgg ccgcttcgtt cttggcctcg
     1381 gtatcgccat catgaccgtt gctgccccgg cctactccat cgaaatcgcc cctcctcact
     1441 ggcgcggtcg ctgcactggc ttctacaact gcggttggtt cggaggttcg attcctgccg
     1501 cctgcatcac ctatggctgc tacttcatta agagcaactg gtcatggcgt atccccttga
     1561 tccttcaggc tttcacgtgc cttatcgtca tgtcctccgt cttcttcctc ccagaatccc
     1621 ctcgcttcct atttgccaac ggccgcgacg ctgaggctgt tgcctttctt gtcaagtatc
     1681 acggcaacgg cgatcccaat tccaagctgg tgttgctgga gactgaggag atgagggacg
     1741 gtatcaggac cgacggtgtc gacaaggtct ggtgggatta ccgcccgctc ttcatgaccc
     1801 acagcggtcg ctggcgcatg gcccaggtgc tcatgatctc catctttggc cagttctccg
     1861 gcaacggact cggttacttc aataccgtca tcttcaagaa cattggtgtc accagcacct
     1921 cccaacagct cgcctacaac atcctcaact ccgtcatctc cgctatcggt gccttgaccg
     1981 ccgtttccat gactgatcgt atgccccgcc gcgcggtgct cattatcggt accttcatgt
     2041 gcgccgctgc tcttgccacc aactcgggtc tttcggctac tctcgacaag cagactcaaa
     2101 gaggcacgca aatcaacctg aaccagggta tgaacgagca ggatgccaag gacaacgcct
     2161 acctccacgt cgacagcaac tacgccaagg gtgccctggc cgcttacttc ctcttcaacg
     2221 tcatcttctc cttcacctac actcccctcc agggtgttat tcccaccgag gctctggaaa
     2281 ccaccatccg tggcaagggt cttgcccttt ccggcttcat tgtcaacgcc atgggcttca
     2341 tcaaccagtt cgctggcccc atcgctctcc acaacattgg ctacaagtac atctttgtct
     2401 ttgtcggctg ggatcttatc gaaaccgtcg cttggtactt ctttggtgtc gaatcccaag
     2461 gccgtaccct ggagcagctc gaatgggtct acgaccagcc caaccccgtc aaggcctccc
     2521 taaaagtcga aaaggtcgtc gtccaggccg acggccatgt gtccgaagct atcgttgctg
     2581 gatccgtttc taaaggtgaa gaattattca ctggtgttgt cccaattttg gttgaattag
     2641 atggtgatgt taatggtcac aaattttctg tctccggtga aggtgaaggt gatgctactt
     2701 acggtaaatt gaccttaaaa tttatttgta ctactggtaa attgccagtt ccatggccaa
     2761 ccttagtcac tactttatct tggggtgttc aatgttttgc aagataccca gatcatatga
     2821 aacaacatga ctttttcaag tctgccatgc cagaaggtta tgttcaagaa agaactattt
     2881 ttttcaaaga tgacggtaac tacaagacca gagctgaagt caagtttgaa ggtgatacct
     2941 tagttaatag aatcgaatta aaaggtattg attttaaaga agatggtaac attttaggtc
     3001 acaaattgga atacaattat ttctctgaca atgtttacat cactgctgac aaacaaaaga
     3061 atggtatcaa agctaacttc aaaattagac acaacattga agatggtggt gttcaattag
     3121 ctgaccatta tcaacaaaat actccaattg gtgatggtcc agtcttgtta ccagacaacc
     3181 attacttatc cactcaatct aagttatcca aagatccaaa cgaaaagagg gaccacatgg
     3241 tcttgttaga atttgttact gctgctggta ttaccttggg tatggatgaa ttgtacaaat
     3301 aactcgagtg gcgccaattg gtgcggcaat tgataataac gaaaatgtct tttaatgatc
     3361 tgggtataat gaggaatttt ccgaacgttt ttactttata tatatatata catgtaacat
     3421 atattctata cgctatagag aaaggaaatt tttcaattaa aaaaaaaata gagaaagagt
     3481 ttcacttctt gattatcgct aacactaatg gttgaagtac tgctacttta attttatgct
     3541 gccaaaccag atgtcaacac agctacaacg actagagatc tatgtgagga tcctaactcg
     3601 agatcgagct ggatgcacac actggcttaa gatgacaacg ccttgccaac agggagttct
     3661 tcagagacat ggaggctcaa aacgaaatta ttgacagcct agacatcaat agtcatacaa
     3721 cagaaagcga ccacccaact ttggctgata atagcgtata aacaatgcat actttgtacg
     3781 ttcaaaatac aatgcagtag atatatttat gcatattaca tataatacat atcacatagg
     3841 aagcaacagg cgcgttggac ttttaatttt cgaggaccgc gaatccttac atcacaccca
     3901 atcccccaca agtgatcccc cacacaccat agcttcaaaa tgtttctact ccttttttac
