LOCUS       sequence-326724-        6353 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  yEVenus under the control of CUP1 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6353)
  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 6353)
  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: pVG45
FEATURES             Location/Qualifiers
     source          1..6353
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..235
                     /gene="S. cerevisiae CUP1"
                     /label=CUP1 promoter
                     /note="inducible metallothionein promoter, regulated by 
                     Cu2+"
     CDS             296..1009
                     /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      1567..1754
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1622..1646)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(1822..1838)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1823..1839)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(1863..1882)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1864..1882)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1892..1909)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1892..1908)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1901..1923)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2049..2504
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2136..2155)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2346..2367
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2635..3438)
                     /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(3439..3654)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(3678..3697)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3797..3819
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3857..3875)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3943..4047
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4048..4116,4117..4908)
                     /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(4266..4285)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5079..5667
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5568..5587
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5821..5838
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5955..5976
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5991..5996,5997..6014,6015..6021)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6029..6045
                     /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     6034..6056
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6053..6069
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6053..6069
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6088..6108
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6090..6108
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ctagttagaa aaagacattt ttgctgtcag tcactgtcaa gagattcttt tgctggcatt
       61 tcttctagaa gcaaaaagag cgatgcgtct tttccgctga accgttccag caaaaaagac
      121 taccaacgca atatggattg tcagaatcat ataaaagaga agcaaataac tccttgtctt
      181 gtatcaattg cattataata tcttcttgtt agtgcaatat catatagaag tcatcgaaat
      241 agatattaag aaaaacaaac tgtacaatca atcaatcaat catcacttaa ttaacatgtc
      301 taaaggtgaa gaattattca ctggtgttgt cccaattttg gttgaattag atggtgatgt
      361 taatggtcac aaattttctg tctccggtga aggtgaaggt gatgctactt acggtaaatt
      421 gaccttaaaa ttgatttgta ctactggtaa attgccagtt ccatggccaa ccttagtcac
      481 tactttaggt tatggtttgc aatgttttgc tagataccca gatcatatga aacaacatga
      541 ctttttcaag tctgccatgc cagaaggtta tgttcaagaa agaactattt ttttcaaaga
      601 tgacggtaac tacaagacca gagctgaagt caagtttgaa ggtgatacct tagttaatag
      661 aatcgaatta aaaggtattg attttaaaga agatggtaac attttaggtc acaaattgga
      721 atacaactat aactctcaca atgtttacat cactgctgac aaacaaaaga atggtatcaa
      781 agctaacttc aaaattagac acaacattga agatggtggt gttcaattag ctgaccatta
      841 tcaacaaaat actccaattg gtgatggtcc agtcttgtta ccagacaacc attacttatc
      901 ctatcaatct gccttatcca aagatccaaa cgaaaagaga gaccacatgg tcttgttaga
      961 atttgttact gctgctggta ttacccatgg tattgatgaa ttgtacaaag catccaactt
     1021 gaacatttcg agaaagctta ccatatcaac cccatcatgc tctttcgaaa attcaaatag
     1081 cacatccatt ccttcgcccg cttcctcatc tcaaagccac actccaatga gaaacatgag
     1141 ctcactctct gataacagcg ttttcagccg gaatatggaa caatcatcac caatcactcc
     1201 aagtatgtac caatttggtc agcagcagtc aaacagtata tgtggtagca ccgttagtgt
     1261 gaatagtctg gtgaatacaa ataacaaaca aaggatctac gaacaaatca cgggtcctaa
     1321 cagcaataac gcaaccaatg attatattga tttgctaaac ctaaatgagt ctaacaagga
     1381 aaaccaaaat cccgcaacgg cgcattacct caatgggggc ccacccaaga caagcttcat
     1441 taaccatgga atgttcccct cgccaactgg gaccataaat agcggtaaat ctagcagtgc
     1501 ctcatcttta atttcttttg gtatgggcaa tacccaagta atatagggcg cgccacttct
     1561 aaataagcga atttcttatg atttatgatt tttattatta aataagttat aaaaaaaata
     1621 agtgtataca aattttaaag tgactcttag gttttaaaac gaaaattctt attcttgagt
     1681 aactctttcc tgtaggtcag gttgctttct caggtatagt atgaggtcgc tcttattgac
     1741 cacacctcta ccggcagatc cgctagggat aacagggtaa tatagatccc ccgggctgca
