LOCUS       sequence-476352-        8563 bp    DNA     circular SYN 18-JUN-2025
DEFINITION  Express CpPT1 membrane protein cargo with C-terminal mTurqoise2 
            marker but without any targeting sequence and Venus ER marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8563)
  TITLE     Dueber Lab plasmids 2025
REFERENCE   2  (bases 1 to 8563)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 18, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pVL1172
FEATURES             Location/Qualifiers
     source          1..8563
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3072..3485
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             3577..4287
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKL
                     ICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEGGNILGHKLEYNYNSHNVYITADKQKNGIKAN
                     FKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     CDS             4766..5569
                     /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(6211..7026)
                     /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(6211..7026)
                     /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     6934..6953
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     7018..7037
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      7251..7839
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7740..7759
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 accagatgtc aacacagcta caacgttgat 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 tatgattcag tcttcattct gtttaacaaa taataagata ggtaacaatg
      781 aagatatgat gaatagatac cataaaccat taaagaaatc tacagttcct atggctttac
      841 aagatgactc tgctacaaaa tctcaaaacg aaaatattgt ttctacatct ttcttggact
      901 tcttaaccaa gaagttggat gcattttata ggttatcaag accatatgct tggacttcaa
      961 tcattgtagg tattttgtca tcttctttgt tgccaatcca atctttggca gatttgactc
     1021 ctaccttctt gatcgaggta ttgaagccta tagttccttc aattatgatg aatatatttg
     1081 tcgttgctat taatcagtta tcagatatag caattgataa ggttaacaag ccaaacttgc
     1141 ctttagtttc tggtgacata tctattggtg aggctatagc tattgctgtt atctcaacat
     1201 tgacttcttt ggctatgggt gttatgttaa gatcaccacc attagttata gcattaatat
     1261 tgaggtgtat cttaggtgca gcttattcta ttgatttgcc attattgaga tggaaggctt
     1321 ctcctttgat ggctgccgtc gcaatcgtta ttggaaatgg tataaataat gttttgccat
     1381 acttcttaca cgtccaaaag tacgttttag gtaggcctgt tgtctttact aagcctttat
     1441 tattcgcagt tgcattctca gcaatatttt ctatcgtctt atcattctta aaagacattc
     1501 ctgacgttga aggagataag aaatctggta tcagaacatt accagtcatt ttgggtaaag
     1561 agagagtctt gtctatgtca actggtattt tgttgatggc ctatgcttct gccgccttgg
     1621 ctggagtctt ctcaccaatt ttattatgta aattggtaac tatgattggt cattcagtat
     1681 tgggttttat cttatggtct aaggcccaaa cagttgattt atcaaatgct aaatctacat
     1741 attcattctt cttattcatt ttccagttgt attacaccga gttcttctta atgcattttg
     1801 taagaggttc tgtttctaaa ggtgaagaat tattcactgg tgttgtccca attttggttg
     1861 aattagatgg tgatgttaat ggtcacaaat tttctgtctc cggtgaaggt gaaggtgatg
     1921 ctacttacgg taaattgacc ttaaaattta tttgtactac tggtaaattg ccagttccat
     1981 ggccaacctt agtcactact ttatcttggg gtgttcaatg ttttgcaaga tacccagatc
     2041 atatgaaaca acatgacttt ttcaagtctg ccatgccaga aggttatgtt caagaaagaa
     2101 ctattttttt caaagatgac ggtaactaca agaccagagc tgaagtcaag tttgaaggtg
     2161 ataccttagt taatagaatc gaattaaaag gtattgattt taaagaagat ggtaacattt
     2221 taggtcacaa attggaatac aattatttct ctgacaatgt ttacatcact gctgacaaac
     2281 aaaagaatgg tatcaaagct aacttcaaaa ttagacacaa cattgaagat ggtggtgttc
     2341 aattagctga ccattatcaa caaaatactc caattggtga tggtccagtc ttgttaccag
     2401 acaaccatta cttatccact caatctaagt tatccaaaga tccaaacgaa aagagggacc
