LOCUS       sequence-229428-        7284 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mCitrine controlled by the Gal1 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7284)
  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 7284)
  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/mCitrine
FEATURES             Location/Qualifiers
     source          1..7284
                     /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(679..681,682..684,685..1398)
                     /codon_start=1
                     /product="enhanced YFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEMFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(724..745)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(985..1004)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1332..1353
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      1431..1620
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1692..1709)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1863..2451)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1943..1962)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2622..3413,3414..3482))
                     /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     3245..3264
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3483..3587)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3614..4956
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(4577..4624)
                     /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     4997..5015
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5053..5075)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5175..5194
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5218..5433
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             5434..6237
                     /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(6368..6823)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6505..6526)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6717..6736
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(6902..7067)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     6988..7012
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(7215..7238)
                     /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 ggtgagcaag ggcgaggaga tgttcaccgg ggtggtgccc
      721 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
      781 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
      841 cccgtgccct ggcccaccct cgtgaccacc ttcggctacg gcctgatgtg cttcgcccgc
      901 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
      961 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     1021 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     1081 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
     1141 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
     1201 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     1261 ctgctgcccg acaaccacta cctgagctac cagtccaagc tgagcaaaga ccccaacgag
     1321 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     1381 gacgagctgt acaagtagct cgagtaagct tggtaccgcg gctagctaag atccgctcta
     1441 accgaaaagg aaggagttag acaacctgaa gtctaggtcc ctatttattt ttttatagtt
     1501 atgttagtat taagaacgtt atttatattt caaatttttc ttttttttct gtacagacgc
     1561 gtgtacgcat gtaacattat actgaaaacc ttgcttgaga aggttttggg acgctcgaag
     1621 atccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     1681 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     1741 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     1801 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     1861 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     1921 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     1981 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     2041 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     2101 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     2161 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     2221 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     2281 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     2341 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     2401 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     2461 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     2521 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     2581 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     2641 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     2701 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     2761 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     2821 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     2881 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     2941 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     3001 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     3061 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     3121 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     3181 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     3241 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     3301 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     3361 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     3421 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     3481 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     3541 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     3601 aaagtgccac ctgaacgaag catctgtgct tcattttgta gaacaaaaat gcaacgcgag
     3661 agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc
     3721 gaaagcgcta ttttaccaac gaagaatctg tgcttcattt ttgtaaaaca aaaatgcaac
     3781 gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc
     3841 aacgcgagag cgctatttta ccaacaaaga atctatactt cttttttgtt ctacaaaaat
     3901 gcatcccgag agcgctattt ttctaacaaa gcatcttaga ttactttttt tctcctttgt
     3961 gcgctctata atgcagtctc ttgataactt tttgcactgt aggtccgtta aggttagaag
     4021 aaggctactt tggtgtctat tttctcttcc ataaaaaaag cctgactcca cttcccgcgt
     4081 ttactgatta ctagcgaagc tgcgggtgca ttttttcaag ataaaggcat ccccgattat
     4141 attctatacc gatgtggatt gcgcatactt tgtgaacaga aagtgatagc gttgatgatt
     4201 cttcattggt cagaaaatta tgaacggttt cttctatttt gtctctatat actacgtata
     4261 ggaaatgttt acattttcgt attgttttcg attcactcta tgaatagttc ttactacaat
     4321 ttttttgtct aaagagtaat actagagata aacataaaaa atgtagaggt cgagtttaga
     4381 tgcaagttca aggagcgaaa ggtggatggg taggttatat agggatatag cacagagata
     4441 tatagcaaag agatactttt gagcaatgtt tgtggaagcg gtattcgcaa tattttagta
     4501 gctcgttaca gtccggtgcg tttttggttt tttgaaagtg cgtcttcaga gcgcttttgg
     4561 ttttcaaaag cgctctgaag ttcctatact ttctagagaa taggaacttc ggaataggaa
     4621 cttcaaagcg tttccgaaaa cgagcgcttc cgaaaatgca acgcgagctg cgcacataca
     4681 gctcactgtt cacgtcgcac ctatatctgc gtgttgcctg tatatatata tacatgagaa
     4741 gaacggcata gtgcgtgttt atgcttaaat gcgtacttat atgcgtctat ttatgtagga
     4801 tgaaaggtag tctagtacct cctgtgatat tatcccattc catgcggggt atcgtatgct
     4861 tccttcagca ctacccttta gctgttctat atgctgccac tcctcaattg gattagtctc
     4921 atccttcaat gctatcattt cctttgatat tggatcatac taagaaacca ttattatcat
     4981 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     5041 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     5101 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     5161 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata ccaccttttc
     5221 aattcatcat ttttttttta ttcttttttt tgatttcggt ttccttgaaa tttttttgat
     5281 tcggtaatct ccgaacagaa ggaagaacga aggaaggagc acagacttag attggtatat
     5341 atacgcatat gtagtgttga agaaacatga aattgcccag tattcttaac ccaactgcac
     5401 agaacaaaaa cctgcaggaa acgaagataa atcatgtcga aagctacata taaggaacgt
     5461 gctgctactc atcctagtcc tgttgctgcc aagctattta atatcatgca cgaaaagcaa
     5521 acaaacttgt gtgcttcatt ggatgttcgt accaccaagg aattactgga gttagttgaa
     5581 gcattaggtc ccaaaatttg tttactaaaa acacatgtgg atatcttgac tgatttttcc
     5641 atggagggca cagttaagcc gctaaaggca ttatccgcca agtacaattt tttactcttc
     5701 gaagacagaa aatttgctga cattggtaat acagtcaaat tgcagtactc tgcgggtgta
     5761 tacagaatag cagaatgggc agacattacg aatgcacacg gtgtggtggg cccaggtatt
     5821 gttagcggtt tgaagcaggc ggcagaagaa gtaacaaagg aacctagagg ccttttgatg
     5881 ttagcagaat tgtcatgcaa gggctcccta tctactggag aatatactaa gggtactgtt
     5941 gacattgcga agagcgacaa agattttgtt atcggcttta ttgctcaaag agacatgggt
     6001 ggaagagatg aaggttacga ttggttgatt atgacacccg gtgtgggttt agatgacaag
     6061 ggagacgcat tgggtcaaca gtatagaacc gtggatgatg tggtctctac aggatctgac
     6121 attattattg ttggaagagg actatttgca aagggaaggg atgctaaggt agagggtgaa
     6181 cgttacagaa aagcaggctg ggaagcatat ttgagaagat gcggccagca aaactaaaaa
     6241 actgtattat aagtaaatgc atgtatacta aactcacaaa ttagagcttc aatttaatta
     6301 tatcagttat taccctatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     6361 catcaggaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     6421 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     6481 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     6541 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     6601 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     6661 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     6721 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     6781 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtccattcg ccattcaggc
     6841 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctgaatt
     6901 ggagcgacct catgctatac ctgagaaagc aacctgacct acaggaaaga gttactcaag
     6961 aataagaatt ttcgttttaa aacctaagag tcactttaaa atttgtatac acttattttt
     7021 tttataactt atttaataat aaaaatcata aatcataaga aattcgctta tttagaagtg
     7081 tcaacaacgt atctaccaac gatttgaccc ttttccatct tttcgtaaat ttctggcaag
     7141 gtagacaagc cgacaacctt gattggagac ttgaccaaac ctctggcgaa gaattgttaa
     7201 ttaagagctc agatcttatc gtcgtcatcc ttgtaatcca tcgatactag tgcggccgcc
     7261 ctttagtgag ggttgaattc gaat
//
