LOCUS       sequence-259091-        6480 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reporter plasmid, pZF(3oBS)-yEVENUS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6480)
  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 6480)
  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: pMM0286
FEATURES             Location/Qualifiers
     source          1..6480
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(35..139)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    176..679
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     720..738
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(776..798)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     898..917
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        941..1156
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             1157..1960
                     /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(2091..2546)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2228..2249)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2440..2459
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2672..2694
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2686..2703
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2687..2703
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2713..2732
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2713..2731
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     3387..3410
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     3429..3445
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3430..3446
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3479..4195
                     /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      4219..4406
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4274..4298)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(4454..4474)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4454..4472)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4493..4509)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4493..4509)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4506..4528)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4517..4533
                     /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(4541..4547,4548..4565,4566..4571))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4586..4607
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4724..4741)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4895..5483)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4975..4994)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5654..6445,6446..6480,1..34))
                     /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     6277..6296
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
       61 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
      121 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgggtcc ttttcatcac
      181 gtgctataaa aataattata atttaaattt tttaatataa atatataaat taaaaataga
      241 aagtaaaaaa agaaattaaa gaaaaaatag tttttgtttt ccgaagatgt aaaagactct
      301 agggggatcg ccaacaaata ctacctttta tcttgctctt cctgctctca ggtattaatg
      361 ccgaattgtt tcatcttgtc tgtgtagaag accacacacg aaaatcctgt gattttacat
      421 tttacttatc gttaatcgaa tgtatatcta tttaatctgc ttttcttgtc taataaatat
      481 atatgtaaag tacgcttttt gttgaaattt tttaaacctt tgtttatttt tttttcttca
      541 ttccgtaact cttctacctt ctttatttac tttctaaaat ccaaatacaa aacataaaaa
      601 taaataaaca cagagtaaat tcccaaatta ttccatcatt aaaagatacg aggcgcgtgt
      661 aagttacagg caagcgatcc gtctaagaaa ccattattat catgacatta acctataaaa
      721 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
      781 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
      841 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
      901 catcagagca gattgtactg agagtgcacc ataccacctt ttcaattcat catttttttt
      961 ttattctttt ttttgatttc ggtttccttg aaattttttt gattcggtaa tctccgaaca
     1021 gaaggaagaa cgaaggaagg agcacagact tagattggta tatatacgca tatgtagtgt
     1081 tgaagaaaca tgaaattgcc cagtattctt aacccaactg cacagaacaa aaacctgcag
     1141 gaaacgaaga taaatcatgt cgaaagctac atataaggaa cgtgctgcta ctcatcctag
     1201 tcctgttgct gccaagctat ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc
     1261 attggatgtt cgtaccacca aggaattact ggagttagtt gaagcattag gtcccaaaat
     1321 ttgtttacta aaaacacatg tggatatctt gactgatttt tccatggagg gcacagttaa
     1381 gccgctaaag gcattatccg ccaagtacaa ttttttactc ttcgaagaca gaaaatttgc
     1441 tgacattggt aatacagtca aattgcagta ctctgcgggt gtatacagaa tagcagaatg
     1501 ggcagacatt acgaatgcac acggtgtggt gggcccaggt attgttagcg gtttgaagca
     1561 ggcggcagaa gaagtaacaa aggaacctag aggccttttg atgttagcag aattgtcatg
     1621 caagggctcc ctatctactg gagaatatac taagggtact gttgacattg cgaagagcga
     1681 caaagatttt gttatcggct ttattgctca aagagacatg ggtggaagag atgaaggtta
     1741 cgattggttg attatgacac ccggtgtggg tttagatgac aagggagacg cattgggtca
     1801 acagtataga accgtggatg atgtggtctc tacaggatct gacattatta ttgttggaag
     1861 aggactattt gcaaagggaa gggatgctaa ggtagagggt gaacgttaca gaaaagcagg
