LOCUS       sequence-417211-        8976 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses CYP tagged with ymsfGFP in S. cerevisiae.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8976)
  AUTHORS   Chadwick SR, Stack-Couture S, Berg MD, Gregorio SD, Lung B, 
            Genereaux J, Moir RD, Brandl CJ, Willis IM, Snapp EL, Lajoie P
  TITLE     TUDCA modulates drug bioavailability to regulate resistance to acute
            ER stress in Saccharomyces cerevisiae.
  JOURNAL   Mol Biol Cell. 2024 Dec 11:mbcE24040147. doi: 
            10.1091/mbc.E24-04-0147.
  PUBMED    39661468
REFERENCE   2  (bases 1 to 8976)
  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: "Mol Biol 
            Cell. 2024 Dec 11:mbcE24040147. doi: 10.1091/mbc.E24-04-0147."
COMMENT     Sequence Label: CPY-ymsfGFP
FEATURES             Location/Qualifiers
     source          1..8976
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(203..222)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     322..344
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(382..400)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    442..945
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        982..1086
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1087..1155,1156..1947)
                     /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(1305..1324)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2118..2706
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2607..2626
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2860..2877
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2994..3015
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3030..3035,3036..3053,3054..3060)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3068..3084
                     /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     3073..3095
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3092..3108
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3092..3108
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3127..3147
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3129..3147
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3171..3612
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    3171..3288
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     3583..3606
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     terminator      5976..6223
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5992..6010)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(6241..6260)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6242..6260)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6270..6287)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6270..6286)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6279..6301)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6427..6882
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6514..6533)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6724..6745
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7183..7590
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             7591..8685
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
ORIGIN
        1 atgattccta atttatatat tggaggattt tctctaaaaa aaaaaaaata caacaaataa
       61 aaaacactca atgacctgac catttgatgg agtttaagtc aataccttct tgaaccattt
      121 cccataatgg tgaaagttcc ctcaagaatt ttactctgtc agaaacggcc ttaacgacgt
      181 agtcgatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
      241 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
      301 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
      361 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
      421 taataatggt ttcttaggac ggatcgcttg cctgtaactt acacgcgcct cgtatctttt
      481 aatgatggaa taatttggga atttactctg tgtttattta tttttatgtt ttgtatttgg
      541 attttagaaa gtaaataaag aaggtagaag agttacggaa tgaagaaaaa aaaataaaca
      601 aaggtttaaa aaatttcaac aaaaagcgta ctttacatat atatttatta gacaagaaaa
      661 gcagattaaa tagatataca ttcgattaac gataagtaaa atgtaaaatc acaggatttt
      721 cgtgtgtggt cttctacaca gacaagatga aacaattcgg cattaatacc tgagagcagg
      781 aagagcaaga taaaaggtag tatttgttgg cgatccccct agagtctttt acatcttcgg
      841 aaaacaaaaa ctattttttc tttaatttct ttttttactt tctattttta atttatatat
      901 ttatattaaa aaatttaaat tataattatt tttatagcac gtgatgaaaa ggacccaggt
