LOCUS       sequence-475885-        6941 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express Venus as ER marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6941)
  TITLE     Dueber Lab plasmids 2025
REFERENCE   2  (bases 1 to 6941)
  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: pVL1727
FEATURES             Location/Qualifiers
     source          1..6941
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        299..712
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             804..1514
                     /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"
     promoter        2056..2463
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             2464..3558
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     CDS             complement(4589..5404)
                     /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(4589..5404)
                     /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     5312..5331
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5396..5415
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5629..6217
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6118..6137
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cacacactgg cttaagatga caacgccttg ccaacaggga gttcttcaga gacatggagg
       61 ctcaaaacga aattattgac agcctagaca tcaatagtca tacaacagaa agcgaccacc
      121 caactttggc tgataatagc gtataaacaa tgcatacttt gtacgttcaa aatacaatgc
      181 agtagatata tttatgcata ttacatataa tacatatcac ataggaagca acaggcgcgt
      241 tggactttta attttcgagg accgcgaatc cttacatcac acccaatccc ccacaagtga
      301 tcccccacac accatagctt caaaatgttt ctactccttt tttactcttc cagattttct
      361 cggactccgc gcatcgccgt accacttcaa aacacccaag cacagcatac taaatttccc
      421 ctctttcttc ctctagggtg tcgttaatta cccgtactaa aggtttggaa aagaaaaaag
      481 acaccgcctc gtttcttttt cttcgtcgaa aaaggcaata aaaattttta tcacgtttct
      541 ttttcttgaa aatttttttt tttgattttt ttctctttcg atgacctccc attgatattt
      601 aagttaataa acggtcatca atttctcaag tttcagtttc atttttcttg ttctattaca
      661 acttttttta cttcttgctc attagaaaga aagcatagca atctaatcta agttttaatt
      721 acaaaagatc tatgaggcag gtttggttct cttggattgt gggattgttc ctatgttttt
      781 tcaacgtgtc ttctgctggt tcttctaaag gtgaagaatt attcactggt gttgtcccaa
      841 ttttggttga attagatggt gatgttaatg gtcacaaatt ttctgtctcc ggtgaaggtg
      901 aaggtgatgc tacttacggt aaattgacct taaaattgat ttgtactact ggtaaattgc
      961 cagttccatg gccaacctta gtcactactt taggttatgg tttgcaatgt tttgctagat
     1021 acccagatca tatgaaacaa catgactttt tcaagtctgc catgccagaa ggttatgttc
     1081 aagaaagaac tatttttttc aaagatgacg gtaactacaa gaccagagct gaagtcaagt
     1141 ttgaaggtga taccttagtt aatagaatcg aattaaaagg tattgatttt aaagaaggtg
     1201 gtaacatttt aggtcacaaa ttggaataca actataactc tcacaatgtt tacatcactg
     1261 ctgacaaaca aaagaatggt atcaaagcta acttcaaaat tagacacaac attgaagatg
     1321 gtggtgttca attagctgac cattatcaac aaaatactcc aattggtgat ggtccagtct
     1381 tgttaccaga caaccattac ttatcctatc aatctgcctt atccaaagat ccaaacgaaa
     1441 agagagatca catggtcttg ttagaatttg ttactgctgc tggtattacc catggtatgg
     1501 atgaattgta caaaggatcc cacgacgaat tgtagtaatg gcagtgcttt taactaagaa
     1561 ttattagtct tttctgctta ttttttcatc atagtttaga acactttata ttaacgaata
     1621 gtttatgaat ctatttaggt ttaaaaattg atacagtttt ataagttact ttttcaaaga
     1681 ctcgtgctgt ctattgcata atgcactgga aggggaaaaa aaaggtgcac acgcgtggct
     1741 ttttcttgaa tttgcagttt gaaaaatgct gagcagttac agagatgtta cgaaccacta
     1801 gtgcactgca gtacatcgag gagaacttct agtatatcta catacctaat attattgcct
     1861 tattaaaaat ggaatcccaa caattacatc aaaatccaca ttctcttcaa aatcaattgt
     1921 cctgtacttc cttgttcatg tgtgttcaaa aacgttatat ttataggata attatactct
     1981 atttctcaac aagtaattgg ttgtttggcc gagcggtcta aggcgcctga ttcaagaaat
     2041 atcttgaccg cagttaactg tgggaatact caggtatcgt aagatgcaag agttcgaatc
     2101 tcttagcaac cattattttt ttcctcaaca taacgagaac acacaggggc gctatcgcac
     2161 agaatcaaat tcgatgactg gaaatttttt gttaatttca gaggtcgcct gacgcatata
     2221 cctttttcaa ctgaaaaatt gggagaaaaa ggaaaggtga gagcgccgga accggctttt
     2281 catatagaat agagaagcgt tcatgactaa atgcttgcat cacaatactt gaagttgaca
     2341 atattattta aggacctatt gttttttcca ataggtggtt agcaatcgtc ttactttcta
     2401 acttttctta ccttttacat ttcagcaata tatatatata tatttcaagg atataccatt
     2461 ctaatgtctg cccctaagaa gatcgtcgtt ttgccaggtg accacgttgg tcaagaaatc
     2521 acagccgaag ccattaaggt tcttaaagct atttctgatg ttcgttccaa tgtcaagttc
     2581 gatttcgaaa atcatttaat tggtggtgct gctatcgatg ctacaggtgt tccacttcca
     2641 gatgaggcgc tggaagcctc caagaaggct gatgccgttt tgttaggtgc tgtgggtggt
     2701 cctaaatggg gaaccggtag tgttagacct gaacaaggtt tactaaaaat ccgtaaagaa
     2761 cttcaattgt acgccaactt aagaccatgt aactttgcat ccgactctct tttagactta
     2821 tctccaatca agccacaatt tgctaaaggt actgacttcg ttgttgtcag agaattagtg
