LOCUS       sequence-247318-        6477 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Reporter plasmid, pZF(1BS)-yEVENUS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6477)
  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 6477)
  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: pMM0285
FEATURES             Location/Qualifiers
     source          1..6477
                     /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     2636..2659
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     2678..2694
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2679..2695
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2728..3444
                     /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      3468..3655
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3523..3547)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(3703..3723)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3703..3721)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3742..3758)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3742..3758)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3755..3777)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3766..3782
                     /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(3790..3796,3797..3814,3815..3820))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3973..3990)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4144..4732)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4224..4243)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4903..5694,5695..5763))
                     /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     5526..5545
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5764..5868)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    5905..6408
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     6450..6468
                     /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 cgtgggcgtc tagaccgtgc
     2281 gtcctcgtct tcaccggtcg cgttcctgaa acgcagatgt gcctcgcgcc gcactgctcc
     2341 gaacaataaa gattctacaa tactagcttt tatggttatg aagaggaaaa attggcagta
     2401 acctggcccc acaaaccttc aaattaacga atcaaattaa caaccatagg atgataatgc
     2461 gattagtttt ttagccttat ttctggggta attaatcagc gaagcgatga tttttgatct
     2521 attaacagat atataaatgg aaaagctgca taaccacttt aactaatact ttcaacattt
     2581 tcagtttgta ttacttctta ttcaaatgtc ataaaagtat caacaaaaaa ttgttaatat
     2641 acctctatac tttaacgtca aggagaaaaa actatactcg aggtcgacgg tatcgataag
     2701 cttgatatcg aattcctgca gcccgggatg tctaaaggtg aagaattatt cactggtgtt
     2761 gtcccaattt tggttgaatt agatggtgat gttaatggtc acaaattttc tgtctccggt
     2821 gaaggtgaag gtgatgctac ttacggtaaa ttgaccttaa aattgatttg tactactggt
     2881 aaattgccag ttccatggcc aaccttagtc actactttag gttatggttt gcaatgtttt
     2941 gctagatacc cagatcatat gaaacaacat gactttttca agtctgccat gccagaaggt
     3001 tatgttcaag aaagaactat ttttttcaaa gatgacggta actacaagac cagagctgaa
     3061 gtcaagtttg aaggtgatac cttagttaat agaatcgaat taaaaggtat tgattttaaa
     3121 gaagatggta acattttagg tcacaaattg gaatacaact ataactctca caatgtttac
     3181 atcactgctg acaaacaaaa gaatggtatc aaagctaact tcaaaattag acacaacatt
     3241 gaagatggtg gtgttcaatt agctgaccat tatcaacaaa atactccaat tggtgatggt
     3301 ccagtcttgt taccagacaa ccattactta tcctatcaat ctgccttatc caaagatcca
     3361 aacgaaaaga gagaccacat ggtcttgtta gaatttgtta ctgctgctgg tattacccat
     3421 ggtattgatg aattgtacaa ataatagggc gcgccacttc taaataagcg aatttcttat
     3481 gatttatgat ttttattatt aaataagtta taaaaaaaat aagtgtatac aaattttaaa
     3541 gtgactctta ggttttaaaa cgaaaattct tattcttgag taactctttc ctgtaggtca
     3601 ggttgctttc tcaggtatag tatgaggtcg ctcttattga ccacacctct accggcagat
     3661 ccgctaggga taacagggta atatgagctc cagcttttgt tccctttagt gagggttaat
     3721 tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     3781 aattccacac aacataggag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     3841 gaggtaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     3901 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     3961 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     4021 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     4081 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     4141 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     4201 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     4261 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     4321 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     4381 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     4441 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     4501 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     4561 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     4621 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4681 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4741 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     4801 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     4861 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     4921 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     4981 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     5041 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     5101 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     5161 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     5221 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     5281 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     5341 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     5401 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     5461 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     5521 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     5581 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     5641 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     5701 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     5761 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     5821 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     5881 aaaagtgcca cctgggtcct tttcatcacg tgctataaaa ataattataa tttaaatttt
     5941 ttaatataaa tatataaatt aaaaatagaa agtaaaaaaa gaaattaaag aaaaaatagt
     6001 ttttgttttc cgaagatgta aaagactcta gggggatcgc caacaaatac taccttttat
     6061 cttgctcttc ctgctctcag gtattaatgc cgaattgttt catcttgtct gtgtagaaga
     6121 ccacacacga aaatcctgtg attttacatt ttacttatcg ttaatcgaat gtatatctat
     6181 ttaatctgct tttcttgtct aataaatata tatgtaaagt acgctttttg ttgaaatttt
     6241 ttaaaccttt gtttattttt ttttcttcat tccgtaactc ttctaccttc tttatttact
     6301 ttctaaaatc caaatacaaa acataaaaat aaataaacac agagtaaatt cccaaattat
     6361 tccatcatta aaagatacga ggcgcgtgta agttacaggc aagcgatccg tcctaagaaa
     6421 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtc
//
