LOCUS       sequence-258777-        6495 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reporter plasmid, pZF(4BSc)-yEVENUS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6495)
  AUTHORS   An-Adirekkun JM, Stewart CJ, Geller SH, Patel MT, Melendez J, Oakes 
            BL, Noyes MB, McClean MN
  TITLE     A yeast optogenetic toolkit (yOTK) for gene expression control in 
            Saccharomyces cerevisiae.
  JOURNAL   Biotechnol Bioeng. 2019 Dec 2. doi: 10.1002/bit.27234.
  PUBMED    31788779
REFERENCE   2  (bases 1 to 6495)
  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: "Biotechnol 
            Bioeng. 2019 Dec 2. doi: 10.1002/bit.27234."
COMMENT     Sequence Label: pMM0289
FEATURES             Location/Qualifiers
     source          1..6495
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     27..46
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        27..45
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     716..739
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     758..774
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     759..775
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             808..1524
                     /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      1548..1735
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1603..1627)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(1783..1803)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1783..1801)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1822..1838)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1822..1838)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1835..1857)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1846..1862
                     /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)."
     promoter        complement(join(1870..1876,1877..1894,1895..1900))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1915..1936
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2053..2070)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2224..2812)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2304..2323)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2983..3774,3775..3843))
                     /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     3606..3625
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3844..3948)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    3985..4488
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     4529..4547
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4585..4607)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4707..4726
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4750..4965
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             4966..5769
                     /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"
     rep_origin      complement(5900..6355)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6037..6058)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6249..6268
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     join(6481..6495,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(6495,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgag cgcgcgtaat acgactcact atagggcgaa ttgggtacgt
       61 tgctagcttc gtaccttata ttgaattttc aaaaattctt actttttttt tggatggacg
      121 caaagaagtt taataatcat attacatggc attaccacca tatacatatc catatacata
      181 tccatatcta atcttactta tatgttgtgg aaatgtaaag agccccatta tcttagccta
      241 aaaaaacctt ctctttggaa ctttcagtaa tacgcttaac tgctcattgc tatattgaag
      301 tgcggccgcg cgtgggcgtg cgtgggcgtg cgtgggcgtg cgtgggcgtc tagaccgtgc
      361 gtcctcgtct tcaccggtcg cgttcctgaa acgcagatgt gcctcgcgcc gcactgctcc
      421 gaacaataaa gattctacaa tactagcttt tatggttatg aagaggaaaa attggcagta
      481 acctggcccc acaaaccttc aaattaacga atcaaattaa caaccatagg atgataatgc
      541 gattagtttt ttagccttat ttctggggta attaatcagc gaagcgatga tttttgatct
      601 attaacagat atataaatgg aaaagctgca taaccacttt aactaatact ttcaacattt
      661 tcagtttgta ttacttctta ttcaaatgtc ataaaagtat caacaaaaaa ttgttaatat
      721 acctctatac tttaacgtca aggagaaaaa actatactcg aggtcgacgg tatcgataag
      781 cttgatatcg aattcctgca gcccgggatg tctaaaggtg aagaattatt cactggtgtt
      841 gtcccaattt tggttgaatt agatggtgat gttaatggtc acaaattttc tgtctccggt
      901 gaaggtgaag gtgatgctac ttacggtaaa ttgaccttaa aattgatttg tactactggt
      961 aaattgccag ttccatggcc aaccttagtc actactttag gttatggttt gcaatgtttt
     1021 gctagatacc cagatcatat gaaacaacat gactttttca agtctgccat gccagaaggt
     1081 tatgttcaag aaagaactat ttttttcaaa gatgacggta actacaagac cagagctgaa
     1141 gtcaagtttg aaggtgatac cttagttaat agaatcgaat taaaaggtat tgattttaaa
     1201 gaagatggta acattttagg tcacaaattg gaatacaact ataactctca caatgtttac
     1261 atcactgctg acaaacaaaa gaatggtatc aaagctaact tcaaaattag acacaacatt
     1321 gaagatggtg gtgttcaatt agctgaccat tatcaacaaa atactccaat tggtgatggt
     1381 ccagtcttgt taccagacaa ccattactta tcctatcaat ctgccttatc caaagatcca
     1441 aacgaaaaga gagaccacat ggtcttgtta gaatttgtta ctgctgctgg tattacccat
     1501 ggtattgatg aattgtacaa ataatagggc gcgccacttc taaataagcg aatttcttat
     1561 gatttatgat ttttattatt aaataagtta taaaaaaaat aagtgtatac aaattttaaa
     1621 gtgactctta ggttttaaaa cgaaaattct tattcttgag taactctttc ctgtaggtca
     1681 ggttgctttc tcaggtatag tatgaggtcg ctcttattga ccacacctct accggcagat
