LOCUS       sequence-224422-        7612 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 7612)
  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 7612)
  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..7612
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        196..416
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             417..1220
                     /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(1351..1806)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1488..1509)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1700..1719
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(1885..2050)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     1971..1995
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(2198..2221)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        2268..2932
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    2484..2601
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     2896..2919
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             join(3270..3272,3273..3986)
                     /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(3312..3333)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3573..3592)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3920..3941
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      4019..4208
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4280..4297)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4451..5039)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4531..4550)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5210..6001,6002..6070))
                     /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     5833..5852
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6071..6175)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6202..7544
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(7165..7212)
                     /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     7585..7603
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataccac agcttttcaa ttcaattcat catttttttt ttattctttt ttttgatttc
      241 ggtttctttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg
      301 agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc
      361 cagtattctt aacccaactg cacagaacaa aaacctgcag gaaacgaaga taaatcatgt
      421 cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct gccaagctat
      481 ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt cgtaccacca
      541 aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta aaaacacatg
      601 tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag gcattatccg
      661 ccaagtacaa ttttttactc ttcgaagaca gaaaatttgc tgacattggt aatacagtca
      721 aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt acgaatgcac
      781 acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa gaagtaacaa
      841 aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc ctatctactg
      901 gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt gttatcggct
      961 ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg attatgacac
     1021 ccggtgtggg tttagatgac aagggagacg cattgggtca acagtataga accgtggatg
     1081 atgtggtctc tacaggatct gacattatta ttgttggaag aggactattt gcaaagggaa
     1141 gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca tatttgagaa
     1201 gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata ctaaactcac
     1261 aaattagagc ttcaatttaa ttatatcagt tattacccta tgcggtgtga aataccgcac
     1321 agatgcgtaa ggagaaaata ccgcatcagg aaattgtaaa cgttaatatt ttgttaaaat
     1381 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     1441 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     1501 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     1561 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     1621 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     1681 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     1741 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     1801 gcgcgtccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     1861 ttcgctatta cgccagctga attggagcga cctcatgcta tacctgagaa agcaacctga
     1921 cctacaggaa agagttactc aagaataaga attttcgttt taaaacctaa gagtcacttt
     1981 aaaatttgta tacacttatt ttttttataa cttatttaat aataaaaatc ataaatcata
     2041 agaaattcgc ttatttagaa gtgtcaacaa cgtatctacc aacgatttga cccttttcca
     2101 tcttttcgta aatttctggc aaggtagaca agccgacaac cttgattgga gacttgacca
     2161 aacctctggc gaagaattgt taattaagag ctcagatctt atcgtcgtca tccttgtaat
     2221 ccatcgatac tagtgcggcc gccctttagt gagggttgaa ttcgaatttt caaaaattct
     2281 tacttttttt ttggatggac gcaaagaagt ttaataatca tattacatgg cattaccacc
     2341 atatacatat ccatatacat atccatatct aatcttactt atatgttgtg gaaatgtaaa
     2401 gagccccatt atcttagcct aaaaaaacct tctctttgga actttcagta atacgcttaa
     2461 ctgctcattg ctatattgaa gtacggatta gaagccgccg agcgggtgac agccctccga
     2521 aggaagactc tcctccgtgc gtcctcgtct tcaccggtcg cgttcctgaa acgcagatgt
     2581 gcctcgcgcc gcactgctcc gaacaataaa gattctacaa tactagcttt tatggttatg
     2641 aagaggaaaa attggcagta acctggcccc acaaaccttc aaatgaacga atcaaattaa
     2701 caaccatagg atgataatgc gattagtttt ttagccttat ttctggggta attaatcagc
     2761 gaagcgatga tttttgatct attaacagat atataaatgc aaaaactgca taaccacttt
     2821 aactaatact ttcaacattt tcggtttgta ttacttctta ttcaaatgta ataaaagtat
