LOCUS       sequence-229432-        7605 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of CURT_fluoB controlled by the Gal1 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7605)
  AUTHORS   Behrendorff JBYH, Sandoval-Ibanez OA, Sharma A, Pribil M
  TITLE     Membrane-bound protein scaffolding in diverse hosts using thylakoid 
            protein CURT1A.
  JOURNAL   ACS Synth Biol. 2019 Mar 18. doi: 10.1021/acssynbio.8b00418.
  PUBMED    30884945
REFERENCE   2  (bases 1 to 7605)
  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: "ACS Synth 
            Biol. 2019 Mar 18. doi: 10.1021/acssynbio.8b00418."
COMMENT     Sequence Label: pESC/CURT_fluoB
FEATURES             Location/Qualifiers
     source          1..7605
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..665
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    217..334
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     629..652
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             join(1003..1005,1006..1719)
                     /codon_start=1
                     /product="enhanced YFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1045..1066)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1306..1325)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1653..1674
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      1752..1941
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2013..2030)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2184..2772)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2264..2283)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2943..3734,3735..3803))
                     /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     3566..3585
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3804..3908)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3935..5277
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(4898..4945)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     5318..5336
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5374..5396)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5496..5515
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5539..5754
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             5755..6558
                     /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(6689..7144)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6826..6847)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7038..7057
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(7223..7388)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     7309..7333
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(7536..7559)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
ORIGIN
        1 tttcaaaaat tcttactttt tttttggatg gacgcaaaga agtttaataa tcatattaca
       61 tggcattacc accatataca tatccatata catatccata tctaatctta cttatatgtt
      121 gtggaaatgt aaagagcccc attatcttag cctaaaaaaa ccttctcttt ggaactttca
      181 gtaatacgct taactgctca ttgctatatt gaagtacgga ttagaagccg ccgagcgggt
      241 gacagccctc cgaaggaaga ctctcctccg tgcgtcctcg tcttcaccgg tcgcgttcct
      301 gaaacgcaga tgtgcctcgc gccgcactgc tccgaacaat aaagattcta caatactagc
      361 ttttatggtt atgaagagga aaaattggca gtaacctggc cccacaaacc ttcaaatgaa
      421 cgaatcaaat taacaaccat aggatgataa tgcgattagt tttttagcct tatttctggg
      481 gtaattaatc agcgaagcga tgatttttga tctattaaca gatatataaa tgcaaaaact
      541 gcataaccac tttaactaat actttcaaca ttttcggttt gtattacttc ttattcaaat
      601 gtaataaaag tatcaacaaa aaattgttaa tatacctcta tactttaacg tcaaggagaa
      661 aaaaccccgg atccctcaat ggcttcttca gaagagacct cgtccattga taccaacgaa
      721 ctcatcacag atttgaagga aaagtgggat ggtcttgaga acaaatctac tgtacttata
      781 tatggaggag gagccattgt tgctgtttgg ttatcttcca ttgttgttgg tgccatcaac
      841 tctgttcctc tgcttccgaa agttatggaa cttgtcggac tagggtacac tggatggttt
      901 gtctacagat accttctctt caagtcaagc agaaaggaat tggctgagga tattgaatcc
      961 ttgaagaaga agatcgcagg aagcgaagga ggtggtggat ctgtgagcaa gggcgaggag
     1021 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     1081 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     1141 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cttcggctac
     1201 ggcctgatgt gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     1261 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     1321 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     1381 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     1441 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     1501 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     1561 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccaag
     1621 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     1681 gccgggatca ctctcggcat ggacgagctg tacaagtaac tcgagtaagc ttggtaccgc
     1741 ggctagctaa gatccgctct aaccgaaaag gaaggagtta gacaacctga agtctaggtc
     1801 cctatttatt tttttatagt tatgttagta ttaagaacgt tatttatatt tcaaattttt
     1861 cttttttttc tgtacagacg cgtgtacgca tgtaacatta tactgaaaac cttgcttgag
     1921 aaggttttgg gacgctcgaa gatccagctg cattaatgaa tcggccaacg cgcggggaga
