LOCUS       sequence-475869-        5790 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 5790)
  TITLE     Dueber Lab plasmids 2025
REFERENCE   2  (bases 1 to 5790)
  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: pVL1176
FEATURES             Location/Qualifiers
     source          1..5790
                     /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"
     CDS             1993..2796
                     /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(3438..4253)
                     /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(3438..4253)
                     /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     4161..4180
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4245..4264
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4478..5066
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4967..4986
                     /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 gtacacggtt tccttgaaat ttttttgatt cggtaatctc cgaacagaag
     1861 gaagaacgaa ggaaggagca cagacttaga ttggtatata tacgcatatg tagtgttgaa
     1921 gaaacatgaa attgcccagt attcttaacc caactgcaca gaacaaaaac gtgcaggaaa
     1981 cgaagataaa tcatgtcgaa agctacatat aaggaacgtg ctgctactca tcctagtcct
     2041 gttgctgcca agctatttaa tatcatgcac gaaaagcaaa caaacttgtg tgcttcattg
     2101 gatgttcgta ccaccaagga attactggag ttagttgaag cattaggtcc caaaatttgt
     2161 ttactaaaaa cacatgtgga tatcttgact gatttttcca tggagggcac agttaagccg
     2221 ctaaaggcat tatccgccaa gtacaatttt ttactcttcg aagacagaaa atttgctgac
     2281 attggtaata cagtcaaatt gcagtactct gcgggtgtat acagaatagc agaatgggca
     2341 gacattacga atgcacacgg tgtggtgggc ccaggtattg ttagcggttt gaagcaggcg
     2401 gcagaagaag taacaaagga acctagaggc cttttgatgt tagcagaatt gtcatgcaag
     2461 ggctccctat ctactggaga atatactaag ggtactgttg acattgcgaa gagcgacaaa
     2521 gattttgtta tcggctttat tgctcaaaga gacatgggtg gaagagatga aggttacgat
     2581 tggttgatta tgacacccgg tgtgggttta gatgacaagg gagatgcatt gggtcaacag
     2641 tatagaaccg tggatgatgt ggtttctaca ggatctgaca ttattattgt tggaagagga
     2701 ctatttgcaa agggaaggga tgctaaggta gagggtgaac gttacagaaa agcaggctgg
     2761 gaagcatatt tgagaagatg cggccagcaa aactaaaaaa ctgtattata agtaaatgca
     2821 tgtatactaa actcacaaat tagagcttca atttaattat atcagttatt acccgagtag
     2881 agcacttgaa tccactgccc cgggaatctc ggtcgtaatg atttctataa tgacgaaaaa
     2941 aaaaaaattg gaaagaaaaa gcttcatggc ctttataaaa aggaactatc caatacctcg
     3001 ccagaaccaa gtaacagtat tttacggggc acaaatcaag aacaataaga caggactgta
     3061 aagatggacg cattgaactc caaagaacaa caagagttcc aaaaagtagt ggaacaaaag
     3121 caaatgaagg atttcatgcg tttgtactct aatctggtag aaagatgttt cacagactgt
     3181 gtcaatgact tcacaacatc aaagctaacc aataaggaac aaacatgcat catgaagtgc
     3241 tcagaaaagt tcttgaagca tagcgaacgt gtagggcagc gtttccaaga acaaaacgct
     3301 gccttgggac aaggcttggg ccgataaggt gtactggcgt atatatatct aattatgtat
     3361 ctctggtgta gcccattttt agcatgtaaa tataaagacc gagcggccgc gtgttacaac
     3421 caattaacca attctgatta gaaaaactca tcgagcatca aatgaaactg caatttattc
     3481 atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac
     3541 tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt
     3601 ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa
     3661 tcaccatgag tgacgactga atccggtgag aatggcaaaa gcttatgcat ttctttccag
     3721 acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg
     3781 ttattcattc gtgattgcgc ctgagcgagg cgaaatacgc gatcgctgtt aaaaggacaa
     3841 ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt
     3901 tcacctgaat caggatattc ttctaatacc tggaatgctg ttttcccggg gatcgcagtg
     3961 gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata
     4021 aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct
     4081 ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc
     4141 gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg
     4201 ttggaattta atcgcggcct ggagcaagac gtttcccgtt gaatatggct cataacaccc
     4261 cttgtattac tgtttatgta agcagacagt tttattgttc atgatgatat atttttatct
     4321 tgtgcaatgt aacatcagag attttgagac acaacgtggc tttgttgaat aaatcgaact
     4381 tttgctgagt tgaaggatca gtcatgacca aaatccctta acgtgagttt tcgttccact
     4441 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     4501 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     4561 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     4621 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     4681 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     4741 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     4801 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     4861 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     4921 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     4981 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     5041 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     5101 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     5161 accgtgcggc cgccaatgct aaattcgagt gaaacacagg aagatcagaa aatcctcatt
     5221 tcatccatat taacaataat ttcaaatgtt tatttgcatt atttgaaact aggcaagaca
     5281 agcaacgaaa cgtttttgaa aattttgagt attttcaata aatttgtaga ggactcagat
     5341 attgaaaaaa agctacagca attaatactt gataagaaga gtattgagaa gggcaacggt
     5401 tcatcatctc atggatctgc acatgaacaa acaccagagt caaacgacgt tgaaattgag
     5461 gctactgcgc caattgatga caatacagac gatgataaca aaccgaagtt atctgatgta
     5521 gaaaaggatt aaagatgcta agagatagtg atgatatttc ataaataatg taattctata
     5581 tatgttaatt accttttttg cgaggcatat ttatggtgaa ggataagttt tgaccatcaa
     5641 agaaggttaa tgtggctgtg gtttcagggt ccataaagcc cacatggata acattacccc
     5701 tgaattcgca tctagactga actggccgat aattgcagac gaacgactag agatctatgt
     5761 gaggatccta actcgagatc gagctggatg
//
