LOCUS       sequence-319721-        7765 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses OLE1 under the control of the yeast Sup35 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7765)
  AUTHORS   Sienski G, Narayan P, Bonner JM, Kory N, Boland S, Arczewska AA, 
            Ralvenius WT, Akay L, Lockshin E, He L, Milo B, Graziosi A, Baru V, 
            Lewis CA, Kellis M, Sabatini DM, Tsai LH, Lindquist S
  TITLE     APOE4 disrupts intracellular lipid homeostasis in human iPSC-derived
            glia.
  JOURNAL   Sci Transl Med. 2021 Mar 3;13(583). pii: 13/583/eaaz4564. doi: 
            10.1126/scitranslmed.aaz4564.
  PUBMED    33658354
REFERENCE   2  (bases 1 to 7765)
  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: "Sci Transl 
            Med."; date: "2021-03-3"; volume: "13(583). pii: 13/583/eaaz4564. 
            doi"; pages: " 10.1126/scitranslmed.aaz4564"
COMMENT     Sequence Label: pGS_591
FEATURES             Location/Qualifiers
     source          1..7765
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(458..673)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(698..716)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        784..888
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(889..957,958..1749)
                     /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(1107..1126)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1920..2508
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2409..2428
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2662..2679
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2796..2817
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2832..2837,2838..2855,2856..2862)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2870..2886
                     /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     2875..2897
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2894..2910
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2894..2910
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5149..5166)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5149..5165)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5158..5180)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    5922..7084
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN4 centromere fused to the 
                     autonomously replicating sequence ARS1/ARS416"
     rep_origin      6247..7084
                     /label=ARS1
                     /note="S. cerevisiae autonomously replicating sequence 
                     ARS1/ARS416"
     CDS             complement(join(7419..7765,1..457))
                     /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"
ORIGIN
        1 attctgctaa catcaaaagg cctctaggtt cctttgttac ttcttctgcc gcctgcttca
       61 aaccgctaac aatacctggg cccaccacac cgtgtgcatt cgtaatgtct gcccattctg
      121 ctattctgta tacacccgca gagtactgca atttgactgt attaccaatg tcagcaaatt
      181 ttctgtcttc gaagagtaaa aaattgtact tggcggataa tgcctttagc ggcttaactg
      241 tgccctccat ggaaaaatca gtcaagatat ccacatgtgt ttttagtaaa caaattttgg
      301 gacctaatgc ttcaactaac tccagtaatt ccttggtggt acgaacatcc aatgaagcac
      361 acaagtttgt ttgcttttcg tgcatgatat taaatagctt ggcagcaaca ggactaggat
      421 gagtagcagc acgttcctta tatgtagctt tcgacatgat ttatcttcgt ttcctgcatg
      481 tttttgttct gtgcagttgg gttaagaata ctgggcaatt tcatgtttct tcaacactac
      541 atatgcgtat atataccaat ctaagtctgt gctccttcct tcgttcttcc ttctgttcgg
      601 agattaccga atcaaaaaaa tttcaaggaa accgaaatca aaaaaaagaa taaaaaaaaa
      661 atgatgaatt gaaaagctaa ttcttgaaga cgaaagggcc tcgtgatacg cctattttta
      721 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
      781 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
      841 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
      901 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
