LOCUS       sequence-352585-        6788 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses yeRFP under the control of a synthetic promoter (PGAL7-D).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6788)
  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 6788)
  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-D_RFP_TCYC1
FEATURES             Location/Qualifiers
     source          1..6788
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     15..37
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(75..93)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      134..1476
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    466..513
                     /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        1503..1607
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1608..1676,1677..2468)
                     /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(1826..1845)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2639..3227
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3128..3147
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3381..3398
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3515..3536
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3551..3556,3557..3574,3575..3581)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3589..3605
                     /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     3594..3616
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3613..3629
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3613..3629
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3648..3668
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3650..3668
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    3687..3705
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      4604..4851
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4620..4638)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4869..4888)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4870..4888)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4898..4915)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4898..4914)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4907..4929)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5055..5510
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5142..5161)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5352..5373
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5641..6444)
                     /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(6445..6660)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(6684..6703)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
       61 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
      121 ggtttcttag tatgatccaa tatcaaagga aatgatagca ttgaaggatg agactaatcc
      181 aattgaggag tggcagcata tagaacagct aaagggtagt gctgaaggaa gcatacgata
      241 ccccgcatgg aatgggataa tatcacagga ggtactagac tacctttcat cctacataaa
      301 tagacgcata taagtacgca tttaagcata aacacgcact atgccgttct tctcatgtat
      361 atatatatac aggcaacacg cagatatagg tgcgacgtga acagtgagct gtatgtgcgc
      421 agctcgcgtt gcattttcgg aagcgctcgt tttcggaaac gctttgaagt tcctattccg
      481 aagttcctat tctctagaaa gtataggaac ttcagagcgc ttttgaaaac caaaagcgct
      541 ctgaagacgc actttcaaaa aaccaaaaac gcaccggact gtaacgagct actaaaatat
      601 tgcgaatacc gcttccacaa acattgctca aaagtatctc tttgctatat atctctgtgc
      661 tatatcccta tataacctac ccatccacct ttcgctcctt gaacttgcat ctaaactcga
      721 cctctacatt ttttatgttt atctctagta ttactcttta gacaaaaaaa ttgtagtaag
      781 aactattcat agagtgaatc gaaaacaata cgaaaatgta aacatttcct atacgtagta
      841 tatagagaca aaatagaaga aaccgttcat aattttctga ccaatgaaga atcatcaacg
      901 ctatcacttt ctgttcacaa agtatgcgca atccacatcg gtatagaata taatcgggga
      961 tgcctttatc ttgaaaaaat gcacccgcag cttcgctagt aatcagtaaa cgcgggaagt
     1021 ggagtcaggc tttttttatg gaagagaaaa tagacaccaa agtagccttc ttctaacctt
     1081 aacggaccta cagtgcaaaa agttatcaag agactgcatt atagagcgca caaaggagaa
     1141 aaaaagtaat ctaagatgct ttgttagaaa aatagcgctc tcgggatgca tttttgtaga
     1201 acaaaaaaga agtatagatt ctttgttggt aaaatagcgc tctcgcgttg catttctgtt
     1261 ctgtaaaaat gcagctcaga ttctttgttt gaaaaattag cgctctcgcg ttgcattttt
     1321 gttttacaaa aatgaagcac agattcttcg ttggtaaaat agcgctttcg cgttgcattt
     1381 ctgttctgta aaaatgcagc tcagattctt tgtttgaaaa attagcgctc tcgcgttgca
     1441 tttttgttct acaaaatgaa gcacagatgc ttcgttcagg tggcactttt cggggaaatg
     1501 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     1561 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     1621 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     1681 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     1741 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     1801 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     1861 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     1921 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     1981 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     2041 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     2101 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     2161 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     2221 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     2281 