LOCUS       sequence-247323-        6507 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Reporter plasmid, pZF(4BSc)-yEVENUS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6507)
  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 6507)
  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..6507
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        196..416
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             417..1220
                     /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(1351..1806)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1488..1509)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1700..1719
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1936..1958
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1950..1967
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1951..1967
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1977..1996
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1977..1995
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     2666..2689
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     2708..2724
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2709..2725
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2758..3474
                     /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      3498..3685
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3553..3577)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(3733..3753)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3733..3751)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3772..3788)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3772..3788)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3785..3807)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3796..3812
                     /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(3820..3826,3827..3844,3845..3850))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(4003..4020)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4174..4762)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4254..4273)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4933..5724,5725..5793))
                     /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     5556..5575
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5794..5898)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    5935..6438
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     6480..6498
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataccac agcttttcaa ttcaattcat catttttttt ttattctttt ttttgatttc
      241 ggtttctttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg
      301 agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc
      361 cagtattctt aacccaactg cacagaacaa aaacctgcag gaaacgaaga taaatcatgt
      421 cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct gccaagctat
      481 ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt cgtaccacca
      541 aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta aaaacacatg
      601 tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag gcattatccg
      661 ccaagtacaa ttttttactc ttcgaagaca gaaaatttgc tgacattggt aatacagtca
      721 aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt acgaatgcac
      781 acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa gaagtaacaa
      841 aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc ctatctactg
      901 gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt gttatcggct
      961 ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg attatgacac
     1021 ccggtgtggg tttagatgac aagggagacg cattgggtca acagtataga accgtggatg
     1081 atgtggtctc tacaggatct gacattatta ttgttggaag aggactattt gcaaagggaa
     1141 gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca tatttgagaa
     1201 gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata ctaaactcac
     1261 aaattagagc ttcaatttaa ttatatcagt tattacccta tgcggtgtga aataccgcac
     1321 agatgcgtaa ggagaaaata ccgcatcagg aaattgtaaa cgttaatatt ttgttaaaat
     1381 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     1441 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     1501 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     1561 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     1621 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     1681 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     1741 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     1801 gcgcgtcgcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     1861 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     1921 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat
     1981 acgactcact atagggcgaa ttgggtacgt tgctagcttc gtaccttata ttgaattttc
     2041 aaaaattctt actttttttt tggatggacg caaagaagtt taataatcat attacatggc
     2101 attaccacca tatacatatc catatacata tccatatcta atcttactta tatgttgtgg
     2161 aaatgtaaag agccccatta tcttagccta aaaaaacctt ctctttggaa ctttcagtaa
     2221 tacgcttaac tgctcattgc tatattgaag tgcggccgcg cgtgggcgtg cgtgggcgtg
     2281 cgtgggcgtg cgtgggcgtc tagaccgtgc gtcctcgtct tcaccggtcg cgttcctgaa
     2341 acgcagatgt gcctcgcgcc gcactgctcc gaacaataaa gattctacaa tactagcttt
     2401 tatggttatg aagaggaaaa attggcagta acctggcccc acaaaccttc aaattaacga
     2461 atcaaattaa caaccatagg atgataatgc gattagtttt ttagccttat ttctggggta
     2521 attaatcagc gaagcgatga tttttgatct attaacagat atataaatgg aaaagctgca
     2581 taaccacttt aactaatact ttcaacattt tcagtttgta ttacttctta ttcaaatgtc
     2641 ataaaagtat caacaaaaaa ttgttaatat acctctatac tttaacgtca aggagaaaaa
     2701 actatactcg aggtcgacgg tatcgataag cttgatatcg aattcctgca gcccgggatg
     2761 tctaaaggtg aagaattatt cactggtgtt gtcccaattt tggttgaatt agatggtgat
     2821 gttaatggtc acaaattttc tgtctccggt gaaggtgaag gtgatgctac ttacggtaaa
     2881 ttgaccttaa aattgatttg tactactggt aaattgccag ttccatggcc aaccttagtc
     2941 actactttag gttatggttt gcaatgtttt gctagatacc cagatcatat gaaacaacat
     3001 gactttttca agtctgccat gccagaaggt tatgttcaag aaagaactat ttttttcaaa
     3061 gatgacggta actacaagac cagagctgaa gtcaagtttg aaggtgatac cttagttaat
     3121 agaatcgaat taaaaggtat tgattttaaa gaagatggta acattttagg tcacaaattg
     3181 gaatacaact ataactctca caatgtttac atcactgctg acaaacaaaa gaatggtatc
     3241 aaagctaact tcaaaattag acacaacatt gaagatggtg gtgttcaatt agctgaccat
     3301 tatcaacaaa atactccaat tggtgatggt ccagtcttgt taccagacaa ccattactta
     3361 tcctatcaat ctgccttatc caaagatcca aacgaaaaga gagaccacat ggtcttgtta
     3421 gaatttgtta ctgctgctgg tattacccat ggtattgatg aattgtacaa ataatagggc
     3481 gcgccacttc taaataagcg aatttcttat gatttatgat ttttattatt aaataagtta
     3541 taaaaaaaat aagtgtatac aaattttaaa gtgactctta ggttttaaaa cgaaaattct
     3601 tattcttgag taactctttc ctgtaggtca ggttgctttc tcaggtatag tatgaggtcg
     3661 ctcttattga ccacacctct accggcagat ccgctaggga taacagggta atatgagctc
     3721 cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct
     3781 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacataggag ccggaagcat
     3841 aaagtgtaaa gcctggggtg cctaatgagt gaggtaactc acattaattg cgttgcgctc
     3901 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     3961 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     4021 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     4081 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4141 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4201 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4261 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4321 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4381 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4441 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4501 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4561 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     4621 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4681 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     4741 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4801 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     4861 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     4921 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4981 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5041 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5101 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5161 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5221 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5281 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5341 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5401 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5461 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5521 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5581 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     5641 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     5701 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     5761 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     5821 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     5881 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgggtcct tttcatcacg
     5941 tgctataaaa ataattataa tttaaatttt ttaatataaa tatataaatt aaaaatagaa
     6001 agtaaaaaaa gaaattaaag aaaaaatagt ttttgttttc cgaagatgta aaagactcta
     6061 gggggatcgc caacaaatac taccttttat cttgctcttc ctgctctcag gtattaatgc
     6121 cgaattgttt catcttgtct gtgtagaaga ccacacacga aaatcctgtg attttacatt
     6181 ttacttatcg ttaatcgaat gtatatctat ttaatctgct tttcttgtct aataaatata
     6241 tatgtaaagt acgctttttg ttgaaatttt ttaaaccttt gtttattttt ttttcttcat
     6301 tccgtaactc ttctaccttc tttatttact ttctaaaatc caaatacaaa acataaaaat
     6361 aaataaacac agagtaaatt cccaaattat tccatcatta aaagatacga ggcgcgtgta
     6421 agttacaggc aagcgatccg tcctaagaaa ccattattat catgacatta acctataaaa
     6481 ataggcgtat cacgaggccc tttcgtc
//
