LOCUS       sequence-258760-        6465 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reporter plasmid, pZF(1BS)-yEVENUS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6465)
  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 6465)
  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..6465
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     660..683
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     702..718
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     703..719
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             752..1468
                     /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      1492..1679
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1547..1571)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(1727..1747)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1727..1745)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1766..1782)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1766..1782)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1779..1801)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1790..1806
                     /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(1814..1820,1821..1838,1839..1844))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1859..1880
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1997..2014)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2168..2756)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2248..2267)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2927..3718,3719..3787))
                     /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     3550..3569
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3788..3892)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    3929..4432
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     4473..4491
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4529..4551)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4651..4670
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4694..4909
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             4910..5713
                     /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(5844..6299)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5981..6002)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6193..6212
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6425..6447
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6439..6456
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6440..6456
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 taatacgact cactataggg cgaattgggt acgttgctag cttcgtacct tatattgaat
       61 tttcaaaaat tcttactttt tttttggatg gacgcaaaga agtttaataa tcatattaca
      121 tggcattacc accatataca tatccatata catatccata tctaatctta cttatatgtt
      181 gtggaaatgt aaagagcccc attatcttag cctaaaaaaa ccttctcttt ggaactttca
      241 gtaatacgct taactgctca ttgctatatt gaagtgcggc cgcgcgtggg cgtctagacc
      301 gtgcgtcctc gtcttcaccg gtcgcgttcc tgaaacgcag atgtgcctcg cgccgcactg
      361 ctccgaacaa taaagattct acaatactag cttttatggt tatgaagagg aaaaattggc
      421 agtaacctgg ccccacaaac cttcaaatta acgaatcaaa ttaacaacca taggatgata
      481 atgcgattag ttttttagcc ttatttctgg ggtaattaat cagcgaagcg atgatttttg
      541 atctattaac agatatataa atggaaaagc tgcataacca ctttaactaa tactttcaac
      601 attttcagtt tgtattactt cttattcaaa tgtcataaaa gtatcaacaa aaaattgtta
      661 atatacctct atactttaac gtcaaggaga aaaaactata ctcgaggtcg acggtatcga
      721 taagcttgat atcgaattcc tgcagcccgg gatgtctaaa ggtgaagaat tattcactgg
      781 tgttgtccca attttggttg aattagatgg tgatgttaat ggtcacaaat tttctgtctc
      841 cggtgaaggt gaaggtgatg ctacttacgg taaattgacc ttaaaattga tttgtactac
      901 tggtaaattg ccagttccat ggccaacctt agtcactact ttaggttatg gtttgcaatg
      961 ttttgctaga tacccagatc atatgaaaca acatgacttt ttcaagtctg ccatgccaga
     1021 aggttatgtt caagaaagaa ctattttttt caaagatgac ggtaactaca agaccagagc
     1081 tgaagtcaag tttgaaggtg ataccttagt taatagaatc gaattaaaag gtattgattt
     1141 taaagaagat ggtaacattt taggtcacaa attggaatac aactataact ctcacaatgt
     1201 ttacatcact gctgacaaac aaaagaatgg tatcaaagct aacttcaaaa ttagacacaa
     1261 cattgaagat ggtggtgttc aattagctga ccattatcaa caaaatactc caattggtga
     1321 tggtccagtc ttgttaccag acaaccatta cttatcctat caatctgcct tatccaaaga
     1381 tccaaacgaa aagagagacc acatggtctt gttagaattt gttactgctg ctggtattac
     1441 ccatggtatt gatgaattgt acaaataata gggcgcgcca cttctaaata agcgaatttc
     1501 ttatgattta tgatttttat tattaaataa gttataaaaa aaataagtgt atacaaattt
     1561 taaagtgact cttaggtttt aaaacgaaaa ttcttattct tgagtaactc tttcctgtag
     1621 gtcaggttgc tttctcaggt atagtatgag gtcgctctta ttgaccacac ctctaccggc
     1681 agatccgcta gggataacag ggtaatatga gctccagctt ttgttccctt tagtgagggt
