LOCUS       sequence-326984-        6349 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  yEVenus under the control of tetOpr with ADH1t upstream of the 
            reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6349)
  AUTHORS   Gligorovski V, Sadeghi A, Rahi SJ
  TITLE     Multidimensional characterization of inducible promoters and a 
            highly light-sensitive LOV-transcription factor.
  JOURNAL   Nat Commun. 2023 Jun 27;14(1):3810. doi: 10.1038/s41467-023-38959-8.
  PUBMED    37369667
REFERENCE   2  (bases 1 to 6349)
  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"; journalName: "Nat 
            Commun."; date: "2023-06-27"; volume: "14(1):3810. doi"; pages: " 
            10.1038/s41467-023-38959-8"
COMMENT     Sequence Label: pVG50
FEATURES             Location/Qualifiers
     source          1..6349
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    123..144
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(159..164,165..182,183..189)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    197..213
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     202..224
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     221..237
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     221..237
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     256..276
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        258..276
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     terminator      365..552
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     protein_bind    597..615
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             813..1526
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGIDELYK"
     terminator      2084..2271
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2339..2355)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2340..2356)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(2380..2399)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2381..2399)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2409..2426)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2409..2425)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2418..2440)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2566..3021
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2653..2672)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2863..2884
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3152..3955)
                     /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"
     promoter        complement(3956..4171)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4195..4214)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4314..4336
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4374..4392)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4460..4564
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4565..4633,4634..5425)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4783..4802)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5596..6184
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6085..6104
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     join(6338..6349,1..6)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
       61 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
      121 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
      181 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
      241 gattacgcca agcgcgcaat taaccctcac taaagggaac aaaagctgga gctccaccgc
      301 ggtggcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccg ccacttctaa
      361 ataagcgaat ttcttatgat ttatgatttt tattattaaa taagttataa aaaaaataag
      421 tgtatacaaa ttttaaagtg actcttaggt tttaaaacga aaattcttat tcttgagtaa
      481 ctctttcctg taggtcaggt tgctttctca ggtatagtat gaggtcgctc ttattgacca
      541 cacctctacc ggcagatccg ctagggataa cagggtaata tagatcaatt cctcgatccc
      601 tatcagtgat agagagtcga caaagtcgag tttctcgatc gagaccactg catgcatgtg
      661 ctctgtatgt atataaaact cttgttttct tcttttctct aaatattctt tccttataca
      721 ttaggtcctt tgtagcataa attactatac ttctatagac acgcaaacac aaatacacac
      781 actaaattac cggatcaatt cggttaatta acatgtctaa aggtgaagaa ttattcactg
      841 gtgttgtccc aattttggtt gaattagatg gtgatgttaa tggtcacaaa ttttctgtct
      901 ccggtgaagg tgaaggtgat gctacttacg gtaaattgac cttaaaattg atttgtacta
      961 ctggtaaatt gccagttcca tggccaacct tagtcactac tttaggttat ggtttgcaat
     1021 gttttgctag atacccagat catatgaaac aacatgactt tttcaagtct gccatgccag
     1081 aaggttatgt tcaagaaaga actatttttt tcaaagatga cggtaactac aagaccagag
     1141 ctgaagtcaa gtttgaaggt gataccttag ttaatagaat cgaattaaaa ggtattgatt
     1201 ttaaagaaga tggtaacatt ttaggtcaca aattggaata caactataac tctcacaatg
     1261 tttacatcac tgctgacaaa caaaagaatg gtatcaaagc taacttcaaa attagacaca
     1321 acattgaaga tggtggtgtt caattagctg accattatca acaaaatact ccaattggtg
     1381 atggtccagt cttgttacca gacaaccatt acttatccta tcaatctgcc ttatccaaag
     1441 atccaaacga aaagagagac cacatggtct tgttagaatt tgttactgct gctggtatta
     1501 cccatggtat tgatgaattg tacaaagcat ccaacttgaa catttcgaga aagcttacca
     1561 tatcaacccc atcatgctct ttcgaaaatt caaatagcac atccattcct tcgcccgctt
     1621 cctcatctca aagccacact ccaatgagaa acatgagctc actctctgat aacagcgttt
