LOCUS       sequence-446738-        5631 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing StayGold under the control of the TEF1 promtoer in 
            Saccharomyces cerevisiae.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5631)
  AUTHORS   Evans S, Tominaga M, Lu Z, Bandari NC, McDonnell L, Wang C, Speight 
            RE, Ishii J, Vickers CE, Peng B
  TITLE     Characterization of Fluorescent Reporters for Flow Cytometry-Based 
            Single-Cell Studies in Saccharomyces cerevisiae.
  JOURNAL   ACS Synth Biol. 2025 Sep 19;14(9):3804-3812. doi: 
            10.1021/acssynbio.5c00432. Epub 2025 Sep 10.
  PUBMED    40927980
REFERENCE   2  (bases 1 to 5631)
  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: 
            "10.1021/acssynbio.5c00432"; journalName: "ACS Synth Biol"; date: 
            "2025-09-19- 19"; volume: "14"; issue: "9"; pages: "3804-3812"
COMMENT     Sequence Label: pILFPQ13B7
FEATURES             Location/Qualifiers
     source          1..5631
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             33..683
                     /codon_start=1
                     /product="bright and highly photostable dimeric green 
                     fluorescent protein (Hirano et al., 2022)"
                     /label=StayGold
                     /note="engineered variant of a GFP from Cytaeis uchidae"
                     /translation="MASTPFKFQLKGTINGKSFTVEGEGEGNSHEGSHKGKYVCTSGKL
                     PMSWAALGTSFGYGMKYYTKYPSGLKNWFHEVMPEGFTYDRHIQYKGDGSIHAKHQHFM
                     KNGTYHNIVEFTGQDFKENSPVLTGDMNVSLPNEVQHIPRDDGVECPVTLLYPLLSDKS
                     KCVEAHQNTICKPLHNQPAPDVPYHWIRKQYTQSKDDTEERDHICQSETLEAHL"
     terminator      701..971
                     /gene="S. cerevisae PGK1"
                     /label=PGK1 terminator
                     /note="transcription terminator for 3-phosphoglycerate 
                     kinase gene"
     primer_bind     complement(1236..1252)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1236..1252)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1249..1271)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1260..1276
                     /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(1284..1290,1291..1308,1309..1314))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1329..1350
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1467..1484)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1638..2226)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1718..1737)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2397..3188,3189..3257))
                     /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     3020..3039
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3258..3362)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3430..3448
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3486..3508)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3608..3627
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3821..3843
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3835..3852
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3836..3852
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4966..4999)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        join(5219..5631,1)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
ORIGIN
        1 gttttaatta caaactcgag aaaggatcca aaatggcttc tactccattc aagtttcagt
       61 tgaagggtac tattaacggc aagtctttta ccgttgaagg tgaaggtgag ggtaattctc
      121 atgaaggttc tcataagggc aaatacgtat gtacttctgg caaattgcca atgtcttggg
      181 ctgctttggg tacttctttt ggttatggta tgaagtatta cacgaagtac ccctctggtt
      241 tgaagaattg gtttcatgaa gttatgcctg agggtttcac ttacgataga catattcagt
      301 acaaaggtga cggttccatt catgccaaac atcaacattt tatgaagaac ggcacctacc
      361 ataacatcgt tgagtttact ggtcaggact tcaaagaaaa ctctccagtt ttgactggtg
      421 acatgaatgt ttctttgcca aacgaagttc aacacatccc aagagatgat ggtgttgaat
      481 gtccagttac tttgttgtac ccactgttgt ctgataagtc taagtgtgtt gaagctcacc
      541 aaaacactat ctgtaagcca ttgcataatc aaccagctcc agatgttcca taccattgga
      601 ttagaaaaca gtacacccaa tccaaggatg acactgaaga aagagatcat atctgccaat
      661 ctgaaacctt ggaagctcat ttgtaaagat ctgatcttaa attgaattga attgaaatcg
      721 atagatcaat ttttttcttt tctctttccc catcctttac gctaaaataa tagtttattt
      781 tattttttga atatttttta tttatatacg tatatataga ctattattta tcttttaatg
      841 attattaaga tttttattaa aaaaaaattc gctcctcttt taatgccttt atgcagtttt
      901 tttttcccat tcgatatttc tatgttcggg ttcagcgtat tttaagttta ataactcgaa
      961 aattctgcgt tcgttaaagc tttcgagaag gatattattt actagtagga ctatttgcaa
     1021 agggaaggga tgctaaggta gagggtgaac gttacagaaa agcaggctgg gaagcatatt
     1081 tgagaagatg cggccagcaa aactaaaaaa ctgtattata agtaaatgca tgtatactaa
     1141 actcacaaat tagagcttca atttaattat atcagttatt acccgggaat ctcggtcgta
