LOCUS       sequence-352568-        6794 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses yeGFP under the control of a synthetic promoter (PGAL7-A).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6794)
  AUTHORS   Kwak S, Mahmud B, Dantas G
  TITLE     A Tunable and Expandable Transactivation System in Probiotic Yeast 
            Saccharomyces boulardii.
  JOURNAL   ACS Synth Biol. 2022 Jan 21;11(1):508-514. doi: 
            10.1021/acssynbio.1c00384. Epub 2021 Dec 23.
  PUBMED    34939781
REFERENCE   2  (bases 1 to 6794)
  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.1c00384"; journalName: "ACS Synth Biol"; date: 
            "2022-01-21- 21"; volume: "11"; issue: "1"; pages: "508-514"
COMMENT     Sequence Label: pRS426-PGAL7-A_GFP_TCYC1
FEATURES             Location/Qualifiers
     source          1..6794
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     14..36
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(74..92)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      133..1475
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    465..512
                     /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)."
     promoter        1502..1606
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1607..1675,1676..2467)
                     /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     complement(1825..1844)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2638..3226
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3127..3146
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3380..3397
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3514..3535
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3550..3555,3556..3573,3574..3580)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3588..3604
                     /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)."
     primer_bind     3593..3615
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3612..3628
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3612..3628
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3647..3667
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3649..3667
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             3886..4602
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             3886..4599
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      4609..4856
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4625..4643)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4874..4893)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4875..4893)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4903..4920)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4903..4919)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4912..4934)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5060..5515
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5147..5166)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5357..5378
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5646..6449)
                     /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"
     promoter        complement(6450..6665)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(6689..6708)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
       61 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
      121 gtttcttagt atgatccaat atcaaaggaa atgatagcat tgaaggatga gactaatcca
      181 attgaggagt ggcagcatat agaacagcta aagggtagtg ctgaaggaag catacgatac
      241 cccgcatgga atgggataat atcacaggag gtactagact acctttcatc ctacataaat
      301 agacgcatat aagtacgcat ttaagcataa acacgcacta tgccgttctt ctcatgtata
      361 tatatataca ggcaacacgc agatataggt gcgacgtgaa cagtgagctg tatgtgcgca
      421 gctcgcgttg cattttcgga agcgctcgtt ttcggaaacg ctttgaagtt cctattccga
      481 agttcctatt ctctagaaag tataggaact tcagagcgct tttgaaaacc aaaagcgctc
      541 tgaagacgca ctttcaaaaa accaaaaacg caccggactg taacgagcta ctaaaatatt
      601 gcgaataccg cttccacaaa cattgctcaa aagtatctct ttgctatata tctctgtgct
      661 atatccctat ataacctacc catccacctt tcgctccttg aacttgcatc taaactcgac
      721 ctctacattt tttatgttta tctctagtat tactctttag acaaaaaaat tgtagtaaga
      781 actattcata gagtgaatcg aaaacaatac gaaaatgtaa acatttccta tacgtagtat
      841 atagagacaa aatagaagaa accgttcata attttctgac caatgaagaa tcatcaacgc
      901 tatcactttc tgttcacaaa gtatgcgcaa tccacatcgg tatagaatat aatcggggat
      961 gcctttatct tgaaaaaatg cacccgcagc ttcgctagta atcagtaaac gcgggaagtg
     1021 gagtcaggct ttttttatgg aagagaaaat agacaccaaa gtagccttct tctaacctta
     1081 acggacctac agtgcaaaaa gttatcaaga gactgcatta tagagcgcac aaaggagaaa
     1141 aaaagtaatc taagatgctt tgttagaaaa atagcgctct cgggatgcat ttttgtagaa
     1201 caaaaaagaa gtatagattc tttgttggta aaatagcgct ctcgcgttgc atttctgttc
     1261 tgtaaaaatg cagctcagat tctttgtttg aaaaattagc gctctcgcgt tgcatttttg
     1321 ttttacaaaa atgaagcaca gattcttcgt tggtaaaata gcgctttcgc gttgcatttc
     1381 tgttctgtaa aaatgcagct cagattcttt gtttgaaaaa ttagcgctct cgcgttgcat
     1441 ttttgttcta caaaatgaag cacagatgct tcgttcaggt ggcacttttc ggggaaatgt
