LOCUS       sequence-189855-        5929 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reporter plasmid for the PhiReX system. Encodes the reporter gene 
            yEGFP under the control of the CYC1 minimal promoter with upstream 
            synTALE-DBS..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5929)
  AUTHORS   Hochrein L, Machens F, Messerschmidt K, Mueller-Roeber B
  TITLE     PhiReX: a programmable and red light-regulated protein expression 
            switch for yeast.
  JOURNAL   Nucleic Acids Res. 2017 Sep 6;45(15):9193-9205. doi: 
            10.1093/nar/gkx610.
  PUBMED    28911120
REFERENCE   2  (bases 1 to 5929)
  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"; doi: "10"; journalName: 
            "Nucleic Acids Res"; date: "2017-09-6- 6"; volume: "45"; issue: 
            "15"; pages: "9193-9205"
COMMENT     Sequence Label: pRL_yEGFP_hc
FEATURES             Location/Qualifiers
     source          1..5929
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      61..649
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     550..569
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(774..1577)
                     /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"
     promoter        complement(1578..1793)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      2394..3271
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     primer_bind     3442..3459
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(3617..4411)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(3617..4411)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3728..3747)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4338..4357
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4412..4516)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4917..5633
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      5635..5882
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5651..5669)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
ORIGIN
        1 aatggcgcgc cgtttaaacc ctgcaggggc cggccccgta gaaaagatca aaggatcttc
       61 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
      121 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      181 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      241 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      301 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      361 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      421 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      481 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      541 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      601 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      661 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcatgata taattaaatt
      721 gaagctctaa tttgtgagtt tagtatacat gcatttactt ataatacagt tttttagttt
      781 tgctggccgc atcttctcaa atatgcttcc cagcctgctt ttctgtaacg ttcaccctct
      841 accttagcat cccttccctt tgcaaatagt cctcttccaa caataataat gtcagatcct
      901 gtagagacca catcatccac ggttctatac tgttgaccca atgcgtctcc cttgtcatct
      961 aaacccacac cgggtgtcat aatcaaccaa tcgtaacctt catctcttcc acccatgtct
     1021 ctttgagcaa taaagccgat aacaaaatct ttgtcgctct tcgcaatgtc aacagtaccc
     1081 ttagtatatt ctccagtaga tagggagccc ttgcatgaca attctgctaa catcaaaagg
     1141 cctctaggtt cctttgttac ttcttctgcc gcctgcttca aaccgctaac aatacctggg
     1201 cccaccacac cgtgtgcatt cgtaatgtct gcccattctg ctattctgta tacacccgca
     1261 gagtactgca atttgactgt attaccaatg tcagcaaatt ttctgtcttc gaagagtaaa
     1321 aaattgtact tggcggataa tgcctttagc ggcttaactg tgccctccat ggaaaaatca
     1381 gtcaagatat ccacatgtgt ttttagtaaa caaattttgg gacctaatgc ttcaactaac
     1441 tccagtaatt ccttggtggt acgaacatcc aatgaagcac acaagtttgt ttgcttttcg
     1501 tgcatgatat taaatagctt ggcagcaaca ggactaggat gagtagcagc acgttcctta
     1561 tatgtagctt tcgacatgat ttatcttcgt ttcctgcagg tttttgttct gtgcagttgg
     1621 gttaagaata ctgggcaatt tcatgtttct tcaacactac atatgcgtat atataccaat
     1681 ctaagtctgt gctccttcct tcgttcttcc ttctgttcgg agattaccga atcaaaaaaa
     1741 tttcaaggaa accgaaatca aaaaaaagaa taaaaaaaaa atgatgaatt gaaaagctag
     1801 cttatcgatg ataagctgtc aaacatgaga attaattcca cggactatag actataccta
     1861 gtatactccg tctactgtac gatacacttc cgctcaggtc cttgtccttt aacgaggcct
     1921 taccactctt ttgttactct attgatccag ctcagcaaag gcagtgtgat ctaagattct
     1981 atcttcgcga tgtagtaaaa ctagctagac cgagaaagag actagaaatg caaaaggcac
     2041 ttctacaatg gctgccatca ttattatccg atgtgacgct gcagcttctc aatgatattc
     2101 gaatacgctt tgaggagata cagcctaata tccgacaaac tgttttacag atttacgatc
     2161 gtacttgtta cccatcattg aattttgaac atccgaacct gggagttttc cctgaaacag