     2881 caacaaaaaa ttgttaatat acctctatac tttaacgtca aggagaaaaa accccggatc
     2941 cctcaatggc ttcttcagaa gagacctcgt ccattgatac caacgaactc atcacagatt
     3001 tgaaggaaaa gtgggatggt cttgagaaca aatctactgt acttatatat ggaggaggag
     3061 ccattgttgc tgtttggtta tcttccattg ttgttggtgc catcaactct gttcctctgc
     3121 ttccgaaagt tatggaactt gtcggactag ggtacactgg atggtttgtc tacagatacc
     3181 ttctcttcaa gtcaagcaga aaggaattgg ctgaggatat tgaatccttg aagaagaaga
     3241 tcgcaggaag cgaaggaggt ggtggatctg tgagcaaggg cgaggagctg ttcaccgggg
     3301 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     3361 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     3421 gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt cggctacggc ctgatgtgct
     3481 tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     3541 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     3601 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     3661 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     3721 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
     3781 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     3841 gccccgtgct gctgcccgac aaccactacc tgagctacca gtccaagctg agcaaagacc
     3901 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     3961 tcggcatgga cgagctgtac aagtaactcg agtaagcttg gtaccgcggc tagctaagat
     4021 ccgctctaac cgaaaaggaa ggagttagac aacctgaagt ctaggtccct atttattttt
     4081 ttatagttat gttagtatta agaacgttat ttatatttca aatttttctt ttttttctgt
     4141 acagacgcgt gtacgcatgt aacattatac tgaaaacctt gcttgagaag gttttgggac
     4201 gctcgaagat ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     4261 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4321 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4381 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4441 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4501 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4561 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4621 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4681 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4741 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4801 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4861 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     4921 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     4981 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5041 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5101 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     5161 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     5221 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     5281 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     5341 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     5401 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     5461 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     5521 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     5581 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     5641 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     5701 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     5761 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     5821 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     5881 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     5941 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     6001 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6061 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6121 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     6181 ttccccgaaa agtgccacct gaacgaagca tctgtgcttc attttgtaga acaaaaatgc
     6241 aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa
     6301 tgcaacgcga aagcgctatt ttaccaacga agaatctgtg cttcattttt gtaaaacaaa
     6361 aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac
     6421 agaaatgcaa cgcgagagcg ctattttacc aacaaagaat ctatacttct tttttgttct
     6481 acaaaaatgc atcccgagag cgctattttt ctaacaaagc atcttagatt actttttttc
     6541 tcctttgtgc gctctataat gcagtctctt gataactttt tgcactgtag gtccgttaag
     6601 gttagaagaa ggctactttg gtgtctattt tctcttccat aaaaaaagcc tgactccact
     6661 tcccgcgttt actgattact agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc
     6721 ccgattatat tctataccga tgtggattgc gcatactttg tgaacagaaa gtgatagcgt
     6781 tgatgattct tcattggtca gaaaattatg aacggtttct tctattttgt ctctatatac
     6841 tacgtatagg aaatgtttac attttcgtat tgttttcgat tcactctatg aatagttctt
     6901 actacaattt ttttgtctaa agagtaatac tagagataaa cataaaaaat gtagaggtcg
     6961 agtttagatg caagttcaag gagcgaaagg tggatgggta ggttatatag ggatatagca
     7021 cagagatata tagcaaagag atacttttga gcaatgtttg tggaagcggt attcgcaata
     7081 ttttagtagc tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg tcttcagagc
     7141 gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt ctagagaata ggaacttcgg
     7201 aataggaact tcaaagcgtt tccgaaaacg agcgcttccg aaaatgcaac gcgagctgcg
     7261 cacatacagc tcactgttca cgtcgcacct atatctgcgt gttgcctgta tatatatata
     7321 catgagaaga acggcatagt gcgtgtttat gcttaaatgc gtacttatat gcgtctattt
     7381 atgtaggatg aaaggtagtc tagtacctcc tgtgatatta tcccattcca tgcggggtat
     7441 cgtatgcttc cttcagcact accctttagc tgttctatat gctgccactc ctcaattgga
     7501 ttagtctcat ccttcaatgc tatcatttcc tttgatattg gatcatacta agaaaccatt
     7561 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tc
//