     1741 taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     1801 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     1861 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     1921 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     1981 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     2041 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     2101 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     2161 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     2221 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     2281 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     2341 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     2401 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     2461 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     2521 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     2581 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     2641 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     2701 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     2761 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     2821 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     2881 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     2941 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     3001 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     3061 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     3121 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     3181 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     3241 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     3301 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     3361 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     3421 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     3481 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     3541 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     3601 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     3661 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     3721 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     3781 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     3841 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     3901 cccgaaaagt gccacctggg tccttttcat cacgtgctat aaaaataatt ataatttaaa
     3961 ttttttaata taaatatata aattaaaaat agaaagtaaa aaaagaaatt aaagaaaaaa
     4021 tagtttttgt tttccgaaga tgtaaaagac tctaggggga tcgccaacaa atactacctt
     4081 ttatcttgct cttcctgctc tcaggtatta atgccgaatt gtttcatctt gtctgtgtag
     4141 aagaccacac acgaaaatcc tgtgatttta cattttactt atcgttaatc gaatgtatat
     4201 ctatttaatc tgcttttctt gtctaataaa tatatatgta aagtacgctt tttgttgaaa
     4261 ttttttaaac ctttgtttat ttttttttct tcattccgta actcttctac cttctttatt
     4321 tactttctaa aatccaaata caaaacataa aaataaataa acacagagta aattcccaaa
     4381 ttattccatc attaaaagat acgaggcgcg tgtaagttac aggcaagcga tccgtctaag
     4441 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     4501 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     4561 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     4621 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     4681 accataccac cttttcaatt catcattttt tttttattct tttttttgat ttcggtttcc
     4741 ttgaaatttt tttgattcgg taatctccga acagaaggaa gaacgaagga aggagcacag
     4801 acttagattg gtatatatac gcatatgtag tgttgaagaa acatgaaatt gcccagtatt
     4861 cttaacccaa ctgcacagaa caaaaacctg caggaaacga agataaatca tgtcgaaagc
     4921 tacatataag gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc tatttaatat
     4981 catgcacgaa aagcaaacaa acttgtgtgc ttcattggat gttcgtacca ccaaggaatt
     5041 actggagtta gttgaagcat taggtcccaa aatttgttta ctaaaaacac atgtggatat
     5101 cttgactgat ttttccatgg agggcacagt taagccgcta aaggcattat ccgccaagta
     5161 caatttttta ctcttcgaag acagaaaatt tgctgacatt ggtaatacag tcaaattgca
     5221 gtactctgcg ggtgtataca gaatagcaga atgggcagac attacgaatg cacacggtgt
     5281 ggtgggccca ggtattgtta gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc
     5341 tagaggcctt ttgatgttag cagaattgtc atgcaagggc tccctatcta ctggagaata
     5401 tactaagggt actgttgaca ttgcgaagag cgacaaagat tttgttatcg gctttattgc
     5461 tcaaagagac atgggtggaa gagatgaagg ttacgattgg ttgattatga cacccggtgt
     5521 gggtttagat gacaagggag acgcattggg tcaacagtat agaaccgtgg atgatgtggt
     5581 ctctacagga tctgacatta ttattgttgg aagaggacta tttgcaaagg gaagggatgc
     5641 taaggtagag ggtgaacgtt acagaaaagc aggctgggaa gcatatttga gaagatgcgg
     5701 ccagcaaaac taaaaaactg tattataagt aaatgcatgt atactaaact cacaaattag
     5761 agcttcaatt taattatatc agttattacc ctatgcggtg tgaaataccg cacagatgcg
     5821 taaggagaaa ataccgcatc aggaaattgt aagcgttaat attttgttaa aattcgcgtt
     5881 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     5941 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     6001 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     6061 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     6121 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     6181 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     6241 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     6301 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     6361 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     6421 ttttcccagt cacgacgttg taaaacgacg gccagtgagc gcgcg
//