      961 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     1021 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     1081 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     1141 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     1201 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     1261 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     1321 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     1381 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     1441 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     1501 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     1561 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     1621 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     1681 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     1741 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     1801 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     1861 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     1921 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     1981 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     2041 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     2101 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     2161 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     2221 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     2281 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     2341 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     2401 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     2461 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     2521 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     2581 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     2641 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     2701 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     2761 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     2821 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     2881 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     2941 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     3001 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     3061 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
     3121 aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg agctctagta cggattagaa
     3181 gccgccgagc gggtgacagc cctccgaagg aagactctcc tccgtgcgtc ctcgtcttca
     3241 ccggtcgcgt tcctgaaacg cagatgtgcc tcgcgccgca ctgctccgaa caataaagat
     3301 tctacaatac tagcttttat ggttatgaag aggaaaaatt ggcagtaacc tggccccaca
     3361 aaccttcaaa tgaacgaatc aaattaacaa ccataggatg ataatgcgat tagtttttta
     3421 gccttatttc tggggtaatt aatcagcgaa gcgatgattt ttgatctatt aacagatata
     3481 taaatgcaaa aactgcataa ccactttaac taatactttc aacattttcg gtttgtatta
     3541 cttcttattc aaatgtaata aaagtatcaa caaaaaattg ttaatatacc tctatacttt
     3601 aacgtcaagg agaaaaaacc ccggattcta gaactagtct aaaaatgaaa gcattcacca
     3661 gtttactatg tggactaggc ctgtccacta cactcgctaa ggccatctca ttgcaaagac
     3721 cgttgggtct agataaggac gttttgctgc aagctgcgga aaaatttggt ttggacctcg
     3781 acctggatca tctcttgaag gagttggact ccaatgtatt ggacgcttgg gcccaaatag
     3841 agcatttgta cccaaaccag gttatgagcc ttgaaacttc cactaagcca aaattccctg
     3901 aagcaatcaa aacgaagaaa gactgggact ttgtggtcaa gaatgacgca attgaaaact
     3961 atcagcttcg tgtcaacaag attaaggacc ctaaaatcct gggcattgac ccaaatgtca
     4021 cacagtacac gggttacttg gatgtggaag acgaggacaa gcatttcttc ttttggactt
     4081 ttgaaagtag aaacgatcct gcaaaggatc cggtcatcct ttggttgaac gggggtccag
     4141 gttgttcttc actaaccggg ctgttctttg aattaggacc ctcatccatt ggacctgatt
     4201 tgaaacccat cgggaaccct tactcttgga acagcaatgc caccgtgatc ttccttgacc
     4261 agcctgtcaa cgttgggttc tcgtattccg ggtcctcagg tgtttccaac actgtcgccg
     4321 ctggtaagga tgtctataac ttcttggagt tgttcttcga tcagttccct gaatacgtca
     4381 acaagggcca agatttccac atcgctgggg aatcctacgc cggccattac atccctgttt
     4441 ttgcctctga aattttgtct cacaaggaca gaaacttcaa cttaacctcc gtcttgatcg
     4501 gaaatggcct cactgaccca ttgactcagt ataactatta cgaaccaatg gcctgtggtg
     4561 aaggtggcga accatctgtt ttgccctcgg aggaatgctc tgctatggaa gactctttgg
     4621 aacgttgttt gggcttgatc gagtcgtgct atgactcgca atccgtctgg tcctgtgttc
     4681 cagctaccat ttattgtaat aacgcccaat tggctcctta ccaacgtacc ggcagaaacg
     4741 tttacgatat caggaaggat tgtgaaggtg gcaatttgtg ctacccaacg ttacaagata
     4801 tcgacgacta cttaaaccag gactacgtca aagaagctgt cggtgcggag gttgaccact
     4861 acgaatcctg taacttcgat atcaacagaa atttcctgtt tgcgggtgat tggatgaagc
     4921 cttaccacac cgccgtaaca gatcttttga atcaagacct acccattctg gtatatgcag
     4981 gcgataaaga tttcatctgt aactggttgg gtaataaggc gtggacggat gtcttgccat
     5041 ggaagtacga cgaagaattt gcaagccaaa aagtacgtaa ctggactgct tctatcaccg
     5101 acgaggtcgc tggtgaagtc aaatcctaca agcacttcac ctatttgaga gtcttcaatg
     5161 gtggccacat ggttccattt gacgtccctg aaaacgcctt aagtatggtt aacgaatgga