     1201 atgatttaaa tgcatgcaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     1261 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     1321 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     1381 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     1441 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     1501 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     1561 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     1621 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     1681 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     1741 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     1801 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     1861 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     1921 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     1981 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     2041 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     2101 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     2161 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     2221 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     2281 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     2341 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     2401 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     2461 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     2521 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     2581 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     2641 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     2701 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     2761 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     2821 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     2881 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     2941 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     3001 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     3061 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     3121 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     3181 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     3241 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     3301 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     3361 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     3421 acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt
     3481 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     3541 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
     3601 aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg
     3661 cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac
     3721 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
     3781 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa
     3841 acgacggcca gtgaattcat ttaaattttt ttgattcggt aatctccgaa cagaaggaag
     3901 aacgaaggaa ggagcacaga cttagattgg tatatatacg catatgtagt gttgaagaaa
     3961 catgaaattg cccagtattc ttaacccaac tgcacagaac aaaaacctgc aggaaacgaa
     4021 gataaaacat gtccacaaaa tcatatacca gtagagctga gactcatgca agtccggttg
     4081 catcgaaact tttacgttta atggatgaaa agaagaccaa tttgtgtgct tctcttgacg
     4141 ttcgttcgac tgatgagcta ttgaaacttg ttgaaacgtt gggtccatac atttgccttt
     4201 tgaaaacaca cgttgatatc ttggatgatt tcagttatga gggtactgtc gttccattga
     4261 aagcattggc agagaaatac aagttcttga tatttgagga cagaaaattc gccgatatcg
     4321 gtaacacagt caaattacaa tatacatcgg gcgtttaccg tatcgcagaa tggtctgata
     4381 tcaccaacgc ccacggggtt actggtgctg gtattgttgc tggcttgaaa caaggtgcgc
     4441 aagaggtcac caaagaacca aggggattat tgatgcttgc tgaattgtct tccaagggtt
     4501 ctctagcaca cggtgaatat actaagggta ccgttgatat tgcaaagagt gataaagatt
     4561 tcgttattgg gttcattgct cagaacgata tgggaggaag agaagaaggg tttgattggc
     4621 taatcatgac cccaggtgta ggtttagacg acaaaggcga tgcattgggt cagcagtaca
     4681 gaaccgtcga cgaagttgta agtggtggat cagatatcat cattgttggc agaggacttt
     4741 tcgccaaggg tagagatcct aaggttgaag gtgaaagata cagaaatgct ggatgggaag
     4801 cgtaccaaaa gagaatcagc gctccccatt aattatacag gaaacttaat agaacaaatc
     4861 acatatttaa tctaatagcc acctgcattg gcacggtgca acactacttc aacttcatct
     4921 tacaaaaaga tcacgtgatc tgttgtattg ggatctctag acctaataac ttcgtatagc
     4981 atacattata cgaagttata ttaagggttg ctcgagaaag agctccagaa agcgaccacc
     5041 caactttggc tgataatagc gtataaacaa tgcatacttt gtacgttcaa aatacaatgc
     5101 agtagatata tttatgcata ttacatataa tacatatcac ataggaagca acaggcgcgt
     5161 tggactttta attttcgagg accgcgaatc cttacatcac acccaatccc ccacaagtga
     5221 tcccccacac accatagctt caaaatgttt ctactccttt tttactcttc cagattttct
     5281 cggactccgc gcatcgccgt accacttcaa aacacccaag cacagcatac taaatttccc
     5341 ctctttcttc ctctagggtg tcgttaatta cccgtactaa aggtttggaa aagaaaaaag
     5401 agaccgcctc gtttcttttt cttcgtcgaa aaaggcaata aaaattttta tcacgtttct
     5461 ttttcttgaa aatttttttt tttgattttt ttctctttcg atgacctccc attgatattt
     5521 aagttaataa acggtcttca atttctcaag tttcagtttc atttttcttg ttctattaca
     5581 acttttttta cttcttgctc attagaaaga aagcatagca atctaatcta a
//