     1801 ggaattcgat atcaagctta tcgataccgt cgacctcgag ggggggcccg gtacccaatt
     1861 cgccctatag tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact
     1921 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     1981 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     2041 gcgaatggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     2101 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     2161 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     2221 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     2281 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     2341 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     2401 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     2461 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttcctgat gcggtatttt
     2521 ctccttacgc atctgtgcgg tatttcacac cgcatagggt aataactgat ataattaaat
     2581 tgaagctcta atttgtgagt ttagtataca tgcatttact tataatacag ttttttagtt
     2641 ttgctggccg catcttctca aatatgcttc ccagcctgct tttctgtaac gttcaccctc
     2701 taccttagca tcccttccct ttgcaaatag tcctcttcca acaataataa tgtcagatcc
     2761 tgtagagacc acatcatcca cggttctata ctgttgaccc aatgcgtctc ccttgtcatc
     2821 taaacccaca ccgggtgtca taatcaacca atcgtaacct tcatctcttc cacccatgtc
     2881 tctttgagca ataaagccga taacaaaatc tttgtcgctc ttcgcaatgt caacagtacc
     2941 cttagtatat tctccagtag atagggagcc cttgcatgac aattctgcta acatcaaaag
     3001 gcctctaggt tcctttgtta cttcttctgc cgcctgcttc aaaccgctaa caatacctgg
     3061 gcccaccaca ccgtgtgcat tcgtaatgtc tgcccattct gctattctgt atacacccgc
     3121 agagtactgc aatttgactg tattaccaat gtcagcaaat tttctgtctt cgaagagtaa
     3181 aaaattgtac ttggcggata atgcctttag cggcttaact gtgccctcca tggaaaaatc
     3241 agtcaagata tccacatgtg tttttagtaa acaaattttg ggacctaatg cttcaactaa
     3301 ctccagtaat tccttggtgg tacgaacatc caatgaagca cacaagtttg tttgcttttc
     3361 gtgcatgata ttaaatagct tggcagcaac aggactagga tgagtagcag cacgttcctt
     3421 atatgtagct ttcgacatga tttatcttcg tttcctgcag gtttttgttc tgtgcagttg
     3481 ggttaagaat actgggcaat ttcatgtttc ttcaacacta catatgcgta tatataccaa
     3541 tctaagtctg tgctccttcc ttcgttcttc cttctgttcg gagattaccg aatcaaaaaa
     3601 atttcaagga aaccgaaatc aaaaaaaaga ataaaaaaaa aatgatgaat tgaaaaggtg
     3661 gtatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     3721 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     3781 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     3841 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     3901 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     3961 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     4021 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     4081 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     4141 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     4201 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     4261 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     4321 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     4381 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     4441 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     4501 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     4561 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     4621 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     4681 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     4741 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     4801 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     4861 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     4921 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     4981 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     5041 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     5101 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     5161 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     5221 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     5281 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     5341 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     5401 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     5461 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     5521 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     5581 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     5641 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     5701 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     5761 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     5821 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     5881 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     5941 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
     6001 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     6061 agctatgacc atgattacgc caagcgcgca attaaccctc actaaaggga acaaaagctg
     6121 gagctccacc gcggtggcgg ccgccagctg aagcttcgta cgctgcaggt cgacggatcc
     6181 taagccgatc ccattaccga catttgggcg ctatacgtgc atatgttcat gtatgtatct
     6241 gtatttaaaa cacttttgta ttatttttcc tcatatatgt gtataggttt atacggatga
     6301 tttaattatt acttcaccac cctttatttc aggctgatat cttagccttg tta
//
