LOCUS       sequence-259396-        6485 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reporter plasmid, pZF(3BSopp)-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: pMM0290
FEATURES             Location/Qualifiers
     source          1..6485
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     10..29
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        10..28
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     689..712
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     731..747
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     732..748
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             781..1497
                     /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      1521..1708
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1576..1600)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(1756..1776)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1756..1774)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1795..1811)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1795..1811)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1808..1830)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1819..1835
                     /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(1843..1849,1850..1867,1868..1873))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1888..1909
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2026..2043)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2197..2785)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2277..2296)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2956..3747,3748..3816))
                     /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     3579..3598
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3817..3921)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    3958..4461
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     4502..4520
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4558..4580)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4680..4699
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4723..4938
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             4939..5742
                     /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(5873..6328)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6010..6031)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6222..6241
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6454..6476
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6468..6485
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6469..6485
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gagcgcgcgt aatacgactc actatagggc gaattgggta cgttgctagc ttcgtacctt
       61 atattgaatt ttcaaaaatt cttacttttt ttttggatgg acgcaaagaa gtttaataat
      121 catattacat ggcattacca ccatatacat atccatatac atatccatat ctaatcttac
      181 ttatatgttg tggaaatgta aagagcccca ttatcttagc ctaaaaaaac cttctctttg
      241 gaactttcag taatacgctt aactgctcat tgctatattg aagtgcggcc gccgcccacg
      301 cacgcccacg cacgcccacg ctctagaccg tgcgtcctcg tcttcaccgg tcgcgttcct
      361 gaaacgcaga tgtgcctcgc gccgcactgc tccgaacaat aaagattcta caatactagc
      421 ttttatggtt atgaagagga aaaattggca gtaacctggc cccacaaacc ttcaaattaa
      481 cgaatcaaat taacaaccat aggatgataa tgcgattagt tttttagcct tatttctggg
      541 gtaattaatc agcgaagcga tgatttttga tctattaaca gatatataaa tggaaaagct
      601 gcataaccac tttaactaat actttcaaca ttttcagttt gtattacttc ttattcaaat
      661 gtcataaaag tatcaacaaa aaattgttaa tatacctcta tactttaacg tcaaggagaa
      721 aaaactatac tcgaggtcga cggtatcgat aagcttgata tcgaattcct gcagcccggg
      781 atgtctaaag gtgaagaatt attcactggt gttgtcccaa ttttggttga attagatggt
      841 gatgttaatg gtcacaaatt ttctgtctcc ggtgaaggtg aaggtgatgc tacttacggt
      901 aaattgacct taaaattgat ttgtactact ggtaaattgc cagttccatg gccaacctta
      961 gtcactactt taggttatgg tttgcaatgt tttgctagat acccagatca tatgaaacaa
     1021 catgactttt tcaagtctgc catgccagaa ggttatgttc aagaaagaac tatttttttc
     1081 aaagatgacg gtaactacaa gaccagagct gaagtcaagt ttgaaggtga taccttagtt
     1141 aatagaatcg aattaaaagg tattgatttt aaagaagatg gtaacatttt aggtcacaaa
     1201 ttggaataca actataactc tcacaatgtt tacatcactg ctgacaaaca aaagaatggt
     1261 atcaaagcta acttcaaaat tagacacaac attgaagatg gtggtgttca attagctgac
     1321 cattatcaac aaaatactcc aattggtgat ggtccagtct tgttaccaga caaccattac
     1381 ttatcctatc aatctgcctt atccaaagat ccaaacgaaa agagagacca catggtcttg
     1441 ttagaatttg ttactgctgc tggtattacc catggtattg atgaattgta caaataatag
     1501 ggcgcgccac ttctaaataa gcgaatttct tatgatttat gatttttatt attaaataag
     1561 ttataaaaaa aataagtgta tacaaatttt aaagtgactc ttaggtttta aaacgaaaat
     1621 tcttattctt gagtaactct ttcctgtagg tcaggttgct ttctcaggta tagtatgagg
     1681 tcgctcttat tgaccacacc tctaccggca gatccgctag ggataacagg gtaatatgag
     1741 ctccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata
     1801 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     1861 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     1921 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     1981 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     2041 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     2101 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     2161 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2221 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2281 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2341 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2401 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2461 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2521 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2581 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     2641 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     2701 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     2761 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     2821 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     2881 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     2941 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     3001 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     3061 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     3121 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     3181 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     3241 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     3301 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     3361 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     3421 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     3481 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     3541 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     3601 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     3661 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     3721 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     3781 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     3841 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     3901 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgggt ccttttcatc
     3961 acgtgctata aaaataatta taatttaaat tttttaatat aaatatataa attaaaaata
     4021 gaaagtaaaa aaagaaatta aagaaaaaat agtttttgtt ttccgaagat gtaaaagact
     4081 ctagggggat cgccaacaaa tactaccttt tatcttgctc ttcctgctct caggtattaa
     4141 tgccgaattg tttcatcttg tctgtgtaga agaccacaca cgaaaatcct gtgattttac
     4201 attttactta tcgttaatcg aatgtatatc tatttaatct gcttttcttg tctaataaat
     4261 atatatgtaa agtacgcttt ttgttgaaat tttttaaacc tttgtttatt tttttttctt
     4321 cattccgtaa ctcttctacc ttctttattt actttctaaa atccaaatac aaaacataaa
     4381 aataaataaa cacagagtaa attcccaaat tattccatca ttaaaagata cgaggcgcgt
     4441 gtaagttaca ggcaagcgat ccgtctaaga aaccattatt atcatgacat taacctataa
     4501 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     4561 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     4621 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     4681 ggcatcagag cagattgtac tgagagtgca ccataccacc ttttcaattc atcatttttt
     4741 ttttattctt ttttttgatt tcggtttcct tgaaattttt ttgattcggt aatctccgaa
     4801 cagaaggaag aacgaaggaa ggagcacaga cttagattgg tatatatacg catatgtagt
     4861 gttgaagaaa catgaaattg cccagtattc ttaacccaac tgcacagaac aaaaacctgc
     4921 aggaaacgaa gataaatcat gtcgaaagct acatataagg aacgtgctgc tactcatcct
     4981 agtcctgttg ctgccaagct atttaatatc atgcacgaaa agcaaacaaa cttgtgtgct
     5041 tcattggatg ttcgtaccac caaggaatta ctggagttag ttgaagcatt aggtcccaaa
     5101 atttgtttac taaaaacaca tgtggatatc ttgactgatt tttccatgga gggcacagtt
     5161 aagccgctaa aggcattatc cgccaagtac aattttttac tcttcgaaga cagaaaattt
     5221 gctgacattg gtaatacagt caaattgcag tactctgcgg gtgtatacag aatagcagaa
     5281 tgggcagaca ttacgaatgc acacggtgtg gtgggcccag gtattgttag cggtttgaag
     5341 caggcggcag aagaagtaac aaaggaacct agaggccttt tgatgttagc agaattgtca
     5401 tgcaagggct ccctatctac tggagaatat actaagggta ctgttgacat tgcgaagagc
     5461 gacaaagatt ttgttatcgg ctttattgct caaagagaca tgggtggaag agatgaaggt
     5521 tacgattggt tgattatgac acccggtgtg ggtttagatg acaagggaga cgcattgggt
     5581 caacagtata gaaccgtgga tgatgtggtc tctacaggat ctgacattat tattgttgga
     5641 agaggactat ttgcaaaggg aagggatgct aaggtagagg gtgaacgtta cagaaaagca
     5701 ggctgggaag catatttgag aagatgcggc cagcaaaact aaaaaactgt attataagta
     5761 aatgcatgta tactaaactc acaaattaga gcttcaattt aattatatca gttattaccc
     5821 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggaaattgta
     5881 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     5941 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     6001 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     6061 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     6121 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     6181 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     6241 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     6301 gcgcttaatg cgccgctaca gggcgcgtcc attcgccatt caggctgcgc aactgttggg
     6361 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
     6421 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
     6481 ccagt
//
