LOCUS       sequence-247320-        6497 bp    DNA     linear   UNA 15-APR-2025
DEFINITION  Reporter plasmid, pZF(3BS)-yEVENUS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6497)
  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 6497)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pMM0287
FEATURES             Location/Qualifiers
     source          1..6497
                     /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     2656..2679
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     2698..2714
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2699..2715
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2748..3464
                     /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      3488..3675
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3543..3567)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(3723..3743)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3723..3741)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3762..3778)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3762..3778)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3775..3797)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3786..3802
                     /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(3810..3816,3817..3834,3835..3840))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3993..4010)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4164..4752)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4244..4263)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4923..5714,5715..5783))
                     /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     5546..5565
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5784..5888)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    5925..6428
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     6470..6488
                     /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 cgtgggcgag cgtgggcgag
     2281 cgtgggcgtc tagaccgtgc gtcctcgtct tcaccggtcg cgttcctgaa acgcagatgt
     2341 gcctcgcgcc gcactgctcc gaacaataaa gattctacaa tactagcttt tatggttatg
     2401 aagaggaaaa attggcagta acctggcccc acaaaccttc aaattaacga atcaaattaa
     2461 caaccatagg atgataatgc gattagtttt ttagccttat ttctggggta attaatcagc
     2521 gaagcgatga tttttgatct attaacagat atataaatgg aaaagctgca taaccacttt
     2581 aactaatact ttcaacattt tcagtttgta ttacttctta ttcaaatgtc ataaaagtat
     2641 caacaaaaaa ttgttaatat acctctatac tttaacgtca aggagaaaaa actatactcg
     2701 aggtcgacgg tatcgataag cttgatatcg aattcctgca gcccgggatg tctaaaggtg
     2761 aagaattatt cactggtgtt gtcccaattt tggttgaatt agatggtgat gttaatggtc
     2821 acaaattttc tgtctccggt gaaggtgaag gtgatgctac ttacggtaaa ttgaccttaa
     2881 aattgatttg tactactggt aaattgccag ttccatggcc aaccttagtc actactttag
     2941 gttatggttt gcaatgtttt gctagatacc cagatcatat gaaacaacat gactttttca
     3001 agtctgccat gccagaaggt tatgttcaag aaagaactat ttttttcaaa gatgacggta
     3061 actacaagac cagagctgaa gtcaagtttg aaggtgatac cttagttaat agaatcgaat
     3121 taaaaggtat tgattttaaa gaagatggta acattttagg tcacaaattg gaatacaact
     3181 ataactctca caatgtttac atcactgctg acaaacaaaa gaatggtatc aaagctaact
     3241 tcaaaattag acacaacatt gaagatggtg gtgttcaatt agctgaccat tatcaacaaa
     3301 atactccaat tggtgatggt ccagtcttgt taccagacaa ccattactta tcctatcaat
     3361 ctgccttatc caaagatcca aacgaaaaga gagaccacat ggtcttgtta gaatttgtta
     3421 ctgctgctgg tattacccat ggtattgatg aattgtacaa ataatagggc gcgccacttc
     3481 taaataagcg aatttcttat gatttatgat ttttattatt aaataagtta taaaaaaaat
     3541 aagtgtatac aaattttaaa gtgactctta ggttttaaaa cgaaaattct tattcttgag
     3601 taactctttc ctgtaggtca ggttgctttc tcaggtatag tatgaggtcg ctcttattga
     3661 ccacacctct accggcagat ccgctaggga taacagggta atatgagctc cagcttttgt
     3721 tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg
     3781 tgaaattgtt atccgctcac aattccacac aacataggag ccggaagcat aaagtgtaaa
     3841 gcctggggtg cctaatgagt gaggtaactc acattaattg cgttgcgctc actgcccgct
     3901 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     3961 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     4021 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4081 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4141 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4201 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4261 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4321 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     4381 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4441 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4501 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4561 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     4621 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4681 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4741 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4801 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     4861 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     4921 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     4981 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     5041 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     5101 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     5161 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     5221 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     5281 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     5341 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     5401 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     5461 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     5521 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     5581 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     5641 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     5701 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     5761 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     5821 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     5881 gttccgcgca catttccccg aaaagtgcca cctgggtcct tttcatcacg tgctataaaa
     5941 ataattataa tttaaatttt ttaatataaa tatataaatt aaaaatagaa agtaaaaaaa
     6001 gaaattaaag aaaaaatagt ttttgttttc cgaagatgta aaagactcta gggggatcgc
     6061 caacaaatac taccttttat cttgctcttc ctgctctcag gtattaatgc cgaattgttt
     6121 catcttgtct gtgtagaaga ccacacacga aaatcctgtg attttacatt ttacttatcg
     6181 ttaatcgaat gtatatctat ttaatctgct tttcttgtct aataaatata tatgtaaagt
     6241 acgctttttg ttgaaatttt ttaaaccttt gtttattttt ttttcttcat tccgtaactc
     6301 ttctaccttc tttatttact ttctaaaatc caaatacaaa acataaaaat aaataaacac
     6361 agagtaaatt cccaaattat tccatcatta aaagatacga ggcgcgtgta agttacaggc
     6421 aagcgatccg tcctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     6481 cacgaggccc tttcgtc
//