      961 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     1021 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     1081 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     1141 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     1201 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     1261 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     1321 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     1381 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     1441 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     1501 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     1561 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     1621 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     1681 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     1741 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     1801 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     1861 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     1921 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     1981 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     2041 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     2101 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     2161 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     2221 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     2281 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     2341 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     2401 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     2461 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     2521 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     2581 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     2641 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     2701 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     2761 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     2821 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     2881 taacaatttc acacaggaaa cagctatgac catgattacg ccacaaccac acaaaaatca
     2941 tacaacgaat ggtatatgct tcatttcttt gtttcgcatt agctgcgcta tttgactcaa
     3001 attattattt tttactaaga cgacgcgtca cagtgttcga gtctgtgtca tttcttttgt
     3061 aattctctta aaccacttca taaagttgtg aagttcatag caaaattctt ccgcaaaaag
     3121 atgaatctta gttctcagcc caccaaaaga ggtacatgct aagatcatac agaagttatt
     3181 gtcacttctt accttgctct taaatgtaca ttacaaccgg gtattatatc ttacatcatc
     3241 gtataatatg atctttcttt atggagaaaa tttttttttt cactcgacca aagctcccat
     3301 tgcttctgaa gagtgtagtg tatattggta catcttctct tgaaagactc cattgtactg
     3361 taacaaaaag cggtttcttc atcgacttgc tcggaataac atctatatct gcccactagc
     3421 aacaaaaaca actaattcga aatgccaact tctggaacta ctattgaatt gattgacgac
     3481 caatttccaa aggatgactc tgccagcagt ggcattgtcg acgaagtcga cttaacggaa
     3541 gctaatattt tggctactgg tttgaataag aaagcaccaa gaattgtcaa cggttttggt
     3601 tctttaatgg gctccaagga aatggtttcc gtggaattcg acaagaaggg aaacgaaaag
     3661 aagtccaatt tggatcgtct gctagaaaag gacaaccaag aaaaagaaga agctaaaact
     3721 aaaattcaca tctccgaaca accatggact ttgaataact ggcaccaaca tttgaactgg
     3781 ttgaacatgg ttcttgtttg tggtatgcca atgattggtt ggtactttgc tctctctggt
     3841 aaagtgcctt tgcatttaaa cgttttcctt ttctccgttt tctactacgc tgtcggtggt
     3901 gtttctatta ctgccggtta ccatagatta tggtctcaca gatcttactc cgctcactgg
     3961 ccattgagat tattctacgc tatcttcggt tgtgcttccg ttgaagggtc cgctaaatgg
     4021 tggggccact ctcacagaat tcaccatcgt tacactgata ccttgagaga tccttatgac
     4081 gctcgtagag gtctatggta ctcccacatg ggatggatgc ttttgaagcc aaatccaaaa
     4141 tacaaggcta gagctgatat taccgatatg actgatgatt ggaccattag attccaacac
     4201 agacactaca tcttgttgat gttgttaacc gctttcgtca ttccaactct tatctgtggt
     4261 tactttttca acgactatat gggtggtttg atctatgccg gttttattcg tgtctttgtc
     4321 attcaacaag ctaccttttg cattaactcc ttggctcatt acatcggtac ccaaccattc
     4381 gatgacagaa gaacccctcg tgacaactgg attactgcca ttgttacttt cggtgaaggt
     4441 taccataact tccaccacga attcccaact gattacagaa acgctattaa gtggtaccaa
     4501 tacgacccaa ctaaggttat catctatttg acttctttag ttggtctagc atacgacttg
