LOCUS       sequence-218741-        6525 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Galactose-inducible red fluorescent protein (RFP) codon optimized 
            for S. cerevisiae.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6525)
  TITLE     RFP, codon optimized for S. cerevisiae
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6525)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pNB2289
FEATURES             Location/Qualifiers
     source          1..6525
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..443
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    2..119
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     414..437
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     475..494
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        475..493
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1274..1521
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1290..1308)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1598..1615)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1769..2357)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1849..1868)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2528..3319,3320..3388))
                     /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     3151..3170
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(3484..4287)
                     /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(4288..4503)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      5104..5981
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      complement(5992..6505)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6187..6208)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6399..6418
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 acggattaga agccgccgag cgggtgacag ccctccgaag gaagactctc ctccgtgcgt
       61 cctcgtcttc accggtcgcg ttcctgaaac gcagatgtgc ctcgcgccgc actgctccga
      121 acaataaaga ttctacaata ctagctttta tggttatgaa gaggaaaaat tggcagtaac
      181 ctggccccac aaaccttcaa atgaacgaat caaattaaca accataggat gataatgcga
      241 ttagtttttt agccttattt ctggggtaat taatcagcga agcgatgatt tttgatctat
      301 taacagatat ataaatgcaa aaactgcata accactttaa ctaatacttt caacattttc
      361 ggtttgtatt acttcttatt caaatgtaat aaaagtatca acaaaaaatt gttaatatac
      421 ctctatactt taacgtcaag gagaaaaaac cccggatcgg actactagca gctgtaatac
      481 gactcactat agggaatatt aagcttatgg acaacactga agacgttatc aaggaattca
      541 tgcaattcaa ggttagaatg gaaggttctg ttaacggtca ctacttcgaa atcgagggcg
      601 aaggtgaagg taagccatac gaaggtactc aaactgctaa gttgcaagtt actaagggtg
      661 gtccattgcc attcgcttgg gacatcttgt ctccacaatt ccaatacggt tctaaggctt
      721 acgttaaaca tcctgctgac atcccagact acatgaagtt gtctttccca gaaggtttca
      781 catgggaacg ttctatgaac ttcgaagacg gtggtgtggt tgaagttcaa caagattctt
      841 ctttgcaaga cggtactttc atctacaagg ttaagttcaa gggtgttaac ttcccagctg
      901 acggtccagt tatgcaaaag aagactgctg gttgggaacc gtctaccgaa aagttgtacc
      961 cacaagatgg cgttttgaaa ggtgaaatct ctcacgcttt gaagttgaag gacggtgggc
     1021 actatacttg tgacttcaag actgtttaca aagctaagaa gccagttcaa ttgccaggta
     1081 accattacgt tgactctaag ttggacatca ctaaccacaa cgaagactac actgttgttg
     1141 aacaatacga acacgctgaa gctagacact ctggttctca ataggatcca ctagtaacgg
     1201 ccgccagtgt gctggaattc tgcagatatc catcacactg gcggccgctc gagcatgcat
     1261 ctagagggcc gcatcatgta attagttatg tcacgcttac attcacgccc tccccccaca
     1321 tccgctctaa ccgaaaagga aggagttaga caacctgaag tctaggtccc tatttatttt
     1381 tttatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg
     1441 tacagacgcg tgtacgcatg taacattata ctgaaaacct tgcttgagaa ggttttggga
     1501 cgctcgaagg ctttaatttg caagctgcgg ccctgcatta atgaatcggc caacgcgcgg
     1561 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     1621 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     1681 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     1741 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     1801 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     1861 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     1921 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     1981 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     2041 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     2101 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     2161 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     2221 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     2281 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     2341 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     2401 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     2461 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     2521 