LOCUS       sequence-319716-        8763 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Human APOE4 with a yeast secretory signal peptide under 
            the control of the yeast Gal promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8763)
  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 8763)
  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_101
FEATURES             Location/Qualifiers
     source          1..8763
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1855..2296
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    1855..1972
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     2267..2290
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     complement(4627..4646)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4628..4646)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4653..4670)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4653..4669)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4662..4684)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4810..5265
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4897..4916)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5107..5128
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5621..5640)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5740..5762
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5800..5818)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5903..6007
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6008..6076,6077..6868)
                     /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(6226..6245)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7039..7627
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7528..7547
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7781..7798
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7915..7936
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7951..7956,7957..7974,7975..7981)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7989..8005
                     /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     7994..8016
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8013..8029
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8013..8029
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        8050..8068
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        8356..8763
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
ORIGIN
        1 atgtctgccc ctaagttacc aatagatagc ggtttaatat aaggtttcaa caagaaatat
       61 tttgtgaaaa tcaaaagtta atagtttcac tgtcgaaaat atgaaagttt aagttttcag
      121 ttcatcccat tgtagttatc aaatttctgt gtttcccgta tatgtgtgct ctgtattcat
      181 cgctatcttg aaactggtta tgatcgttgc gcatgtgacc tgtatcgctc aaaagagccc
      241 ggcaccggta ccgggcaaaa gcccctagca agctaaaatg attcaacttt cgtagtaatt
      301 tgttcaaccg gtcacagtaa agtcaatgac taacttttga atcgaggatg tatttccaat
      361 tgaaagcaat gtatatataa accattgtgt tgaagcttgt cttacagttt tcttgtatcc
      421 tcccattgtt catatagtac ttaaagtacc ataatcctta aagaacaaac ttgaaagaga
      481 atacactata cacaactaat aaataaagaa taatcatggc tgtgaccaag acaattgtag
      541 ttctaccagg tgaccatgtt ggtcaagaaa tcactgaaga ggccattaaa gttttaaatg
      601 ctattcagga atgtcgtcca gacaaggtca atttcaagtt tgatcatcat ttgatcggtg
      661 gtgctgcaat tgatgccact ggtgttccat taccagacga agctttggag gcctctaaga
      721 aagctgatgc cgtgcttctg ggtgctgttg gtggtccaaa atggggtact ggcgctgtca
      781 gaccagaaca aggtcttttg aaaatccgta aggagttgca attgtatgct aatctaagac
      841 catgtaattt tgcatctgat tccttactag atctatcgcc attgaagcct gaaattgcaa
      901 gaggtacaga tttcgtcgtt gttagagaac tagtgggtgg tatttatttc ggggagagaa
      961 aagaagatga aggtgatggt gtcgcctggg atagcgaaaa gtattctgtg cccgaagttc
     1021 aaagaatcac tagaatggct gctttcatgg ctctgcaaca caacccacct ttacctattt
     1081 ggtctctaga taaagccaac gttctagcat catcaagatt atggagaaaa acagtggaag
     1141 agacgattaa gaatgaattc ccacaattga ctgttcaaca tcaattgatt gattccgctg
     1201 ctatgatctt ggttaagaac ccaactcatc taaatggtat tattatcaca aacaatatgt
     1261 ttggtgatat tatctccgat gaagcctccg tgatcccagg ttcgttagga ttacttccat
