LOCUS       sequence-417212-        8976 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the misfolded CPY* mutant 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(172..191)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     291..313
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(351..369)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    411..914
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        951..1055
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1056..1124,1125..1916)
                     /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(1274..1293)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2087..2675
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2576..2595
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2829..2846
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2963..2984
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2999..3004,3005..3022,3023..3029)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3037..3053
                     /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     3042..3064
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3061..3077
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3061..3077
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3096..3116
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3098..3116
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3140..3581
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    3140..3257
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     3552..3575
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     terminator      5945..6192
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5961..5979)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(6210..6229)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6211..6229)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6239..6256)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6239..6255)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6248..6270)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6396..6851
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6483..6502)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6693..6714
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7152..7559
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             7560..8654
                     /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 ctctaaaaaa aaaaaaatac aacaaataaa aaacactcaa tgacctgacc atttgatgga
       61 gtttaagtca ataccttctt gaaccatttc ccataatggt gaaagttccc tcaagaattt
      121 tactctgtca gaaacggcct taacgacgta gtcgatatgg tgcactctca gtacaatctg
      181 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
      241 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
      301 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
      361 acgcctattt ttataggtta atgtcatgat aataatggtt tcttaggacg gatcgcttgc
      421 ctgtaactta cacgcgcctc gtatctttta atgatggaat aatttgggaa tttactctgt
      481 gtttatttat ttttatgttt tgtatttgga ttttagaaag taaataaaga aggtagaaga
      541 gttacggaat gaagaaaaaa aaataaacaa aggtttaaaa aatttcaaca aaaagcgtac
      601 tttacatata tatttattag acaagaaaag cagattaaat agatatacat tcgattaacg
      661 ataagtaaaa tgtaaaatca caggattttc gtgtgtggtc ttctacacag acaagatgaa
      721 acaattcggc attaatacct gagagcagga agagcaagat aaaaggtagt atttgttggc
      781 gatcccccta gagtctttta catcttcgga aaacaaaaac tattttttct ttaatttctt
      841 tttttacttt ctatttttaa tttatatatt tatattaaaa aatttaaatt ataattattt
      901 ttatagcacg tgatgaaaag gacccaggtg gcacttttcg gggaaatgtg cgcggaaccc
      961 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     1021 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     1081 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     1141 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     1201 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     1261 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     1321 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     1381 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     1441 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     1501 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     1561 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     1621 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     1681 