LOCUS       sequence-259394-        6485 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reporter plasmid, pZF(3BS)-yEVENUS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6485)
  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 6485)
  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: pMM0287
FEATURES             Location/Qualifiers
     source          1..6485
                     /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     706..729
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     748..764
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     749..765
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             798..1514
                     /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      1538..1725
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1593..1617)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(1773..1793)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1773..1791)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1812..1828)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1812..1828)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1825..1847)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1836..1852
                     /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(1860..1866,1867..1884,1885..1890))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1905..1926
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2043..2060)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2214..2802)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2294..2313)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2973..3764,3765..3833))
                     /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     3596..3615
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3834..3938)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    3975..4478
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     4519..4537
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4575..4597)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4697..4716
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4740..4955
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             4956..5759
                     /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(5890..6345)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6027..6048)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6239..6258
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     join(6471..6485,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(6485,1..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 cgtgggcgag cgtgggcgag cgtgggcgtc tagaccgtgc gtcctcgtct
      361 tcaccggtcg cgttcctgaa acgcagatgt gcctcgcgcc gcactgctcc gaacaataaa
      421 gattctacaa tactagcttt tatggttatg aagaggaaaa attggcagta acctggcccc
      481 acaaaccttc aaattaacga atcaaattaa caaccatagg atgataatgc gattagtttt
      541 ttagccttat ttctggggta attaatcagc gaagcgatga tttttgatct attaacagat
      601 atataaatgg aaaagctgca taaccacttt aactaatact ttcaacattt tcagtttgta
      661 ttacttctta ttcaaatgtc ataaaagtat caacaaaaaa ttgttaatat acctctatac
      721 tttaacgtca aggagaaaaa actatactcg aggtcgacgg tatcgataag cttgatatcg
      781 aattcctgca gcccgggatg tctaaaggtg aagaattatt cactggtgtt gtcccaattt
      841 tggttgaatt agatggtgat gttaatggtc acaaattttc tgtctccggt gaaggtgaag
      901 gtgatgctac ttacggtaaa ttgaccttaa aattgatttg tactactggt aaattgccag
      961 ttccatggcc aaccttagtc actactttag gttatggttt gcaatgtttt gctagatacc
     1021 cagatcatat gaaacaacat gactttttca agtctgccat gccagaaggt tatgttcaag
     1081 aaagaactat ttttttcaaa gatgacggta actacaagac cagagctgaa gtcaagtttg
     1141 aaggtgatac cttagttaat agaatcgaat taaaaggtat tgattttaaa gaagatggta
     1201 acattttagg tcacaaattg gaatacaact ataactctca caatgtttac atcactgctg
     1261 acaaacaaaa gaatggtatc aaagctaact tcaaaattag acacaacatt gaagatggtg
     1321 gtgttcaatt agctgaccat tatcaacaaa atactccaat tggtgatggt ccagtcttgt
     1381 taccagacaa ccattactta tcctatcaat ctgccttatc caaagatcca aacgaaaaga
     1441 gagaccacat ggtcttgtta gaatttgtta ctgctgctgg tattacccat ggtattgatg
     1501 aattgtacaa ataatagggc gcgccacttc taaataagcg aatttcttat gatttatgat
     1561 ttttattatt aaataagtta taaaaaaaat aagtgtatac aaattttaaa gtgactctta
     1621 ggttttaaaa cgaaaattct tattcttgag taactctttc ctgtaggtca ggttgctttc
     1681 tcaggtatag tatgaggtcg ctcttattga ccacacctct accggcagat ccgctaggga
     1741 taacagggta atatgagctc cagcttttgt tccctttagt gagggttaat tgcgcgcttg
     1801 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     1861 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     1921 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     1981 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     2041 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     2101 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     2161 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     2221 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     2281 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     2341 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2401 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2461 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     2521 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     2581 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     2641 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     2701 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     2761 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     2821 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     2881 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     2941 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     3001 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     3061 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     3121 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     3181 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     3241 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     3301 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     3361 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     3421 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     3481 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     3541 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     3601 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     3661 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     3721 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     3781 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     3841 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     3901 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     3961 cctgggtcct tttcatcacg tgctataaaa ataattataa tttaaatttt ttaatataaa
     4021 tatataaatt aaaaatagaa agtaaaaaaa gaaattaaag aaaaaatagt ttttgttttc
     4081 cgaagatgta aaagactcta gggggatcgc caacaaatac taccttttat cttgctcttc
     4141 ctgctctcag gtattaatgc cgaattgttt catcttgtct gtgtagaaga ccacacacga
     4201 aaatcctgtg attttacatt ttacttatcg ttaatcgaat gtatatctat ttaatctgct
     4261 tttcttgtct aataaatata tatgtaaagt acgctttttg ttgaaatttt ttaaaccttt
     4321 gtttattttt ttttcttcat tccgtaactc ttctaccttc tttatttact ttctaaaatc
     4381 caaatacaaa acataaaaat aaataaacac agagtaaatt cccaaattat tccatcatta
     4441 aaagatacga ggcgcgtgta agttacaggc aagcgatccg tctaagaaac cattattatc
     4501 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
     4561 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     4621 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     4681 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca taccaccttt
     4741 tcaattcatc attttttttt tattcttttt tttgatttcg gtttccttga aatttttttg
     4801 attcggtaat ctccgaacag aaggaagaac gaaggaagga gcacagactt agattggtat
     4861 atatacgcat atgtagtgtt gaagaaacat gaaattgccc agtattctta acccaactgc
     4921 acagaacaaa aacctgcagg aaacgaagat aaatcatgtc gaaagctaca tataaggaac
     4981 gtgctgctac tcatcctagt cctgttgctg ccaagctatt taatatcatg cacgaaaagc
     5041 aaacaaactt gtgtgcttca ttggatgttc gtaccaccaa ggaattactg gagttagttg
     5101 aagcattagg tcccaaaatt tgtttactaa aaacacatgt ggatatcttg actgattttt
     5161 ccatggaggg cacagttaag ccgctaaagg cattatccgc caagtacaat tttttactct
     5221 tcgaagacag aaaatttgct gacattggta atacagtcaa attgcagtac tctgcgggtg
     5281 tatacagaat agcagaatgg gcagacatta cgaatgcaca cggtgtggtg ggcccaggta
     5341 ttgttagcgg tttgaagcag gcggcagaag aagtaacaaa ggaacctaga ggccttttga
     5401 tgttagcaga attgtcatgc aagggctccc tatctactgg agaatatact aagggtactg
     5461 ttgacattgc gaagagcgac aaagattttg ttatcggctt tattgctcaa agagacatgg
     5521 gtggaagaga tgaaggttac gattggttga ttatgacacc cggtgtgggt ttagatgaca
     5581 agggagacgc attgggtcaa cagtatagaa ccgtggatga tgtggtctct acaggatctg
     5641 acattattat tgttggaaga ggactatttg caaagggaag ggatgctaag gtagagggtg
     5701 aacgttacag aaaagcaggc tgggaagcat atttgagaag atgcggccag caaaactaaa
     5761 aaactgtatt ataagtaaat gcatgtatac taaactcaca aattagagct tcaatttaat
     5821 tatatcagtt attaccctat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     5881 cgcatcagga aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     5941 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     6001 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     6061 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     6121 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     6181 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     6241 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     6301 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtccatt cgccattcag
     6361 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     6421 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     6481 acgtt
//