     5221 tccacggtgg tttctcctta aagcttgtat ccaaaggtga agaattcttt actggtgttg
     5281 ttcctatttt agttgaatta gacggtgacg ttaatggtca taagtttagt gttagaggtg
     5341 aaggtgaagg tgacgcaact aacggtaaat tgacattgaa gtttatttgt actactggta
     5401 aattgccagt tccatggcca accttggtta ccactttaac ttatggtgtt caatgcttct
     5461 caagataccc tgatcatatg aaaagacacg actttttcaa gagtgccatg ccagaaggtt
     5521 acgtacaaga aagaactata tcttttaaag atgacggtac atacaagacc agagctgaag
     5581 taaaattcga aggtgacaca ttggtcaaca gaatcgaatt gaagggtatc gatttcaagg
     5641 aagacggtaa catcttgggt cataaattag aatacaattt caactcacac aatgtttaca
     5701 tcactgctga taagcaaaag aatggtatca aagcaaactt caagatcaga cataacgttg
     5761 aagatggttc cgtacaatta gctgaccact atcaacaaaa tacacctatt ggtgacggtc
     5821 ctgttttgtt accagacaac cattacttgt ctacccaatc aaaattgtcc aaagatccaa
     5881 acgaaaagag agaccatatg gtcttgttgg aatttgtcac tgccgctggt atcactcacg
     5941 gtatggatga attgtataag taggtcgacc tcgagtcatg taattagtta tgtcacgctt
     6001 acattcacgc cctcccccca catccgctct aaccgaaaag gaaggagtta gacaacctga
     6061 agtctaggtc cctatttatt tttttatagt tatgttagta ttaagaacgt tatttatatt
     6121 tcaaattttt cttttttttc tgtacagacg cgtgtacgca tgtaacatta tactgaaaac
     6181 cttgcttgag aaggttttgg gacgctcgaa ggctttaatt tgcggccggt acccaattcg
     6241 ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg
     6301 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     6361 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     6421 gaatggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     6481 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     6541 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     6601 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     6661 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     6721 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     6781 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     6841 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttcctgatgc ggtattttct
     6901 ccttacgcat ctgtgcggta tttcacaccg catatcgacc ggtcgaggag aacttctagt
     6961 atatctacat acctaatatt attgccttat taaaaatgga atcccaacaa ttacatcaaa
     7021 atccacattc tcttcaaaat caattgtcct gtacttcctt gttcatgtgt gttcaaaaac
     7081 gttatattta taggataatt atactctatt tctcaacaag taattggttg tttggccgag
     7141 cggtctaagg cgcctgattc aagaaatatc ttgaccgcag ttaactgtgg gaatactcag
     7201 gtatcgtaag atgcaagagt tcgaatctct tagcaaccat tatttttttc ctcaacataa
     7261 cgagaacaca caggggcgct atcgcacaga atcaaattcg atgactggaa attttttgtt
     7321 aatttcagag gtcgcctgac gcatatacct ttttcaactg aaaaattggg agaaaaagga
     7381 aaggtgagag cgccggaacc ggcttttcat atagaataga gaagcgttca tgactaaatg
     7441 cttgcatcac aatacttgaa gttgacaata ttatttaagg acctattgtt ttttccaata
     7501 ggtggttagc aatcgtctta ctttctaact tttcttacct tttacatttc agcaatatat
     7561 atatatatat ttcaaggata taccattcta atgtctgccc ctaagaagat cgtcgttttg
     7621 ccaggtgacc acgttggtca agaaatcaca gccgaagcca ttaaggttct taaagctatt
     7681 tctgatgttc gttccaatgt caagttcgat ttcgaaaatc atttaattgg tggtgctgct
     7741 atcgatgcta caggtgttcc acttccagat gaggcgctgg aagcctccaa gaaggctgat
     7801 gccgttttgt taggtgctgt gggtggtcct aaatggggta ccggtagtgt tagacctgaa
     7861 caaggtttac taaaaatccg taaagaactt caattgtacg ccaacttaag accatgtaac
     7921 tttgcatccg actctctttt agacttatct ccaatcaagc cacaatttgc taaaggtact
     7981 gacttcgttg ttgtcagaga attagtggga ggtatttact ttggtaagag aaaggaagac
     8041 gatggtgatg gtgtcgcttg ggatagtgaa caatacaccg ttccagaagt gcaaagaatc
     8101 acaagaatgg ccgctttcat ggccctacaa catgagccac cattgcctat ttggtccttg
     8161 gataaagcta atgttttggc ctcttcaaga ttatggagaa aaactgtgga ggaaaccatc
     8221 aagaacgaat tccctacatt gaaggttcaa catcaattga ttgattctgc cgccatgatc
     8281 ctagttaaga acccaaccca cctaaatggt attataatca ccagcaacat gtttggtgat
     8341 atcatctccg atgaagcctc cgttatccca ggttccttgg gtttgttgcc atctgcgtcc
     8401 ttggcctctt tgccagacaa gaacaccgca tttggtttgt acgaaccatg ccacggttct
     8461 gctccagatt tgccaaagaa taaggtcaac cctatcgcca ctatcttgtc tgctgcaatg
     8521 atgttgaaat tgtcattgaa cttgcctgaa gaaggtaagg ccattgaaga tgcagttaaa
     8581 aaggttttgg atgcaggtat cagaactggt gatttaggtg gttccaacag taccaccgaa
     8641 gtcggtgatg ctgtcgccga agaagttaag aaaatccttg cttaaaaaga ttctcttttt
     8701 ttatgatatt tgtacataaa ctttataaat gaaattcata atagaaacga cacgaaatta
     8761 caaaatggaa tatgttcata gggtagacga aactatatac gcaatctaca tacatttatc
     8821 aagaaggaga aaaaggagga tgtaaaggaa tacaggtaag caaattgata ctaatggctc
     8881 aacgtgataa ggaaaaagaa ttgcacttta acattaatat tgacaaggag gagggcacca
     8941 cacaaaaagt taggtgtaac agaaaatcat gaaact
//