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     1741 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     1801 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     1861 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     1921 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     1981 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     2041 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     2101 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     2161 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     2221 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     2281 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     2341 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     2401 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     2461 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     2521 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     2581 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     2641 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     2701 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     2761 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     2821 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     2881 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     2941 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
     3001 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     3061 caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac
     3121 aaaagctgga gctctagtac ggattagaag ccgccgagcg ggtgacagcc ctccgaagga
     3181 agactctcct ccgtgcgtcc tcgtcttcac cggtcgcgtt cctgaaacgc agatgtgcct
     3241 cgcgccgcac tgctccgaac aataaagatt ctacaatact agcttttatg gttatgaaga
     3301 ggaaaaattg gcagtaacct ggccccacaa accttcaaat gaacgaatca aattaacaac
     3361 cataggatga taatgcgatt agttttttag ccttatttct ggggtaatta atcagcgaag
     3421 cgatgatttt tgatctatta acagatatat aaatgcaaaa actgcataac cactttaact
     3481 aatactttca acattttcgg tttgtattac ttcttattca aatgtaataa aagtatcaac
     3541 aaaaaattgt taatatacct ctatacttta acgtcaagga gaaaaaaccc cggattctag
     3601 aactagtcta aaaatgaaag cattcaccag tttactatgt ggactaggcc tgtccactac
     3661 actcgctaag gccatctcat tgcaaagacc gttgggtcta gataaggacg ttttgctgca
     3721 agctgcggaa aaatttggtt tggacctcga cctggatcat ctcttgaagg agttggactc
     3781 caatgtattg gacgcttggg cccaaataga gcatttgtac ccaaaccagg ttatgagcct
     3841 tgaaacttcc actaagccaa aattccctga agcaatcaaa acgaagaaag actgggactt
     3901 tgtggtcaag aatgacgcaa ttgaaaacta tcagcttcgt gtcaacaaga ttaaggaccc
     3961 taaaatcctg ggcattgacc caaatgtcac acagtacacg ggttacttgg atgtggaaga
     4021 cgaggacaag catttcttct tttggacttt tgaaagtaga aacgatcctg caaaggatcc
     4081 ggtcatcctt tggttgaacg ggggtccagg ttgttcttca ctaaccgggc tgttctttga
     4141 attaggaccc tcatccattg gacctgattt gaaacccatc gggaaccctt actcttggaa
     4201 cagcaatgcc accgtgatct tccttgacca gcctgtcaac gttgggttct cgtattccgg
     4261 gtcctcaggt gtttccaaca ctgtcgccgc tggtaaggat gtctataact tcttggagtt
     4321 gttcttcgat cagttccctg aatacgtcaa caagggccaa gatttccaca tcgctaggga
     4381 atcctacgcc ggccattaca tccctgtttt tgcctctgaa attttgtctc acaaggacag
     4441 aaacttcaac ttaacctccg tcttgatcgg aaatggcctc actgacccat tgactcagta
     4501 taactattac gaaccaatgg cctgtggtga aggtggcgaa ccatctgttt tgccctcgga
     4561 ggaatgctct gctatggaag actctttgga acgttgtttg ggcttgatcg agtcgtgcta
     4621 tgactcgcaa tccgtctggt cctgtgttcc agctaccatt tattgtaata acgcccaatt
     4681 ggctccttac caacgtaccg gcagaaacgt ttacgatatc aggaaggatt gtgaaggtgg
     4741 caatttgtgc tacccaacgt tacaagatat cgacgactac ttaaaccagg actacgtcaa
     4801 agaagctgtc ggtgcggagg ttgaccacta cgaatcctgt aacttcgata tcaacagaaa
     4861 tttcctgttt gcgggtgatt ggatgaagcc ttaccacacc gccgtaacag atcttttgaa
     4921 tcaagaccta cccattctgg tatatgcagg cgataaagat ttcatctgta actggttggg
     4981 taataaggcg tggacggatg tcttgccatg gaagtacgac gaagaatttg caagccaaaa
     5041 agtacgtaac tggactgctt ctatcaccga cgaggtcgct ggtgaagtca aatcctacaa
     5101 gcacttcacc tatttgagag tcttcaatgg tggccacatg gttccatttg acgtccctga
     5161 aaacgcctta agtatggtta acgaatggat ccacggtggt ttctccttaa agcttgtatc
     5221 caaaggtgaa gaattcttta ctggtgttgt tcctatttta gttgaattag acggtgacgt
     5281 taatggtcat aagtttagtg ttagaggtga aggtgaaggt gacgcaacta acggtaaatt
     5341 gacattgaag tttatttgta ctactggtaa attgccagtt ccatggccaa ccttggttac
