LOCUS       sequence-229488-        7472 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of CURT_flouD controlled by the Gal1 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7472)
  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 7472)
  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_fluoD
FEATURES             Location/Qualifiers
     source          1..7472
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(159..178)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     506..527
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      605..794
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(866..883)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1037..1625)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1117..1136)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1796..2587,2588..2656))
                     /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     2419..2438
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2657..2761)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2788..4130
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(3751..3798)
                     /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     4171..4189
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4227..4249)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4349..4368
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4392..4607
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             4608..5411
                     /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(5542..5997)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5679..5700)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5891..5910
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(6076..6241)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     6162..6186
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(6389..6412)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        6459..7123
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    6675..6792
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     7087..7110
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             join(7328..7330,7331..7472,1..572)
                     /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(7370..7391)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
ORIGIN
        1 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccttcggc tacggcctga
       61 tgtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
      121 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
      181 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
      241 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
      301 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
      361 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
      421 gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc aagctgagca
      481 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
      541 tcactctcgg catggacgag ctgtacaagt aactcgagta agcttggtac cgcggctagc
      601 taagatccgc tctaaccgaa aaggaaggag ttagacaacc tgaagtctag gtccctattt
      661 atttttttat agttatgtta gtattaagaa cgttatttat atttcaaatt tttctttttt
      721 ttctgtacag acgcgtgtac gcatgtaaca ttatactgaa aaccttgctt gagaaggttt
      781 tgggacgctc gaagatccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
      841 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
      901 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
      961 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     1021 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     1081 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     1141 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     1201 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     1261 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     1321 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     1381 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     1441 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     1501 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     1561 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     1621 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     1681 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     1741 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     1801 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     1861 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     1921 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     1981 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     2041 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     2101 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     2161 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     2221 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     2281 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     2341 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     2401 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     2461 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     2521 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     2581 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     2641 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     2701 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     2761 gcacatttcc ccgaaaagtg ccacctgaac gaagcatctg tgcttcattt tgtagaacaa
     2821 aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa
     2881 cagaaatgca acgcgaaagc gctattttac caacgaagaa tctgtgcttc atttttgtaa