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     2341 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     2401 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     2461 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     2521 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     2581 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     2641 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     2701 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     2761 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     2821 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     2881 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     2941 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     3001 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     3061 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     3121 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     3181 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     3241 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     3301 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     3361 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     3421 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     3481 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
     3541 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
     3601 aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc
     3661 actaaaggga acaaaagctg gagctctctc tatcactgat agggagcgga ctggctatac
     3721 agtgttaaca aaatagccaa gctgaaaata atgtgtagct atgttcagtt agtttggcta
     3781 gcaaagatat aaaagcaggt cggaaatatt tatgggcatt attatgcaga gcatcaacat
     3841 gataaaaaaa aacagttgaa tattccctca aaaaactagt aaaacaatgg taagtaaggg
     3901 tgaagaagac aatatggcga tcattaagga attcatgcgt ttcaaagtac acatggaggg
     3961 aagcgtgaac ggacatgaat ttgaaatcga aggggaaggc gaaggtagac catacgaagg
     4021 aacccagacc gcaaagctta aagttaccaa aggcgggcca ctaccatttg catgggatat
     4081 cttgagccct cagtttatgt atggcagtaa ggcctacgtt aaacacccag ctgatattcc
     4141 cgactatttg aaattgtctt ttccagaagg attcaaatgg gaaagagtaa tgaatttcga
     4201 ggacggcgga gttgttactg ttactcaaga ttcaagtttg caagacggtg aatttattta
     4261 caaggtcaaa ttaagaggga ctaatttccc tagtgatggt cccgtcatgc aaaagaagac
     4321 tatgggttgg gaagcctcat ctgaacgtat gtatccagaa gatggcgcgc ttaaggggga
     4381 aattaaacaa agattgaagt taaaagacgg tggtcactac gacgcggaag ttaagaccac
     4441 ttataaagct aaaaagcccg ttcagttacc tggtgcatat aacgtaaaca ttaaattgga
     4501 tatcacttca cataatgaag attacactat tgtggaacaa tatgaaagag ctgaaggtag
     4561 gcactcaacg ggtggaatgg acgaattgta caaataactc gagtcatgta attagttatg
     4621 tcacgcttac attcacgccc tccccccaca tccgctctaa ccgaaaagga aggagttaga
     4681 caacctgaag tctaggtccc tatttatttt tttatagtta tgttagtatt aagaacgtta
     4741 tttatatttc aaatttttct tttttttctg tacagacgcg tgtacgcatg taacattata
     4801 ctgaaaacct tgcttgagaa ggttttggga cgctcgaagg ctttaatttg cggccggtac
     4861 ccaattcgcc ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc
     4921 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     4981 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     5041 tgaatggcga atggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     5101 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     5161 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     5221 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     5281 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     5341 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     5401 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     5461 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt cctgatgcgg
     5521 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tagggtaata actgatataa
     5581 ttaaattgaa gctctaattt gtgagtttag tatacatgca tttacttata atacagtttt
     5641 ttagttttgc tggccgcatc ttctcaaata tgcttcccag cctgcttttc tgtaacgttc
     5701 accctctacc ttagcatccc ttccctttgc aaatagtcct cttccaacaa taataatgtc
     5761 agatcctgta gagaccacat catccacggt tctatactgt tgacccaatg cgtctccctt
     5821 gtcatctaaa cccacaccgg gtgtcataat caaccaatcg taaccttcat ctcttccacc
     5881 catgtctctt tgagcaataa agccgataac aaaatctttg tcgctcttcg caatgtcaac
     5941 agtaccctta gtatattctc cagtagatag ggagcccttg catgacaatt ctgctaacat
     6001 caaaaggcct ctaggttcct ttgttacttc ttctgccgcc tgcttcaaac cgctaacaat
     6061 acctgggccc accacaccgt gtgcattcgt aatgtctgcc cattctgcta ttctgtatac
     6121 acccgcagag tactgcaatt tgactgtatt accaatgtca gcaaattttc tgtcttcgaa
     6181 gagtaaaaaa ttgtacttgg cggataatgc ctttagcggc ttaactgtgc cctccatgga
     6241 aaaatcagtc aagatatcca catgtgtttt tagtaaacaa attttgggac ctaatgcttc
     6301 aactaactcc agtaattcct tggtggtacg aacatccaat gaagcacaca agtttgtttg
     6361 cttttcgtgc atgatattaa atagcttggc agcaacagga ctaggatgag tagcagcacg
     6421 ttccttatat gtagctttcg acatgattta tcttcgtttc ctgcaggttt ttgttctgtg
     6481 cagttgggtt aagaatactg ggcaatttca tgtttcttca acactacata tgcgtatata
     6541 taccaatcta agtctgtgct ccttccttcg ttcttccttc tgttcggaga ttaccgaatc
     6601 aaaaaaattt caaggaaacc gaaatcaaaa aaaagaataa aaaaaaaatg atgaattgaa
     6661 aaggtggtat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     6721 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     6781 tacagaca
//