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     2581 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     2641 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     2701 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     2761 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     2821 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     2881 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     2941 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     3001 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     3061 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     3121 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     3181 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     3241 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     3301 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     3361 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatgggta ataactgata
     3421 taattaaatt gaagctctaa tttgtgagtt tagtatacat gcatttactt ataatacagt
     3481 tttttagttt tgctggccgc atcttctcaa atatgcttcc cagcctgctt ttctgtaacg
     3541 ttcaccctct accttagcat cccttccctt tgcaaatagt cctcttccaa caataataat
     3601 gtcagatcct gtagagacca catcatccac ggttctatac tgttgaccca atgcgtctcc
     3661 cttgtcatct aaacccacac cgggtgtcat aatcaaccaa tcgtaacctt catctcttcc
     3721 acccatgtct ctttgagcaa taaagccgat aacaaaatct ttgtcgctct tcgcaatgtc
     3781 aacagtaccc ttagtatatt ctccagtaga tagggagccc ttgcatgaca attctgctaa
     3841 catcaaaagg cctctaggtt cctttgttac ttcttctgcc gcctgcttca aaccgctaac
     3901 aatacctggg cccaccacac cgtgtgcatt cgtaatgtct gcccattctg ctattctgta
     3961 tacacccgca gagtactgca atttgactgt attaccaatg tcagcaaatt ttctgtcttc
     4021 gaagagtaaa aaattgtact tggcggataa tgcctttagc ggcttaactg tgccctccat
     4081 ggaaaaatca gtcaagatat ccacatgtgt ttttagtaaa caaattttgg gacctaatgc
     4141 ttcaactaac tccagtaatt ccttggtggt acgaacatcc aatgaagcac acaagtttgt
     4201 ttgcttttcg tgcatgatat taaatagctt ggcagcaaca ggactaggat gagtagcagc
     4261 acgttcctta tatgtagctt tcgacatgat ttatcttcgt ttcctgcagg tttttgttct
     4321 gtgcagttgg gttaagaata ctgggcaatt tcatgtttct tcaacactac atatgcgtat
     4381 atataccaat ctaagtctgt gctccttcct tcgttcttcc ttctgttcgg agattaccga
     4441 atcaaaaaaa tttcaaggaa accgaaatca aaaaaaagaa taaaaaaaaa atgatgaatt
     4501 gaaaagctag cttatcgatg ataagctgtc aaacatgaga attaattcca cggactatag
     4561 actataccta gtatactccg tctactgtac gatacacttc cgctcaggtc cttgtccttt
     4621 aacgaggcct taccactctt ttgttactct attgatccag ctcagcaaag gcagtgtgat
     4681 ctaagattct atcttcgcga tgtagtaaaa ctagctagac cgagaaagag actagaaatg
     4741 caaaaggcac ttctacaatg gctgccatca ttattatccg atgtgacgct gcagcttctc
     4801 aatgatattc gaatacgctt tgaggagata cagcctaata tccgacaaac tgttttacag
     4861 atttacgatc gtacttgtta cccatcattg aattttgaac atccgaacct gggagttttc
     4921 cctgaaacag atagtatatt tgaacctgta taataatata tagtctagcg ctttacggaa
     4981 gacaatgtat gtatttcggt tcctggagaa actattgcat ctattgcata ggtaatcttg
     5041 cacgtcgcat ccccggttca ttttctgcgt ttccatcttg cacttcaata gcatatcttt
     5101 gttaacgaag catctgtgct tcattttgta gaacaaaaat gcaacgcgag agcgctaatt
     5161 tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc gaaagcgcta
     5221 ttttaccaac gaagaatctg tgcttcattt ttgtaaaaca aaaatgcaac gcgagagcgc
     5281 taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgagag
     5341 cgctatttta ccaacaaaga atctatactt cttttttgtt ctacaaaaat gcatcccgag
     5401 agcgctattt ttctaacaaa gcatcttaga ttactttttt tctcctttgt gcgctctata
     5461 atgcagtctc ttgataactt tttgcactgt aggtccgtta aggttagaag aaggctactt
     5521 tggtgtctat tttctcttcc ataaaaaaag cctgactcca cttcccgcgt ttactgatta
     5581 ctagcgaagc tgcgggtgca ttttttcaag ataaaggcat ccccgattat attctatacc
     5641 gatgtggatt gcgcatactt tgtgaacaga aagtgatagc gttgatgatt cttcattggt
     5701 cagaaaatta tgaacggttt cttctatttt gtctctatat actacgtata ggaaatgttt
     5761 acattttcgt attgttttcg attcactcta tgaatagttc ttactacaat ttttttgtct
     5821 aaagagtaat actagagata aacataaaaa atgtagaggt cgagtttaga tgcaagttca
     5881 aggagcgaaa ggtggatggg taggttatat agggatatag cacagagata tatagcaaag
     5941 agatactttt gagcaatgtt tgtggaagcg gtattcgcaa tgggaagctc caccccggtt
     6001 gataatcaga aaagccccaa aaacaggaag attgtataag caaatattta aattgtaagc
     6061 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     6121 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     6181 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     6241 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     6301 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     6361 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     6421 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     6481 cttaatgcgc cgctacaggg cgcgtgggga tcgatccact agtcc
//