     2941 aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta
     3001 cagaacagaa atgcaacgcg agagcgctat tttaccaaca aagaatctat acttcttttt
     3061 tgttctacaa aaatgcatcc cgagagcgct atttttctaa caaagcatct tagattactt
     3121 tttttctcct ttgtgcgctc tataatgcag tctcttgata actttttgca ctgtaggtcc
     3181 gttaaggtta gaagaaggct actttggtgt ctattttctc ttccataaaa aaagcctgac
     3241 tccacttccc gcgtttactg attactagcg aagctgcggg tgcatttttt caagataaag
     3301 gcatccccga ttatattcta taccgatgtg gattgcgcat actttgtgaa cagaaagtga
     3361 tagcgttgat gattcttcat tggtcagaaa attatgaacg gtttcttcta ttttgtctct
     3421 atatactacg tataggaaat gtttacattt tcgtattgtt ttcgattcac tctatgaata
     3481 gttcttacta caattttttt gtctaaagag taatactaga gataaacata aaaaatgtag
     3541 aggtcgagtt tagatgcaag ttcaaggagc gaaaggtgga tgggtaggtt atatagggat
     3601 atagcacaga gatatatagc aaagagatac ttttgagcaa tgtttgtgga agcggtattc
     3661 gcaatatttt agtagctcgt tacagtccgg tgcgtttttg gttttttgaa agtgcgtctt
     3721 cagagcgctt ttggttttca aaagcgctct gaagttccta tactttctag agaataggaa
     3781 cttcggaata ggaacttcaa agcgtttccg aaaacgagcg cttccgaaaa tgcaacgcga
     3841 gctgcgcaca tacagctcac tgttcacgtc gcacctatat ctgcgtgttg cctgtatata
     3901 tatatacatg agaagaacgg catagtgcgt gtttatgctt aaatgcgtac ttatatgcgt
     3961 ctatttatgt aggatgaaag gtagtctagt acctcctgtg atattatccc attccatgcg
     4021 gggtatcgta tgcttccttc agcactaccc tttagctgtt ctatatgctg ccactcctca
     4081 attggattag tctcatcctt caatgctatc atttcctttg atattggatc atactaagaa
     4141 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     4201 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     4261 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     4321 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     4381 cataccacct tttcaattca tcattttttt tttattcttt tttttgattt cggtttcctt
     4441 gaaatttttt tgattcggta atctccgaac agaaggaaga acgaaggaag gagcacagac
     4501 ttagattggt atatatacgc atatgtagtg ttgaagaaac atgaaattgc ccagtattct
     4561 taacccaact gcacagaaca aaaacctgca ggaaacgaag ataaatcatg tcgaaagcta
     4621 catataagga acgtgctgct actcatccta gtcctgttgc tgccaagcta tttaatatca
     4681 tgcacgaaaa gcaaacaaac ttgtgtgctt cattggatgt tcgtaccacc aaggaattac
     4741 tggagttagt tgaagcatta ggtcccaaaa tttgtttact aaaaacacat gtggatatct
     4801 tgactgattt ttccatggag ggcacagtta agccgctaaa ggcattatcc gccaagtaca
     4861 attttttact cttcgaagac agaaaatttg ctgacattgg taatacagtc aaattgcagt
     4921 actctgcggg tgtatacaga atagcagaat gggcagacat tacgaatgca cacggtgtgg
     4981 tgggcccagg tattgttagc ggtttgaagc aggcggcaga agaagtaaca aaggaaccta
     5041 gaggcctttt gatgttagca gaattgtcat gcaagggctc cctatctact ggagaatata
     5101 ctaagggtac tgttgacatt gcgaagagcg acaaagattt tgttatcggc tttattgctc
     5161 aaagagacat gggtggaaga gatgaaggtt acgattggtt gattatgaca cccggtgtgg
     5221 gtttagatga caagggagac gcattgggtc aacagtatag aaccgtggat gatgtggtct
     5281 ctacaggatc tgacattatt attgttggaa gaggactatt tgcaaaggga agggatgcta
     5341 aggtagaggg tgaacgttac agaaaagcag gctgggaagc atatttgaga agatgcggcc
     5401 agcaaaacta aaaaactgta ttataagtaa atgcatgtat actaaactca caaattagag
     5461 cttcaattta attatatcag ttattaccct atgcggtgtg aaataccgca cagatgcgta
     5521 aggagaaaat accgcatcag gaaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     5581 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     5641 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     5701 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     5761 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     5821 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     5881 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     5941 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcca
     6001 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     6061 acgccagctg aattggagcg acctcatgct atacctgaga aagcaacctg acctacagga
     6121 aagagttact caagaataag aattttcgtt ttaaaaccta agagtcactt taaaatttgt
     6181 atacacttat tttttttata acttatttaa taataaaaat cataaatcat aagaaattcg
     6241 cttatttaga agtgtcaaca acgtatctac caacgatttg acccttttcc atcttttcgt
     6301 aaatttctgg caaggtagac aagccgacaa ccttgattgg agacttgacc aaacctctgg
     6361 cgaagaattg ttaattaaga gctcagatct tatcgtcgtc atccttgtaa tccatcgata
     6421 ctagtgcggc cgccctttag tgagggttga attcgaattt tcaaaaattc ttactttttt
     6481 tttggatgga cgcaaagaag tttaataatc atattacatg gcattaccac catatacata
     6541 tccatataca tatccatatc taatcttact tatatgttgt ggaaatgtaa agagccccat
     6601 tatcttagcc taaaaaaacc ttctctttgg aactttcagt aatacgctta actgctcatt
     6661 gctatattga agtacggatt agaagccgcc gagcgggtga cagccctccg aaggaagact
     6721 ctcctccgtg cgtcctcgtc ttcaccggtc gcgttcctga aacgcagatg tgcctcgcgc
     6781 cgcactgctc cgaacaataa agattctaca atactagctt ttatggttat gaagaggaaa
     6841 aattggcagt aacctggccc cacaaacctt caaatgaacg aatcaaatta acaaccatag
     6901 gatgataatg cgattagttt tttagcctta tttctggggt aattaatcag cgaagcgatg
     6961 atttttgatc tattaacaga tatataaatg caaaaactgc ataaccactt taactaatac
     7021 tttcaacatt ttcggtttgt attacttctt attcaaatgt aataaaagta tcaacaaaaa
     7081 attgttaata tacctctata ctttaacgtc aaggagaaaa aaccccggat ccctcatggg
     7141 tcttgagaac aaatctactg tacttatata tggaggagga gccattgttg ctgtttggtt
     7201 atcttccatt gttgttggtg ccatcaactc tgttcctctg cttccgaaag ttatggaact
     7261 tgtcggacta gggtacactg gatggtttgt ctacagatac cttctcttca agggaggtgg
     7321 tggatctgtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     7381 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     7441 ctacggcaag ctgaccctga agttcatctg ca
//