     1501 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     1561 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     1621 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     1681 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     1741 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     1801 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     1861 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     1921 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     1981 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     2041 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     2101 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     2161 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     2221 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     2281 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     2341 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     2401 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     2461 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     2521 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     2581 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     2641 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     2701 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     2761 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     2821 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     2881 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     2941 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     3001 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     3061 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     3121 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     3181 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     3241 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     3301 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     3361 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     3421 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     3481 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
     3541 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     3601 acaatttcac acaggaaaca gctatgacca tgattacgcc aagcgcgcaa ttaaccctca
     3661 ctaaagggaa caaaagctgg agctcatctg tcgacagtgc gatacaggac tggctataca
     3721 gtgttcacaa aatagccaag ctgaaaataa tgtgtagcta tgttcagtta gtttggctag
     3781 caaagatata aaagcaggtc ggaaatattt atgggcatta ttatgcagag catcaacatg
     3841 ataaaaaaaa acagttgaat attccctcaa aaaactagta aaacaatgtc taaaggtgaa
     3901 gaattattca ctggtgttgt cccaattttg gttgaattag atggtgatgt taatggtcac
     3961 aaattttctg tctccggtga aggtgaaggt gatgctactt acggtaaatt gaccttaaaa
     4021 tttatttgta ctactggtaa attgccagtt ccatggccaa ccttagtcac tactttaact
     4081 tatggtgttc aatgtttttc tagataccca gatcatatga aacaacatga ctttttcaag
     4141 tctgccatgc cagaaggtta tgttcaagaa agaactattt ttttcaaaga tgacggtaac
     4201 tacaagacca gagctgaagt caagtttgaa ggtgatacct tagttaatag aatcgaatta
     4261 aaaggtattg attttaaaga agatggtaac attttaggtc acaaattgga atacaactat
     4321 aactctcaca atgtttacat catggctgac aaacaaaaga atggtatcaa agttaacttc
     4381 aaaattagac acaacattga agatggttct gttcaattag ctgaccatta tcaacaaaat
     4441 actccaattg gtgatggtcc agtcttgtta ccagacaacc attacttatc cactcaatct
     4501 gccttatcca aagatccaaa cgaaaagaga gaccacatgg tcttgttaga atttgttact
     4561 gctgctggta ttacccatgg tatggatgaa ttgtacaaat aactcgagtc atgtaattag
     4621 ttatgtcacg cttacattca cgccctcccc ccacatccgc tctaaccgaa aaggaaggag
     4681 ttagacaacc tgaagtctag gtccctattt atttttttat agttatgtta gtattaagaa
     4741 cgttatttat atttcaaatt tttctttttt ttctgtacag acgcgtgtac gcatgtaaca
     4801 ttatactgaa aaccttgctt gagaaggttt tgggacgctc gaaggcttta atttgcggcc
     4861 ggtacccaat tcgccctata gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca
     4921 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
     4981 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     5041 cagcctgaat ggcgaatgga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     5101 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     5161 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     5221 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     5281 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     5341 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     5401 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     5461 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttcctga
     5521 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcataggg taataactga
     5581 tataattaaa ttgaagctct aatttgtgag tttagtatac atgcatttac ttataataca
     5641 gttttttagt tttgctggcc gcatcttctc aaatatgctt cccagcctgc ttttctgtaa
     5701 cgttcaccct ctaccttagc atcccttccc tttgcaaata gtcctcttcc aacaataata
     5761 atgtcagatc ctgtagagac cacatcatcc acggttctat actgttgacc caatgcgtct
     5821 cccttgtcat ctaaacccac accgggtgtc ataatcaacc aatcgtaacc ttcatctctt
     5881 ccacccatgt ctctttgagc aataaagccg ataacaaaat ctttgtcgct cttcgcaatg
     5941 tcaacagtac ccttagtata ttctccagta gatagggagc ccttgcatga caattctgct
     6001 aacatcaaaa ggcctctagg ttcctttgtt acttcttctg ccgcctgctt caaaccgcta
     6061 acaatacctg ggcccaccac accgtgtgca ttcgtaatgt ctgcccattc tgctattctg
     6121 tatacacccg cagagtactg caatttgact gtattaccaa tgtcagcaaa ttttctgtct
     6181 tcgaagagta aaaaattgta cttggcggat aatgccttta gcggcttaac tgtgccctcc
     6241 atggaaaaat cagtcaagat atccacatgt gtttttagta aacaaatttt gggacctaat
     6301 gcttcaacta actccagtaa ttccttggtg gtacgaacat ccaatgaagc acacaagttt
     6361 gtttgctttt cgtgcatgat attaaatagc ttggcagcaa caggactagg atgagtagca
     6421 gcacgttcct tatatgtagc tttcgacatg atttatcttc gtttcctgca ggtttttgtt
     6481 ctgtgcagtt gggttaagaa tactgggcaa tttcatgttt cttcaacact acatatgcgt
     6541 atatatacca atctaagtct gtgctccttc cttcgttctt ccttctgttc ggagattacc
     6601 gaatcaaaaa aatttcaagg aaaccgaaat caaaaaaaag aataaaaaaa aaatgatgaa
     6661 ttgaaaaggt ggtatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     6721 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     6781 ccgcttacag acaa
//