     5401 cactttaact tatggtgttc aatgcttctc aagataccct gatcatatga aaagacacga
     5461 ctttttcaag agtgccatgc cagaaggtta cgtacaagaa agaactatat cttttaaaga
     5521 tgacggtaca tacaagacca gagctgaagt aaaattcgaa ggtgacacat tggtcaacag
     5581 aatcgaattg aagggtatcg atttcaagga agacggtaac atcttgggtc ataaattaga
     5641 atacaatttc aactcacaca atgtttacat cactgctgat aagcaaaaga atggtatcaa
     5701 agcaaacttc aagatcagac ataacgttga agatggttcc gtacaattag ctgaccacta
     5761 tcaacaaaat acacctattg gtgacggtcc tgttttgtta ccagacaacc attacttgtc
     5821 tacccaatca aaattgtcca aagatccaaa cgaaaagaga gaccatatgg tcttgttgga
     5881 atttgtcact gccgctggta tcactcacgg tatggatgaa ttgtataagt aggtcgacct
     5941 cgagtcatgt aattagttat gtcacgctta cattcacgcc ctccccccac atccgctcta
     6001 accgaaaagg aaggagttag acaacctgaa gtctaggtcc ctatttattt ttttatagtt
     6061 atgttagtat taagaacgtt atttatattt caaatttttc ttttttttct gtacagacgc
     6121 gtgtacgcat gtaacattat actgaaaacc ttgcttgaga aggttttggg acgctcgaag
     6181 gctttaattt gcggccggta cccaattcgc cctatagtga gtcgtattac gcgcgctcac
     6241 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     6301 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     6361 cttcccaaca gttgcgcagc ctgaatggcg aatggacgcg ccctgtagcg gcgcattaag
     6421 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     6481 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     6541 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     6601 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     6661 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     6721 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     6781 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     6841 gcttacaatt tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     6901 atatcgaccg gtcgaggaga acttctagta tatctacata cctaatatta ttgccttatt
     6961 aaaaatggaa tcccaacaat tacatcaaaa tccacattct cttcaaaatc aattgtcctg
     7021 tacttccttg ttcatgtgtg ttcaaaaacg ttatatttat aggataatta tactctattt
     7081 ctcaacaagt aattggttgt ttggccgagc ggtctaaggc gcctgattca agaaatatct
     7141 tgaccgcagt taactgtggg aatactcagg tatcgtaaga tgcaagagtt cgaatctctt
     7201 agcaaccatt atttttttcc tcaacataac gagaacacac aggggcgcta tcgcacagaa
     7261 tcaaattcga tgactggaaa ttttttgtta atttcagagg tcgcctgacg catatacctt
     7321 tttcaactga aaaattggga gaaaaaggaa aggtgagagc gccggaaccg gcttttcata
     7381 tagaatagag aagcgttcat gactaaatgc ttgcatcaca atacttgaag ttgacaatat
     7441 tatttaagga cctattgttt tttccaatag gtggttagca atcgtcttac tttctaactt
     7501 ttcttacctt ttacatttca gcaatatata tatatatatt tcaaggatat accattctaa
     7561 tgtctgcccc taagaagatc gtcgttttgc caggtgacca cgttggtcaa gaaatcacag
     7621 ccgaagccat taaggttctt aaagctattt ctgatgttcg ttccaatgtc aagttcgatt
     7681 tcgaaaatca tttaattggt ggtgctgcta tcgatgctac aggtgttcca cttccagatg
     7741 aggcgctgga agcctccaag aaggctgatg ccgttttgtt aggtgctgtg ggtggtccta
     7801 aatggggtac cggtagtgtt agacctgaac aaggtttact aaaaatccgt aaagaacttc
     7861 aattgtacgc caacttaaga ccatgtaact ttgcatccga ctctctttta gacttatctc
     7921 caatcaagcc acaatttgct aaaggtactg acttcgttgt tgtcagagaa ttagtgggag
     7981 gtatttactt tggtaagaga aaggaagacg atggtgatgg tgtcgcttgg gatagtgaac
     8041 aatacaccgt tccagaagtg caaagaatca caagaatggc cgctttcatg gccctacaac
     8101 atgagccacc attgcctatt tggtccttgg ataaagctaa tgttttggcc tcttcaagat
     8161 tatggagaaa aactgtggag gaaaccatca agaacgaatt ccctacattg aaggttcaac
     8221 atcaattgat tgattctgcc gccatgatcc tagttaagaa cccaacccac ctaaatggta
     8281 ttataatcac cagcaacatg tttggtgata tcatctccga tgaagcctcc gttatcccag
     8341 gttccttggg tttgttgcca tctgcgtcct tggcctcttt gccagacaag aacaccgcat
     8401 ttggtttgta cgaaccatgc cacggttctg ctccagattt gccaaagaat aaggtcaacc
     8461 ctatcgccac tatcttgtct gctgcaatga tgttgaaatt gtcattgaac ttgcctgaag
     8521 aaggtaaggc cattgaagat gcagttaaaa aggttttgga tgcaggtatc agaactggtg
     8581 atttaggtgg ttccaacagt accaccgaag tcggtgatgc tgtcgccgaa gaagttaaga
     8641 aaatccttgc ttaaaaagat tctctttttt tatgatattt gtacataaac tttataaatg
     8701 aaattcataa tagaaacgac acgaaattac aaaatggaat atgttcatag ggtagacgaa
     8761 actatatacg caatctacat acatttatca agaaggagaa aaaggaggat gtaaaggaat
     8821 acaggtaagc aaattgatac taatggctca acgtgataag gaaaaagaat tgcactttaa
     8881 cattaatatt gacaaggagg agggcaccac acaaaaagtt aggtgtaaca gaaaatcatg
     8941 aaactatgat tcctaattta tatattggag gatttt
//