     1741 ccgctaggga taacagggta atatgagctc cagcttttgt tccctttagt gagggttaat
     1801 tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     1861 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     1921 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     1981 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     2041 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     2101 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     2161 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     2221 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     2281 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     2341 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2401 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     2461 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2521 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     2581 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     2641 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     2701 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2761 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     2821 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     2881 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     2941 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     3001 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     3061 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     3121 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     3181 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     3241 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     3301 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     3361 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     3421 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     3481 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     3541 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     3601 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     3661 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     3721 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     3781 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     3841 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     3901 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     3961 aaaagtgcca cctgggtcct tttcatcacg tgctataaaa ataattataa tttaaatttt
     4021 ttaatataaa tatataaatt aaaaatagaa agtaaaaaaa gaaattaaag aaaaaatagt
     4081 ttttgttttc cgaagatgta aaagactcta gggggatcgc caacaaatac taccttttat
     4141 cttgctcttc ctgctctcag gtattaatgc cgaattgttt catcttgtct gtgtagaaga
     4201 ccacacacga aaatcctgtg attttacatt ttacttatcg ttaatcgaat gtatatctat
     4261 ttaatctgct tttcttgtct aataaatata tatgtaaagt acgctttttg ttgaaatttt
     4321 ttaaaccttt gtttattttt ttttcttcat tccgtaactc ttctaccttc tttatttact
     4381 ttctaaaatc caaatacaaa acataaaaat aaataaacac agagtaaatt cccaaattat
     4441 tccatcatta aaagatacga ggcgcgtgta agttacaggc aagcgatccg tctaagaaac
     4501 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     4561 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     4621 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     4681 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     4741 taccaccttt tcaattcatc attttttttt tattcttttt tttgatttcg gtttccttga
     4801 aatttttttg attcggtaat ctccgaacag aaggaagaac gaaggaagga gcacagactt
     4861 agattggtat atatacgcat atgtagtgtt gaagaaacat gaaattgccc agtattctta
     4921 acccaactgc acagaacaaa aacctgcagg aaacgaagat aaatcatgtc gaaagctaca
     4981 tataaggaac gtgctgctac tcatcctagt cctgttgctg ccaagctatt taatatcatg
     5041 cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc gtaccaccaa ggaattactg
     5101 gagttagttg aagcattagg tcccaaaatt tgtttactaa aaacacatgt ggatatcttg
     5161 actgattttt ccatggaggg cacagttaag ccgctaaagg cattatccgc caagtacaat
     5221 tttttactct tcgaagacag aaaatttgct gacattggta atacagtcaa attgcagtac
     5281 tctgcgggtg tatacagaat agcagaatgg gcagacatta cgaatgcaca cggtgtggtg
     5341 ggcccaggta ttgttagcgg tttgaagcag gcggcagaag aagtaacaaa ggaacctaga
     5401 ggccttttga tgttagcaga attgtcatgc aagggctccc tatctactgg agaatatact
     5461 aagggtactg ttgacattgc gaagagcgac aaagattttg ttatcggctt tattgctcaa
     5521 agagacatgg gtggaagaga tgaaggttac gattggttga ttatgacacc cggtgtgggt
     5581 ttagatgaca agggagacgc attgggtcaa cagtatagaa ccgtggatga tgtggtctct
     5641 acaggatctg acattattat tgttggaaga ggactatttg caaagggaag ggatgctaag
     5701 gtagagggtg aacgttacag aaaagcaggc tgggaagcat atttgagaag atgcggccag
     5761 caaaactaaa aaactgtatt ataagtaaat gcatgtatac taaactcaca aattagagct
     5821 tcaatttaat tatatcagtt attaccctat gcggtgtgaa ataccgcaca gatgcgtaag
     5881 gagaaaatac cgcatcagga aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     5941 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     6001 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     6061 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     6121 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     6181 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     6241 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     6301 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtccatt
     6361 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     6421 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     6481 cccagtcacg acgtt
//