     3961 tcttccagat tttctcggac tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag
     4021 catactaaat ttcccctctt tcttcctcta gggtgtcgtt aattacccgt actaaaggtt
     4081 tggaaaagaa aaaagacacc gcctcgtttc tttttcttcg tcgaaaaagg caataaaaat
     4141 ttttatcacg tttctttttc ttgaaaattt ttttttttga tttttttctc tttcgatgac
     4201 ctcccattga tatttaagtt aataaacggt catcaatttc tcaagtttca gtttcatttt
     4261 tcttgttcta ttacaacttt ttttacttct tgctcattag aaagaaagca tagcaatcta
     4321 atctaagttt taattacaaa agatctatgc caccaccatc aagaagtaga ataaacaaaa
     4381 caagaacatt aggaatagtg ggtacagcta tagcagtgtt ggtcacgtcc tactatatat
     4441 atcaaaaggt gacaggttct gtgtccaaag gagaggagtt aatcaaggaa aacatgagaa
     4501 tgaaagttgt catggagggc tccgttaatg gtcaccaatt caagtgtaca ggggaaggtg
     4561 aaggtaatcc ttacatgggt acacaaacta tgagaattaa agtaattgaa ggcggaccac
     4621 taccatttgc atttgacatt ctggcaacgt cattcatgta cggatcacga actttcatca
     4681 agtaccctaa aggtatacca gactttttca agcaatcttt tccagagggt tttacatggg
     4741 aaagggttac aagatacgaa gatgggggtg tcgtcacagt tatgcaagat acttcattag
     4801 aagatggctg ccttgtctat catgtgcaag taagaggggt gaattttcct tctaacggac
     4861 ctgtgatgca gaaaaagacc aaaggttggg aaccaaatac tgaaatgatg tacccagctg
     4921 atggaggttt gagaggctac acacacatgg cgcttaaagt tgatggtgga ggtcatttgt
     4981 cttgtagttt tgttaccact tatcgttcta aaaagactgt tggcaatatc aaaatgccag
     5041 gaatacatgc tgtagaccac agactagaaa gactcgaaga gagcgataac gaaatgttcg
     5101 ttgtacagag agagcatgcc gtagccaaat ttgctggctt aggcggtggt atggatgaat
     5161 tgtataaggg atccgggtca ggctccggat ctggccctcc cccagctctt ccgccaaaaa
     5221 gacgaaggta atggcagtgc ttttaactaa gaattattag tcttttctgc ttattttttc
     5281 atcatagttt agaacacttt atattaacga atagtttatg aatctattta ggtttaaaaa
     5341 ttgatacagt tttataagtt actttttcaa agactcgtgc tgtctattgc ataatgcact
     5401 ggaaggggaa aaaaaaggtg cacacgcgtg gctttttctt gaatttgcag tttgaaaaat
     5461 gctgagcagt tacagagatg ttacgaacca ctagtgcact gcagtacacg gtttccttga
     5521 aatttttttg attcggtaat ctccgaacag aaggaagaac gaaggaagga gcacagactt
     5581 agattggtat atatacgcat atgtagtgtt gaagaaacat gaaattgccc agtattctta
     5641 acccaactgc acagaacaaa aacgtgcagg aaacgaagat aaatcatgtc gaaagctaca
     5701 tataaggaac gtgctgctac tcatcctagt cctgttgctg ccaagctatt taatatcatg
     5761 cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc gtaccaccaa ggaattactg
     5821 gagttagttg aagcattagg tcccaaaatt tgtttactaa aaacacatgt ggatatcttg
     5881 actgattttt ccatggaggg cacagttaag ccgctaaagg cattatccgc caagtacaat
     5941 tttttactct tcgaagacag aaaatttgct gacattggta atacagtcaa attgcagtac
     6001 tctgcgggtg tatacagaat agcagaatgg gcagacatta cgaatgcaca cggtgtggtg
     6061 ggcccaggta ttgttagcgg tttgaagcag gcggcagaag aagtaacaaa ggaacctaga
     6121 ggccttttga tgttagcaga attgtcatgc aagggctccc tatctactgg agaatatact
     6181 aagggtactg ttgacattgc gaagagcgac aaagattttg ttatcggctt tattgctcaa
     6241 agagacatgg gtggaagaga tgaaggttac gattggttga ttatgacacc cggtgtgggt
     6301 ttagatgaca agggagatgc attgggtcaa cagtatagaa ccgtggatga tgtggtttct
     6361 acaggatctg acattattat tgttggaaga ggactatttg caaagggaag ggatgctaag
     6421 gtagagggtg aacgttacag aaaagcaggc tgggaagcat atttgagaag atgcggccag
     6481 caaaactaaa aaactgtatt ataagtaaat gcatgtatac taaactcaca aattagagct
     6541 tcaatttaat tatatcagtt attacccgag tagagcactt gaatccactg ccccgggaat