     2461 acatggtctt gttagaattt gttactgctg ctggtattac cttgggtatg gatgaattgt
     2521 acaaaggatc ctaactcgag gccaattggt gcggcaattg ataataacga aaatgtcttt
     2581 taatgatctg ggtataatga ggaattttcc gaacgttttt actttatata tatatataca
     2641 tgtaacatat attctatacg ctatagagaa aggaaatttt tcaattaaaa aaaaaataga
     2701 gaaagagttt cacttcttga ttatcgctaa cactaatggt tgaagtactg ctactttaat
     2761 tttatgctgg atgcacacac tggcttaaga tgacaacgcc ttgccaacag ggagttcttc
     2821 agagacatgg aggctcaaaa cgaaattatt gacagcctag acatcaatag tcatacaaca
     2881 gaaagcgacc acccaacttt ggctgataat agcgtataaa caatgcatac tttgtacgtt
     2941 caaaatacaa tgcagtagat atatttatgc atattacata taatacatat cacataggaa
     3001 gcaacaggcg cgttggactt ttaattttcg aggaccgcga atccttacat cacacccaat
     3061 cccccacaag tgatccccca cacaccatag cttcaaaatg tttctactcc ttttttactc
     3121 ttccagattt tctcggactc cgcgcatcgc cgtaccactt caaaacaccc aagcacagca
     3181 tactaaattt cccctctttc ttcctctagg gtgtcgttaa ttacccgtac taaaggtttg
     3241 gaaaagaaaa aagacaccgc ctcgtttctt tttcttcgtc gaaaaaggca ataaaaattt
     3301 ttatcacgtt tctttttctt gaaaattttt ttttttgatt tttttctctt tcgatgacct
     3361 cccattgata tttaagttaa taaacggtca tcaatttctc aagtttcagt ttcatttttc
     3421 ttgttctatt acaacttttt ttacttcttg ctcattagaa agaaagcata gcaatctaat
     3481 ctaagtttta attacaaaag atctatgagg caggtttggt tctcttggat tgtgggattg
     3541 ttcctatgtt ttttcaacgt gtcttctgct ggttcttcta aaggtgaaga attattcact
     3601 ggtgttgtcc caattttggt tgaattagat ggtgatgtta atggtcacaa attttctgtc
     3661 tccggtgaag gtgaaggtga tgctacttac ggtaaattga ccttaaaatt gatttgtact
     3721 actggtaaat tgccagttcc atggccaacc ttagtcacta ctttaggtta tggtttgcaa
     3781 tgttttgcta gatacccaga tcatatgaaa caacatgact ttttcaagtc tgccatgcca
     3841 gaaggttatg ttcaagaaag aactattttt ttcaaagatg acggtaacta caagaccaga
     3901 gctgaagtca agtttgaagg tgatacctta gttaatagaa tcgaattaaa aggtattgat
     3961 tttaaagaag gtggtaacat tttaggtcac aaattggaat acaactataa ctctcacaat
     4021 gtttacatca ctgctgacaa acaaaagaat ggtatcaaag ctaacttcaa aattagacac
     4081 aacattgaag atggtggtgt tcaattagct gaccattatc aacaaaatac tccaattggt
     4141 gatggtccag tcttgttacc agacaaccat tacttatcct atcaatctgc cttatccaaa
     4201 gatccaaacg aaaagagaga tcacatggtc ttgttagaat ttgttactgc tgctggtatt
     4261 acccatggta tggatgaatt gtacaaagga tcccacgacg aattgtagta atggcagtgc
     4321 ttttaactaa gaattattag tcttttctgc ttattttttc atcatagttt agaacacttt
     4381 atattaacga atagtttatg aatctattta ggtttaaaaa ttgatacagt tttataagtt
     4441 actttttcaa agactcgtgc tgtctattgc ataatgcact ggaaggggaa aaaaaaggtg
     4501 cacacgcgtg gctttttctt gaatttgcag tttgaaaaat gctgagcagt tacagagatg
     4561 ttacgaacca ctagtgcact gcagtacacg gtttccttga aatttttttg attcggtaat
     4621 ctccgaacag aaggaagaac gaaggaagga gcacagactt agattggtat atatacgcat
     4681 atgtagtgtt gaagaaacat gaaattgccc agtattctta acccaactgc acagaacaaa
     4741 aacgtgcagg aaacgaagat aaatcatgtc gaaagctaca tataaggaac gtgctgctac
     4801 tcatcctagt cctgttgctg ccaagctatt taatatcatg cacgaaaagc aaacaaactt
     4861 gtgtgcttca ttggatgttc gtaccaccaa ggaattactg gagttagttg aagcattagg
     4921 tcccaaaatt tgtttactaa aaacacatgt ggatatcttg actgattttt ccatggaggg
     4981 cacagttaag ccgctaaagg cattatccgc caagtacaat tttttactct tcgaagacag
     5041 aaaatttgct gacattggta atacagtcaa attgcagtac tctgcgggtg tatacagaat
     5101 agcagaatgg gcagacatta cgaatgcaca cggtgtggtg ggcccaggta ttgttagcgg
     5161 tttgaagcag gcggcagaag aagtaacaaa ggaacctaga ggccttttga tgttagcaga
     5221 attgtcatgc aagggctccc tatctactgg agaatatact aagggtactg ttgacattgc
     5281 gaagagcgac aaagattttg ttatcggctt tattgctcaa agagacatgg gtggaagaga
     5341 tgaaggttac gattggttga ttatgacacc cggtgtgggt ttagatgaca agggagatgc