     1981 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     2041 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     2101 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     2161 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     2221 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     2281 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     2341 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     2401 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     2461 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     2521 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     2581 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     2641 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     2701 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     2761 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     2821 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     2881 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     2941 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     3001 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     3061 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     3121 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     3181 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     3241 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     3301 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     3361 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     3421 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     3481 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     3541 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     3601 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     3661 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     3721 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     3781 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     3841 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     3901 gttccgcgca catttccccg aaaagtgcca cctgaacgaa gcatctgtgc ttcattttgt
     3961 agaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca aagaatctga gctgcatttt
     4021 tacagaacag aaatgcaacg cgaaagcgct attttaccaa cgaagaatct gtgcttcatt
     4081 tttgtaaaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc
     4141 atttttacag aacagaaatg caacgcgaga gcgctatttt accaacaaag aatctatact
     4201 tcttttttgt tctacaaaaa tgcatcccga gagcgctatt tttctaacaa agcatcttag
     4261 attacttttt ttctcctttg tgcgctctat aatgcagtct cttgataact ttttgcactg
     4321 taggtccgtt aaggttagaa gaaggctact ttggtgtcta ttttctcttc cataaaaaaa
     4381 gcctgactcc acttcccgcg tttactgatt actagcgaag ctgcgggtgc attttttcaa
     4441 gataaaggca tccccgatta tattctatac cgatgtggat tgcgcatact ttgtgaacag
     4501 aaagtgatag cgttgatgat tcttcattgg tcagaaaatt atgaacggtt tcttctattt
     4561 tgtctctata tactacgtat aggaaatgtt tacattttcg tattgttttc gattcactct
     4621 atgaatagtt cttactacaa tttttttgtc taaagagtaa tactagagat aaacataaaa
     4681 aatgtagagg tcgagtttag atgcaagttc aaggagcgaa aggtggatgg gtaggttata
     4741 tagggatata gcacagagat atatagcaaa gagatacttt tgagcaatgt ttgtggaagc
     4801 ggtattcgca atattttagt agctcgttac agtccggtgc gtttttggtt ttttgaaagt
     4861 gcgtcttcag agcgcttttg gttttcaaaa gcgctctgaa gttcctatac tttctagaga
     4921 ataggaactt cggaatagga acttcaaagc gtttccgaaa acgagcgctt ccgaaaatgc
     4981 aacgcgagct gcgcacatac agctcactgt tcacgtcgca cctatatctg cgtgttgcct
     5041 gtatatatat atacatgaga agaacggcat agtgcgtgtt tatgcttaaa tgcgtactta
     5101 tatgcgtcta tttatgtagg atgaaaggta gtctagtacc tcctgtgata ttatcccatt
     5161 ccatgcgggg tatcgtatgc ttccttcagc actacccttt agctgttcta tatgctgcca
     5221 ctcctcaatt ggattagtct catccttcaa tgctatcatt tcctttgata ttggatcata
     5281 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     5341 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     5401 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     5461 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
     5521 agtgcaccat accacctttt caattcatca tttttttttt attctttttt ttgatttcgg
     5581 tttccttgaa atttttttga ttcggtaatc tccgaacaga aggaagaacg aaggaaggag
     5641 cacagactta gattggtata tatacgcata tgtagtgttg aagaaacatg aaattgccca
     5701 gtattcttaa cccaactgca cagaacaaaa acctgcagga aacgaagata aatcatgtcg
     5761 aaagctacat ataaggaacg tgctgctact catcctagtc ctgttgctgc caagctattt
     5821 aatatcatgc acgaaaagca aacaaacttg tgtgcttcat tggatgttcg taccaccaag
     5881 gaattactgg agttagttga agcattaggt cccaaaattt gtttactaaa aacacatgtg
     5941 gatatcttga ctgatttttc catggagggc acagttaagc cgctaaaggc attatccgcc
     6001 aagtacaatt ttttactctt cgaagacaga aaatttgctg acattggtaa tacagtcaaa
     6061 ttgcagtact ctgcgggtgt atacagaata gcagaatggg cagacattac gaatgcacac
     6121 ggtgtggtgg gcccaggtat tgttagcggt ttgaagcagg cggcagaaga agtaacaaag
     6181 gaacctagag gccttttgat gttagcagaa ttgtcatgca agggctccct atctactgga
     6241 gaatatacta agggtactgt tgacattgcg aagagcgaca aagattttgt tatcggcttt
     6301 attgctcaaa gagacatggg tggaagagat gaaggttacg attggttgat tatgacaccc
     6361 ggtgtgggtt tagatgacaa gggagacgca ttgggtcaac agtatagaac cgtggatgat
     6421 gtggtctcta caggatctga cattattatt gttggaagag gactatttgc aaagggaagg
     6481 gatgctaagg tagagggtga acgttacaga aaagcaggct gggaagcata tttgagaaga
     6541 tgcggccagc aaaactaaaa aactgtatta taagtaaatg catgtatact aaactcacaa
     6601 attagagctt caatttaatt atatcagtta ttaccctatg cggtgtgaaa taccgcacag
     6661 atgcgtaagg agaaaatacc gcatcaggaa attgtaagcg ttaatatttt gttaaaattc
     6721 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     6781 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     6841 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     6901 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     6961 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     7021 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     7081 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     7141 gcgtccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     7201 cgctattacg ccagctgaat tggagcgacc tcatgctata cctgagaaag caacctgacc
     7261 tacaggaaag agttactcaa gaataagaat tttcgtttta aaacctaaga gtcactttaa
     7321 aatttgtata cacttatttt ttttataact tatttaataa taaaaatcat aaatcataag
     7381 aaattcgctt atttagaagt gtcaacaacg tatctaccaa cgatttgacc cttttccatc
     7441 ttttcgtaaa tttctggcaa ggtagacaag ccgacaacct tgattggaga cttgaccaaa
     7501 cctctggcga agaattgtta attaagagct cagatcttat cgtcgtcatc cttgtaatcc
     7561 atcgatacta gtgcggccgc cctttagtga gggttgaatt cgaat
//