     6601 ctcggtcgta atgatttcta taatgacgaa aaaaaaaaaa ttggaaagaa aaagcttcat
     6661 ggcctttata aaaaggaact atccaatacc tcgccagaac caagtaacag tattttacgg
     6721 ggcacaaatc aagaacaata agacaggact gtaaagatgg acgcattgaa ctccaaagaa
     6781 caacaagagt tccaaaaagt agtggaacaa aagcaaatga aggatttcat gcgtttgtac
     6841 tctaatctgg tagaaagatg tttcacagac tgtgtcaatg acttcacaac atcaaagcta
     6901 accaataagg aacaaacatg catcatgaag tgctcagaaa agttcttgaa gcatagcgaa
     6961 cgtgtagggc agcgtttcca agaacaaaac gctgccttgg gacaaggctt gggccgataa
     7021 ggtgtactgg cgtatatata tctaattatg tatctctggt gtagcccatt tttagcatgt
     7081 aaatataaag accgagcggc cgcgtgttac aaccaattaa ccaattctga ttagaaaaac
     7141 tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt
     7201 tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca
     7261 agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc
     7321 ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt
     7381 gagaatggca aaagcttatg catttctttc cagacttgtt caacaggcca gccattacgc
     7441 tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg
     7501 aggcgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg
     7561 cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat
     7621 acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta
     7681 cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc
     7741 atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc
     7801 gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga
     7861 gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg cctggagcaa
     7921 gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac
     7981 agttttattg ttcatgatga tatattttta tcttgtgcaa tgtaacatca gagattttga
     8041 gacacaacgt ggctttgttg aataaatcga acttttgctg agttgaagga tcagtcatga
     8101 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     8161 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     8221 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     8281 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     8341 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     8401 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     8461 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     8521 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     8581 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     8641 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     8701 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     8761 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     8821 tctttcctgc gttatcccct gattctgtgg ataaccgtgc ggccgccaat gctaaattcg
     8881 agtgaaacac aggaagatca gaaaatcctc atttcatcca tattaacaat aatttcaaat
     8941 gtttatttgc attatttgaa actaggcaag acaagcaacg aaacgttttt gaaaattttg
     9001 agtattttca ataaatttgt agaggactca gatattgaaa aaaagctaca gcaattaata
     9061 cttgataaga agagtattga gaagggcaac ggttcatcat ctcatggatc tgcacatgaa
     9121 caaacaccag agtcaaacga cgttgaaatt gaggctactg cgccaattga tgacaataca
     9181 gacgatgata acaaaccgaa gttatctgat gtagaaaagg attaaagatg ctaagagata
     9241 gtgatgatat ttcataaata atgtaattct atatatgtta attacctttt ttgcgaggca
     9301 tatttatggt gaaggataag ttttgaccat caaagaaggt taatgtggct gtggtttcag
     9361 ggtccataaa gcccacatgg ataacattac ccctgaattc gcatctagac tga
//