     5401 attgggtcaa cagtatagaa ccgtggatga tgtggtttct acaggatctg acattattat
     5461 tgttggaaga ggactatttg caaagggaag ggatgctaag gtagagggtg aacgttacag
     5521 aaaagcaggc tgggaagcat atttgagaag atgcggccag caaaactaaa aaactgtatt
     5581 ataagtaaat gcatgtatac taaactcaca aattagagct tcaatttaat tatatcagtt
     5641 attacccgag tagagcactt gaatccactg ccccgggaat ctcggtcgta atgatttcta
     5701 taatgacgaa aaaaaaaaaa ttggaaagaa aaagcttcat ggcctttata aaaaggaact
     5761 atccaatacc tcgccagaac caagtaacag tattttacgg ggcacaaatc aagaacaata
     5821 agacaggact gtaaagatgg acgcattgaa ctccaaagaa caacaagagt tccaaaaagt
     5881 agtggaacaa aagcaaatga aggatttcat gcgtttgtac tctaatctgg tagaaagatg
     5941 tttcacagac tgtgtcaatg acttcacaac atcaaagcta accaataagg aacaaacatg
     6001 catcatgaag tgctcagaaa agttcttgaa gcatagcgaa cgtgtagggc agcgtttcca
     6061 agaacaaaac gctgccttgg gacaaggctt gggccgataa ggtgtactgg cgtatatata
     6121 tctaattatg tatctctggt gtagcccatt tttagcatgt aaatataaag accgagcggc
     6181 cgcgtgttac aaccaattaa ccaattctga ttagaaaaac tcatcgagca tcaaatgaaa
     6241 ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa
     6301 tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc
     6361 gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt
     6421 atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagcttatg
     6481 catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc
     6541 atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg aggcgaaata cgcgatcgct
     6601 gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc
     6661 atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg ctgttttccc
     6721 ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt
     6781 cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt
     6841 ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa
     6901 tcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa
     6961 atcagcatcc atgttggaat ttaatcgcgg cctggagcaa gacgtttccc gttgaatatg
     7021 gctcataaca ccccttgtat tactgtttat gtaagcagac agttttattg ttcatgatga
     7081 tatattttta tcttgtgcaa tgtaacatca gagattttga gacacaacgt ggctttgttg
     7141 aataaatcga acttttgctg agttgaagga tcagtcatga ccaaaatccc ttaacgtgag
     7201 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     7261 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     7321 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     7381 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     7441 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     7501 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     7561 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     7621 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     7681 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     7741 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     7801 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     7861 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     7921 gattctgtgg ataaccgtgc ggccgccaat gctaaattcg agtgaaacac aggaagatca
     7981 gaaaatcctc atttcatcca tattaacaat aatttcaaat gtttatttgc attatttgaa
     8041 actaggcaag acaagcaacg aaacgttttt gaaaattttg agtattttca ataaatttgt
     8101 agaggactca gatattgaaa aaaagctaca gcaattaata cttgataaga agagtattga
     8161 gaagggcaac ggttcatcat ctcatggatc tgcacatgaa caaacaccag agtcaaacga
     8221 cgttgaaatt gaggctactg cgccaattga tgacaataca gacgatgata acaaaccgaa
     8281 gttatctgat gtagaaaagg attaaagatg ctaagagata gtgatgatat ttcataaata
     8341 atgtaattct atatatgtta attacctttt ttgcgaggca tatttatggt gaaggataag
     8401 ttttgaccat caaagaaggt taatgtggct gtggtttcag ggtccataaa gcccacatgg
     8461 ataacattac ccctgaattc gcatctagac tgaactggcc gataattgca gacgaacgac
     8521 tagagatcta tgtgaggatc ctaactcgag atcgagctgc caa
//