     4561 aagaaattct ctcaaaatgc tattgaagaa gccttgattc aacaagaaca aaagaagatc
     4621 aataaaaaga aggctaagat taactggggt ccagttttga ctgatttgcc aatgtgggac
     4681 aaacaaacct tcttggctaa gtctaaggaa aacaagggtt tggttatcat ttctggtatt
     4741 gttcacgacg tatctggtta tatctctgaa catccaggtg gtgaaacttt aattaaaact
     4801 gcattaggta aggacgctac caaggctttc agtggtggtg tctaccgtca ctcaaatgcc
     4861 gctcaaaatg tcttggctga tatgagagtg gctgttatca aggaaagtaa gaactctgct
     4921 attagaatgg ctagtaagag aggtgaaatc tacgaaactg gtaagttctt ttaaggtgga
     4981 gctctaagca aatagctaaa ttatatacga attaatatta tgattaagtg tttacgtgag
     5041 tgcgatattt ttattactat cttatacagt tgtatatact ctataaaatg agttgtctat
     5101 taattaacgc gatgaatgct ttctggcccg ggtaccgagc tcgaattcac tggccgtcgt
     5161 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     5221 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     5281 gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat tttctcctta cgcatctgtg
     5341 cggtatttca caccgcataa tcgctgggcc attctcatga agaatatctt gaatttattg
     5401 tcatattact agttggtgtg gaagtcctaa tatcggtgat caatatagtg gttgacatgc
     5461 tggctagtca acattgagcc ttttgatcat gcaaatatat tacggtattt tacaatcaaa
     5521 tatcaaactt aactattgac tttataactt atttaggtgg taacattctt ataaaaaaga
     5581 aaaaaattac tgcaaaacag tactagcttt taacttgtat cctaggttat ctatgctgtc
     5641 tcaccataga gaatattacc tatttcagaa tgtatgtcca tgattcgccg ggtaaataca
     5701 tataatacac aaatctggct taataaagtc tataatatat ctcataaaga agtgctaaat
     5761 tggctagtgc tatatatttt taagaaaatt tcttttgact aagtccatat cgactttgta
     5821 aaagttcaca ttagcataca tatattacac gagccagaaa tagtaacttt tgcctaaatc
     5881 acaaattgca aaatttaatt gcttgcaaaa ggtcacatgc ttataatcaa cttttttaaa
     5941 aatttaaaat acttttttat tttttatttt taaacataaa tgaaataatt tatttattgt
     6001 ttatgattac cgaaacataa aacctgctca agaaaaagaa actgttttgt ccttggaaaa
     6061 aaagcactac ctaggagcgg ccaaaatgcc gaggctttca tagcttaaac tctttacaga
     6121 aaataggcat tatagatcag ttcgagtttt cttattcttc cttccggttt tatcgtcaca
     6181 gttttacagt aaataagtat cacctcttag agttcgatga taagctgtca aacatgagaa
     6241 ttaattccac atgttaaaat agtgaaggag catgttcggc acacagtgga ccgaacgtgg
     6301 ggtaagtgca ctagggtccg gttaaacgga tctcgcattg atgaggcaac gctaattatc
     6361 aacatataga ttgttatcta tctgcatgaa cacgaaatct ttacttgacg acttgaggct
     6421 gatggtgttt atgcaaagaa accactgtgt ttaatatgtg tcactgtttg atattactgt
     6481 cagcgtagaa gataatagta aaagcggtta ataagtgtat ttgagataag tgtgataaag
     6541 tttttacagc gaaaagacga taaatacaag aaaatgatta cgaggatacg gagagaggta
     6601 tgtacatgtg tatttatata ctaagctgcc ggcggttgtt tgcaagaccg agaaaaggct
     6661 agcaagaatc gggtcattgt agcgtatgcg cctgtgaaca ttctcttcaa caagtttgat
     6721 tccattgcgg tgaaatggta aaagtcaacc ccctgcgatg tatattttcc tgtacaatca
     6781 atcaaaaagc caaatgattt agcattatct ttacatcttg ttattttaca gattttatgt
     6841 ttagatcttt tatgcttgct tttcaaaagg cttgcaggca agtgcacaaa caatacttaa
     6901 ataaatacta ctcagtaata acctatttct tagcattttt gacgaaattt gctattttgt
     6961 tagagtcttt tacaccattt gtctccacac ctccgcttac atcaacacca ataacgccat
     7021 ttaatctaag cgcatcacca acattttctg gcgtcagtcc accagctaac ataaaatgta
     7081 agctctcggg gctctcttgc cttccaaccc agtcagaaat cgagttccaa tccaaaagtt
     7141 cacctgtccc acctgcttct gaatcaaaca agggaataaa cgaatgaggt ttctgtgaag
     7201 ctgcactgag tagtatgttg cagtcttttg gaaatacgag tcttttaata actggcaaac
     7261 cgaggaactc ttggtattct tgccacgact catctccatg cagttggacg atcgatgata
     7321 agctgtcaaa catgagaatt gggtaataac tgatataatt aaattgaagc tctaatttgt
     7381 gagtttagta tacatgcatt tacttataat acagtttttt agttttgctg gccgcatctt
     7441 ctcaaatatg cttcccagcc tgcttttctg taacgttcac cctctacctt agcatccctt
     7501 ccctttgcaa atagtcctct tccaacaata ataatgtcag atcctgtaga gaccacatca
     7561 tccacggttc tatactgttg acccaatgcg tctcccttgt catctaaacc cacaccgggt
     7621 gtcataatca accaatcgta accttcatct cttccaccca tgtctctttg agcaataaag
     7681 ccgataacaa aatctttgtc gctcttcgca atgtcaacag tacccttagt atattctcca
     7741 gtagataggg agcccttgca tgaca
//