     2221 atagtatatt tgaacctgta taataatata tagtctagcg ctttacggaa gacaatgtat
     2281 gtatttcggt tcctggagaa actattgcat ctattgcata ggtaatcttg cacgtcgcat
     2341 ccccggttca ttttctgcgt ttccatcttg cacttcaata gcatatcttt gttaacgaag
     2401 catctgtgct tcattttgta gaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa
     2461 agaatctgag ctgcattttt acagaacaga aatgcaacgc gaaagcgcta ttttaccaac
     2521 gaagaatctg tgcttcattt ttgtaaaaca aaaatgcaac gcgagagcgc taatttttca
     2581 aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgagag cgctatttta
     2641 ccaacaaaga atctatactt cttttttgtt ctacaaaaat gcatcccgag agcgctattt
     2701 ttctaacaaa gcatcttaga ttactttttt tctcctttgt gcgctctata atgcagtctc
     2761 ttgataactt tttgcactgt aggtccgtta aggttagaag aaggctactt tggtgtctat
     2821 tttctcttcc ataaaaaaag cctgactcca cttcccgcgt ttactgatta ctagcgaagc
     2881 tgcgggtgca ttttttcaag ataaaggcat ccccgattat attctatacc gatgtggatt
     2941 gcgcatactt tgtgaacaga aagtgatagc gttgatgatt cttcattggt cagaaaatta
     3001 tgaacggttt cttctatttt gtctctatat actacgtata ggaaatgttt acattttcgt
     3061 attgttttcg attcactcta tgaatagttc ttactacaat ttttttgtct aaagagtaat
     3121 actagagata aacataaaaa atgtagaggt cgagtttaga tgcaagttca aggagcgaaa
     3181 ggtggatggg taggttatat agggatatag cacagagata tatagcaaag agatactttt
     3241 gagcaatgtt tgtggaagcg gtattcgcaa tgggaagctc caccccggtt gataatcaga
     3301 aaagccccaa aaacaggaag attgtataag caaatattta aattgtaatt ctttcctgcg
     3361 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     3421 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gggtctgacg
     3481 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     3541 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     3601 aaacttggtc tgacagtcag aagaactcgt caagaaggcg atagaaggcg atgcgctgcg
     3661 aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg ccgccaagct
     3721 cttcagcaat atcacgggta gccaacgcta tgtcctgata gcggtccgcc acacccagcc
     3781 ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc ggcaagcagg
     3841 catcgccatg agtcacgacg agatcctcgc cgtcgggcat gctcgccttg agcctggcga
     3901 acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga tcgacaagac
     3961 cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc
     4021 aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg gatactttct
     4081 cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc aatagcagcc
     4141 agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg
     4201 ccagccacga tagccgcgct gcctcgtctt gcagttcatt cagggcaccg gacaggtcgg
     4261 tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg gcatcagagc
     4321 agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa gcggccggag
     4381 aacctgcgtg caatccatct tgttcaatca tactcttcct ttttcaatat tattgaagca
     4441 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     4501 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgga cggatcgctt
     4561 gcctgtaact tacacgcgcc tcgtggcgcg cccctgcagg atttaaatgg ccggccgttt
     4621 aaactcgctg ttttcgaaat taccctttaa gcgcgggtat tggcgcgcct ctataagatc
     4681 ttgtgtgcgc tagctctata agatcttgtg tgcgctagct ctataagatc ttgtgtgcgc
     4741 tagctctata agatcttgtg tgccctaggg catgtgctct gtatgtatat aaaactcttg
     4801 ttttcttctt ttctctaaat attctttcct tatacattag gacctttgca gcataaatta
     4861 ctatacttct atagacacac aaacacaaat acacacacta aattaatagc ggccgcatgt
     4921 ctaaaggtga agaattattc actggtgttg tcccaatttt ggttgaatta gatggtgatg
     4981 ttaatggtca caaattttct gtctccggtg aaggtgaagg tgatgctact tacggtaaat
     5041 tgaccttaaa atttatttgt actactggta aattgccagt tccatggcca accttagtca
     5101 ctactttcgg ttatggtgtt caatgttttg cgagataccc agatcatatg aaacaacatg
     5161 actttttcaa gtctgccatg ccagaaggtt atgttcaaga aagaactatt tttttcaaag
     5221 atgacggtaa ctacaagacc agagctgaag tcaagtttga aggtgatacc ttagttaata
     5281 gaatcgaatt aaaaggtatt gattttaaag aagatggtaa cattttaggt cacaaattgg
     5341 aatacaacta taactctcac aatgtttaca tcatggctga caaacaaaag aatggtatca
     5401 aagttaactt caaaattaga cacaacattg aagatggttc tgttcaatta gctgaccatt
     5461 atcaacaaaa tactccaatt ggtgatggtc cagtcttgtt accagacaac cattacttat
     5521 ccactcaatc tgccttatcc aaagatccaa acgaaaagag agaccacatg gtcttgttag
     5581 aatttgttac tgctgctggt attacccatg gtatggatga attgtacaaa taattaatta
     5641 aattagttat gtcacgctta cattcacgcc ctccccccac atccgctcta accgaaaagg
     5701 aaggagttag acaacctgaa gtctaggtcc ctatttattt ttttatagtt atgttagtat
     5761 taagaacgtt atttatattt caaatttttc ttttttttct gtacagacgc gtgtacgcat
     5821 gtaacattat actgaaaacc ttgcttgaga aggttttggg acgctcgaag gctttaattt
     5881 gcaagccgaa ccaggcttat gcccagcatc gttgcaagca gacgattta
//