     2881 ggaggtattt actttggtaa gagaaaggaa gacgatggtg atggtgtcgc ttgggatagt
     2941 gaacaataca ccgttccaga agtgcaaaga atcacaagaa tggccgcttt catggcccta
     3001 caacatgagc caccattgcc tatttggtcc ttggataaag ctaatgtttt ggcctcttca
     3061 agattatgga gaaaaactgt ggaggaaacc atcaagaacg aatttcctac attgaaggtt
     3121 caacatcaat tgattgattc tgccgccatg atcctagtta agaacccaac ccacctaaat
     3181 ggtattataa tcaccagcaa catgtttggt gatatcatct ccgatgaagc ctccgttatc
     3241 ccaggttcct tgggtttgtt gccatctgcg tccttggcct ctttgccaga caagaacacc
     3301 gcatttggtt tgtacgaacc atgccacggt tctgctccag atttgccaaa gaataaggtc
     3361 aaccctatcg ccactatctt gtctgctgca atgatgttga aattgtcatt gaacttgcct
     3421 gaagaaggta aggccattga agatgcagtt aaaaaggttt tggatgcagg tatcagaact
     3481 ggtgatttag gtggttccaa cagtaccacc gaagtcggtg atgctgtcgc cgaagaagtt
     3541 aagaaaatcc ttgcttaaaa agattctctt tttttatgat atttgtacat aaactttata
     3601 aatgaaattc ataatagaaa cgacacgaaa ttacaaaatg gaatatgttc atagggtaga
     3661 cgaaactata tacgcaatct acatacattt atcaagaagg agaaaaagga ggatgtaaag
     3721 gaatacaggt aagcaaattg atactaatgg ctcaacgtga taaggaaaaa gaattgcact
     3781 ttaacattaa tattgacaag gaggagggca ccacacaaaa agttaggtgt aacagaaaat
     3841 catgaaacta tgattcctaa tttatatatt ggaggatttt ctctaaaaaa aaaaaaatac
     3901 aacaaataaa aaacactcaa tgacctgacc atttgatgga gtttaagtca ataccttctt
     3961 gaaccatttc ccataatggt gaaagttccc tcaagaattt tactctgtca gaaacggcct
     4021 taacgacgta gtcgagagta ctcgtatcgc atgtcggtgc gacacgaaat tacaaaatgg
     4081 aatatgttca tagggtagac gaaactatat acgcaatcta catacattta tcaagaagga
     4141 gaaaaaggag gatgtaaagg aatacaggta agcaaattga tactaatggc tcaacgtgat
     4201 aaggaaaaag aattgcactt taacattaat attgacaagg aggagggcac cacacaaaaa
     4261 gttaggtgta acagaaaatc atgaaactat gattcctaat ttatatattg gaggattttc
     4321 tctaaaaaaa aaaaaataca acaaataaaa aacactcaat gacctgacca tttgatggag
     4381 tttaagtcaa taccttcttg aaccatttcc cataatggtg aaagttccct caagaatttt
     4441 actctgtcag aaacggcctt aacgacgtag tcgacctcct cttcagtact aaatctacca
     4501 ataccaaatc tgatggaaga atgggctaat gcatcatcct tacccagcgc cgagcggccg
     4561 cgtgttacaa ccaattaacc aattctgatt agaaaaactc atcgagcatc aaatgaaact
     4621 gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg
     4681 aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga
     4741 ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat
     4801 caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa agcttatgca
     4861 tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat
     4921 caaccaaacc gttattcatt cgtgattgcg cctgagcgag gcgaaatacg cgatcgctgt
     4981 taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat
     5041 caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg
     5101 ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg
     5161 gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg
     5221 caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc
     5281 gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat
     5341 cagcatccat gttggaattt aatcgcggcc tggagcaaga cgtttcccgt tgaatatggc
     5401 tcataacacc ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgatgata
     5461 tatttttatc ttgtgcaatg taacatcaga gattttgaga cacaacgtgg ctttgttgaa
     5521 taaatcgaac ttttgctgag ttgaaggatc agtcatgacc aaaatccctt aacgtgagtt
     5581 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5641 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5701 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     5761 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5821 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     5881 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5941 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     6001 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6061 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6121 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6181 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     6241 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     6301 ttctgtggat aaccgtgcgg ccgccaatac ccagaagaac agtaaaataa agcaaggtac
     6361 gtgaaattaa tatttttaaa tggttctaac cgatgccgaa gaactgcgca gtccggttat
     6421 aacgtctgac atgtcctttt ttgatttgga atccaaccac tcaagtgact ctgttcattt
     6481 actttgcgaa aaatataccc acaaattgcc catcgaaagt gaatcgcaaa ccaccttcag
     6541 actggcaccg acaaagcaaa gattatacag acagagtact ttatacgtac cgttaagtct
     6601 caagcaaagg gttttcttat ttactgaacg ggtaaagagt atctgggccg gcttgccaag
     6661 atgcaaaccg aataagtatt tcaaagttgc atttgcctta gccgtcctga caccattggc
     6721 tatttggata ttttatattg actttcgtgt acattgatca catcgactgt tctattggca
     6781 aatgaaccac gggcattgac tatttttcag gttactacta tatattatca tcacgggcaa
     6841 ggattgtacc ctgaattcgc atctagactg aactggccga taattgcaga cgaacgacta
     6901 gagatctatg tgaggatcct aactcgagat cgagctggat g
//
