LOCUS       sequence-259395-        6485 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reporter plasmid, pZF(3BSc)-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: pMM0288
FEATURES             Location/Qualifiers
     source          1..6485
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     661..684
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     703..719
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     704..720
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             753..1469
                     /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      1493..1680
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1548..1572)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(1728..1748)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1728..1746)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1767..1783)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1767..1783)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1780..1802)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1791..1807
                     /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(1815..1821,1822..1839,1840..1845))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1860..1881
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1998..2015)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2169..2757)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2249..2268)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2928..3719,3720..3788))
                     /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     3551..3570
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3789..3893)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    3930..4433
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     4474..4492
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4530..4552)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4652..4671
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4695..4910
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             4911..5714
                     /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(5845..6300)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5982..6003)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6194..6213
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6426..6448
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6440..6457
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6441..6457
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(6467..6485,1)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6467..6485
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gcgaattggg tacgttgcta gcttcgtacc ttatattgaa ttttcaaaaa ttcttacttt
       61 ttttttggat ggacgcaaag aagtttaata atcatattac atggcattac caccatatac
      121 atatccatat acatatccat atctaatctt acttatatgt tgtggaaatg taaagagccc
      181 cattatctta gcctaaaaaa accttctctt tggaactttc agtaatacgc ttaactgctc
      241 attgctatat tgaagtgcgg ccgcgcgtgg gcgtgcgtgg gcgtgcgtgg gcgtctagac
      301 cgtgcgtcct cgtcttcacc ggtcgcgttc ctgaaacgca gatgtgcctc gcgccgcact
      361 gctccgaaca ataaagattc tacaatacta gcttttatgg ttatgaagag gaaaaattgg
      421 cagtaacctg gccccacaaa ccttcaaatt aacgaatcaa attaacaacc ataggatgat
      481 aatgcgatta gttttttagc cttatttctg gggtaattaa tcagcgaagc gatgattttt
      541 gatctattaa cagatatata aatggaaaag ctgcataacc actttaacta atactttcaa
      601 cattttcagt ttgtattact tcttattcaa atgtcataaa agtatcaaca aaaaattgtt
      661 aatatacctc tatactttaa cgtcaaggag aaaaaactat actcgaggtc gacggtatcg
      721 ataagcttga tatcgaattc ctgcagcccg ggatgtctaa aggtgaagaa ttattcactg
      781 gtgttgtccc aattttggtt gaattagatg gtgatgttaa tggtcacaaa ttttctgtct
      841 ccggtgaagg tgaaggtgat gctacttacg gtaaattgac cttaaaattg atttgtacta
      901 ctggtaaatt gccagttcca tggccaacct tagtcactac tttaggttat ggtttgcaat
      961 gttttgctag atacccagat catatgaaac aacatgactt tttcaagtct gccatgccag
     1021 aaggttatgt tcaagaaaga actatttttt tcaaagatga cggtaactac aagaccagag
     1081 ctgaagtcaa gtttgaaggt gataccttag ttaatagaat cgaattaaaa ggtattgatt
     1141 ttaaagaaga tggtaacatt ttaggtcaca aattggaata caactataac tctcacaatg
     1201 tttacatcac tgctgacaaa caaaagaatg gtatcaaagc taacttcaaa attagacaca
     1261 acattgaaga tggtggtgtt caattagctg accattatca acaaaatact ccaattggtg
     1321 atggtccagt cttgttacca gacaaccatt acttatccta tcaatctgcc ttatccaaag
     1381 atccaaacga aaagagagac cacatggtct tgttagaatt tgttactgct gctggtatta
     1441 cccatggtat tgatgaattg tacaaataat agggcgcgcc acttctaaat aagcgaattt
     1501 cttatgattt atgattttta ttattaaata agttataaaa aaaataagtg tatacaaatt
     1561 ttaaagtgac tcttaggttt taaaacgaaa attcttattc ttgagtaact ctttcctgta
     1621 ggtcaggttg ctttctcagg tatagtatga ggtcgctctt attgaccaca cctctaccgg
     1681 cagatccgct agggataaca gggtaatatg agctccagct tttgttccct ttagtgaggg