     1681 tcagccggaa tatggaacaa tcatcaccaa tcactccaag tatgtaccaa tttggtcagc
     1741 agcagtcaaa cagtatatgt ggtagcaccg ttagtgtgaa tagtctggtg aatacaaata
     1801 acaaacaaag gatctacgaa caaatcacgg gtcctaacag caataacgca accaatgatt
     1861 atattgattt gctaaaccta aatgagtcta acaaggaaaa ccaaaatccc gcaacggcgc
     1921 attacctcaa tgggggccca cccaagacaa gcttcattaa ccatggaatg ttcccctcgc
     1981 caactgggac cataaatagc ggtaaatcta gcagtgcctc atctttaatt tcttttggta
     2041 tgggcaatac ccaagtaata tagggcgcgc cacttctaaa taagcgaatt tcttatgatt
     2101 tatgattttt attattaaat aagttataaa aaaaataagt gtatacaaat tttaaagtga
     2161 ctcttaggtt ttaaaacgaa aattcttatt cttgagtaac tctttcctgt aggtcaggtt
     2221 gctttctcag gtatagtatg aggtcgctct tattgaccac acctctaccg gcagatccgc
     2281 tagggataac agggtaatat agatcccccg ggctgcagga attcgatatc aagcttatcg
     2341 ataccgtcga cctcgagggg gggcccggta cccaattcgc cctatagtga gtcgtattac
     2401 gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     2461 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     2521 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggacgcg ccctgtagcg
     2581 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     2641 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     2701 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     2761 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     2821 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     2881 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     2941 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     3001 aaatattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     3061 ttcacaccgc atagggtaat aactgatata attaaattga agctctaatt tgtgagttta
     3121 gtatacatgc atttacttat aatacagttt tttagttttg ctggccgcat cttctcaaat
     3181 atgcttccca gcctgctttt ctgtaacgtt caccctctac cttagcatcc cttccctttg
     3241 caaatagtcc tcttccaaca ataataatgt cagatcctgt agagaccaca tcatccacgg
     3301 ttctatactg ttgacccaat gcgtctccct tgtcatctaa acccacaccg ggtgtcataa
     3361 tcaaccaatc gtaaccttca tctcttccac ccatgtctct ttgagcaata aagccgataa
     3421 caaaatcttt gtcgctcttc gcaatgtcaa cagtaccctt agtatattct ccagtagata
     3481 gggagccctt gcatgacaat tctgctaaca tcaaaaggcc tctaggttcc tttgttactt
     3541 cttctgccgc ctgcttcaaa ccgctaacaa tacctgggcc caccacaccg tgtgcattcg
     3601 taatgtctgc ccattctgct attctgtata cacccgcaga gtactgcaat ttgactgtat
     3661 taccaatgtc agcaaatttt ctgtcttcga agagtaaaaa attgtacttg gcggataatg
     3721 cctttagcgg cttaactgtg ccctccatgg aaaaatcagt caagatatcc acatgtgttt
     3781 ttagtaaaca aattttggga cctaatgctt caactaactc cagtaattcc ttggtggtac
     3841 gaacatccaa tgaagcacac aagtttgttt gcttttcgtg catgatatta aatagcttgg
     3901 cagcaacagg actaggatga gtagcagcac gttccttata tgtagctttc gacatgattt
     3961 atcttcgttt cctgcaggtt tttgttctgt gcagttgggt taagaatact gggcaatttc
     4021 atgtttcttc aacactacat atgcgtatat ataccaatct aagtctgtgc tccttccttc
     4081 gttcttcctt ctgttcggag attaccgaat caaaaaaatt tcaaggaaac cgaaatcaaa
     4141 aaaaagaata aaaaaaaaat gatgaattga aaaggtggta tggtgcactc tcagtacaat
     4201 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     4261 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     4321 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     4381 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     4441 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     4501 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     4561 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     4621 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     4681 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     4741 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     4801 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     4861 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     4921 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     4981 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     5041 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     5101 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     5161 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     5221 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     5281 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     5341 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     5401 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     5461 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     5521 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     5581 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5641 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5701 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     5761 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5821 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     5881 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     5941 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6001 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6061 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6121 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6181 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6241 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     6301 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtg
//