     1921 ctgggaagca tatttgagaa gatgcggcca gcaaaactaa aaaactgtat tataagtaaa
     1981 tgcatgtata ctaaactcac aaattagagc ttcaatttaa ttatatcagt tattacccta
     2041 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg aaattgtaag
     2101 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     2161 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     2221 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     2281 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     2341 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     2401 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     2461 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     2521 gcttaatgcg ccgctacagg gcgcgtccat tcgccattca ggctgcgcaa ctgttgggaa
     2581 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
     2641 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
     2701 agtgagcgcg cgtaatacga ctcactatag ggcgaattgg gtacgttgct agcttcgtac
     2761 cttatattga attttcaaaa attcttactt tttttttgga tggacgcaaa gaagtttaat
     2821 aatcatatta catggcatta ccaccatata catatccata tacatatcca tatctaatct
     2881 tacttatatg ttgtggaaat gtaaagagcc ccattatctt agcctaaaaa aaccttctct
     2941 ttggaacttt cagtaatacg cttaactgct cattgctata ttgaagtgcg gccgcgcgtg
     3001 ggcgtgggcg gcgtgggcgt ctagaccgtg cgtcctcgtc ttcaccggtc gcgttcctga
     3061 aacgcagatg tgcctcgcgc cgcactgctc cgaacaataa agattctaca atactagctt
     3121 ttatggttat gaagaggaaa aattggcagt aacctggccc cacaaacctt caaattaacg
     3181 aatcaaatta acaaccatag gatgataatg cgattagttt tttagcctta tttctggggt
     3241 aattaatcag cgaagcgatg atttttgatc tattaacaga tatataaatg gaaaagctgc
     3301 ataaccactt taactaatac tttcaacatt ttcagtttgt attacttctt attcaaatgt
     3361 cataaaagta tcaacaaaaa attgttaata tacctctata ctttaacgtc aaggagaaaa
     3421 aactatactc gaggtcgacg gtatcgataa gcttgatatc gaattcctgc agcccgggat
     3481 gtctaaaggt gaagaattat tcactggtgt tgtcccaatt ttggttgaat tagatggtga
     3541 tgttaatggt cacaaatttt ctgtctccgg tgaaggtgaa ggtgatgcta cttacggtaa
     3601 attgacctta aaattgattt gtactactgg taaattgcca gttccatggc caaccttagt
     3661 cactacttta ggttatggtt tgcaatgttt tgctagatac ccagatcata tgaaacaaca
     3721 tgactttttc aagtctgcca tgccagaagg ttatgttcaa gaaagaacta tttttttcaa
     3781 agatgacggt aactacaaga ccagagctga agtcaagttt gaaggtgata ccttagttaa
     3841 tagaatcgaa ttaaaaggta ttgattttaa agaagatggt aacattttag gtcacaaatt
     3901 ggaatacaac tataactctc acaatgttta catcactgct gacaaacaaa agaatggtat
     3961 caaagctaac ttcaaaatta gacacaacat tgaagatggt ggtgttcaat tagctgacca
     4021 ttatcaacaa aatactccaa ttggtgatgg tccagtcttg ttaccagaca accattactt
     4081 atcctatcaa tctgccttat ccaaagatcc aaacgaaaag agagaccaca tggtcttgtt
     4141 agaatttgtt actgctgctg gtattaccca tggtattgat gaattgtaca aataataggg
     4201 cgcgccactt ctaaataagc gaatttctta tgatttatga tttttattat taaataagtt
     4261 ataaaaaaaa taagtgtata caaattttaa agtgactctt aggttttaaa acgaaaattc
     4321 ttattcttga gtaactcttt cctgtaggtc aggttgcttt ctcaggtata gtatgaggtc
     4381 gctcttattg accacacctc taccggcaga tccgctaggg ataacagggt aatatgagct
     4441 ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc
     4501 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     4561 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     4621 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     4681 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     4741 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     4801 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     4861 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4921 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4981 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5041 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     5101 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5161 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5221 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5281 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     5341 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5401 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5461 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     5521 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     5581 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     5641 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     5701 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     5761 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     5821 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     5881 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     5941 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     6001 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     6061 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     6121 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     6181 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     6241 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     6301 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     6361 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     6421 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
//