     1741 ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     1801 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     1861 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     1921 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     1981 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     2041 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     2101 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     2161 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     2221 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     2281 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     2341 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     2401 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     2461 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     2521 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     2581 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     2641 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     2701 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     2761 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     2821 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     2881 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     2941 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     3001 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     3061 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     3121 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     3181 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     3241 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     3301 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     3361 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     3421 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     3481 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     3541 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     3601 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     3661 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     3721 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     3781 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     3841 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     3901 ccccgaaaag tgccacctgg gtccttttca tcacgtgcta taaaaataat tataatttaa
     3961 attttttaat ataaatatat aaattaaaaa tagaaagtaa aaaaagaaat taaagaaaaa
     4021 atagtttttg ttttccgaag atgtaaaaga ctctaggggg atcgccaaca aatactacct
     4081 tttatcttgc tcttcctgct ctcaggtatt aatgccgaat tgtttcatct tgtctgtgta
     4141 gaagaccaca cacgaaaatc ctgtgatttt acattttact tatcgttaat cgaatgtata
     4201 tctatttaat ctgcttttct tgtctaataa atatatatgt aaagtacgct ttttgttgaa
     4261 attttttaaa cctttgttta tttttttttc ttcattccgt aactcttcta ccttctttat
     4321 ttactttcta aaatccaaat acaaaacata aaaataaata aacacagagt aaattcccaa
     4381 attattccat cattaaaaga tacgaggcgc gtgtaagtta caggcaagcg atccgtctaa
     4441 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     4501 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     4561 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     4621 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     4681 caccatacca ccttttcaat tcatcatttt ttttttattc ttttttttga tttcggtttc
     4741 cttgaaattt ttttgattcg gtaatctccg aacagaagga agaacgaagg aaggagcaca
     4801 gacttagatt ggtatatata cgcatatgta gtgttgaaga aacatgaaat tgcccagtat
     4861 tcttaaccca actgcacaga acaaaaacct gcaggaaacg aagataaatc atgtcgaaag
     4921 ctacatataa ggaacgtgct gctactcatc ctagtcctgt tgctgccaag ctatttaata
     4981 tcatgcacga aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc accaaggaat
     5041 tactggagtt agttgaagca ttaggtccca aaatttgttt actaaaaaca catgtggata
     5101 tcttgactga tttttccatg gagggcacag ttaagccgct aaaggcatta tccgccaagt
     5161 acaatttttt actcttcgaa gacagaaaat ttgctgacat tggtaataca gtcaaattgc
     5221 agtactctgc gggtgtatac agaatagcag aatgggcaga cattacgaat gcacacggtg
     5281 tggtgggccc aggtattgtt agcggtttga agcaggcggc agaagaagta acaaaggaac
     5341 ctagaggcct tttgatgtta gcagaattgt catgcaaggg ctccctatct actggagaat
     5401 atactaaggg tactgttgac attgcgaaga gcgacaaaga ttttgttatc ggctttattg
     5461 ctcaaagaga catgggtgga agagatgaag gttacgattg gttgattatg acacccggtg
     5521 tgggtttaga tgacaaggga gacgcattgg gtcaacagta tagaaccgtg gatgatgtgg
     5581 tctctacagg atctgacatt attattgttg gaagaggact atttgcaaag ggaagggatg
     5641 ctaaggtaga gggtgaacgt tacagaaaag caggctggga agcatatttg agaagatgcg
     5701 gccagcaaaa ctaaaaaact gtattataag taaatgcatg tatactaaac tcacaaatta
     5761 gagcttcaat ttaattatat cagttattac cctatgcggt gtgaaatacc gcacagatgc
     5821 gtaaggagaa aataccgcat caggaaattg taagcgttaa tattttgtta aaattcgcgt
     5881 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     5941 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     6001 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     6061 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     6121 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     6181 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     6241 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     6301 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     6361 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     6421 gttttcccag tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat acgactcact
     6481 atagg
//