     1321 ctgcctcctt ggcatctctt ccagataaga atacggcatt tggtttatat gagccatgtc
     1381 atggctctgc tccagactta ccaaagggaa aggttaaccc cgtagcaaca atcctatctg
     1441 ctgctatgat gttgaagttg tctctggacc tgttcgaaga aggtgaaatt atagaacaag
     1501 cagtcaagaa agtgttggat tctggtatca gaacagcgga tctaaagggt accaactcta
     1561 ctactgaggt cggtgatgca gtagcgaaag ctgtcaggga actattagct tagacttagt
     1621 agatgtacag ttgcatacat aagtctgaac agaaagaagt aaatatcata tctagttcgg
     1681 tttagaaatc agtgactata ttggttttga ccagatcgca atccttgcaa gcatttagta
     1741 atgactatca aaccggtgaa tcatctgata aggctatatt agcttgtgca ttcgcatgta
     1801 tcgggaaacg aacggtaccg ggcccaaata ccaacaaaag ctggagctct agtacggatt
     1861 agaagccgcc gagcgggtga cagccctccg aaggaagact ctcctccgtg cgtcctcgtc
     1921 ttcaccggtc gcgttcctga aacgcagatg tgcctcgcgc cgcactgctc cgaacaataa
     1981 agattctaca atactagctt ttatggttat gaagaggaaa aattggcagt aacctggccc
     2041 cacaaacctt caaatgaacg aatcaaatta acaaccatag gatgataatg cgattagttt
     2101 tttagcctta tttctggggt aattaatcag cgaagcgatg atttttgatc tattaacaga
     2161 tatataaatg caaaaactgc ataaccactt taactaatac tttcaacatt ttcggtttgt
     2221 attacttctt attcaaatgt aataaaagta tcaacaaaaa attgttaata tacctctata
     2281 ctttaacgtc aaggagaaaa aaccaaaaca actaattcga aatgtttttc aacagactaa
     2341 gcgctggcaa gctgctggta ccactctccg tggtcctgta cgcccttttc gtggtaatat
     2401 tacctttaca gaattctttc cactcctcca atgttttagt tagaggtaag gtggagcaag
     2461 cggtggagac agagccggag cccgagctgc gccagcagac cgagtggcag agcggccagc
     2521 gctgggaact ggcactgggt cgcttttggg attacctgcg ctgggtgcag acactgtctg
     2581 agcaggtgca ggaggagctg ctcagctccc aggtcaccca ggaactgagg gcgctgatgg
     2641 acgagaccat gaaggagttg aaggcctaca aatcggaact ggaggaacaa ctgaccccgg
     2701 tggcggagga gacgcgggca cggctgtcca aggagctgca ggcggcgcag gcccggctgg
     2761 gcgcggacat ggaggacgtg cgcggccgcc tggtgcagta ccgcggcgag gtgcaggcca
     2821 tgctcggcca gagcaccgag gagctgcggg tgcgcctcgc ctcccacctg cgcaagctgc
     2881 gtaagcggct cctccgcgat gccgatgacc tgcagaagcg cctggcagtg taccaggccg
     2941 gggcccgcga gggcgccgag cgcggcctca gcgccatccg cgagcgcctg gggcccctgg
     3001 tggaacaggg ccgcgtgcgg gccgccactg tgggctccct ggccggccag ccgctacagg
     3061 agcgggccca ggcctggggc gagcggctgc gcgcgcggat ggaggagatg ggcagccgga
     3121 cccgcgaccg cctggacgag gtgaaggagc aggtggcgga ggtgcgcgcc aagctggagg
     3181 agcaggccca gcagatacgc ctgcaggccg aggccttcca ggcccgcctc aagagctggt
     3241 tcgagcccct ggtggaagac atgcagcgcc agtgggccgg gctggtggag aaggtgcagg
     3301 ctgccgtggg caccagcgcc gcccctgtgc ccagcgacaa tcactgaggc cgccaccgcg
     3361 gtggagctct aagcaaatag ctaaattata tacgaattaa tattatgatt aagtgtttac
     3421 gtgagtgcga tatttttatt actatcttat acagttgtat atactctata aaatgagttg
     3481 tctattaatt aacgcgatga atgctttctg ggtttacctc tccaacaact ctagtttact
     3541 tctcaataca ttcaattgta tttgatttgt caatacttca tcattaatca attctatagt
     3601 tttgtttttc tcgtttattt ccaaatttaa tgcatcaatt ttattattca atttgtcgtt
     3661 gattttggtt aatgatttta tggtttgatc tctggcattg attgtttgtg ttagtttttc
     3721 attattgata attaaattat ttaagttagt tatcaactcg gtgttttcaa gtttcaagtt
     3781 ttcaatttct ttagagttta ttagatttgt caaagtttct gaattgcttg attggtcctg
     3841 tagaagagta tttgttgttg tggataattg attcaatttt tgagacaatt gctggaaggc
     3901 gttgaaatat ctagcatcaa tctcatggtt tttttcccga gagtctcgta gattcaattg
     3961 ttttaatata tcttgggacc actcttgatt tgaactcatg gaaattaaac tgggtgttgt
     4021 gttgtggtgt aatgattgta ccccctttgc ttataattgt gtggccaccg aagtcggtga
     4081 tgctgtcgcc gaagaagtta agaaaatcct tgcttaaaaa gattctcttt ttttatgata
     4141 tttgtacata aactttataa atgaaattca taatagaaac gacacgaaat tacaaaatgg
     4201 aatatgttca tagggtagac gaaactatat acgcaatcta catacattta tcaagaagga
     4261 gaaaaaggag gatgtaaagg aatacaggta agcaaattga tactaatggc tcaacgtgat
     4321 aaggaaaaag aattgcactt taacattaat attgacaagg aggagggcac cacacaaaaa
     4381 gttaggtgta acagaaaatc atgaaactat gattcctaat ttatatattg gaggattttc
     4441 tctaaaaaaa aaaaaataca acaaataaaa aacactcaat gacctgacca tttgatggag
     4501 tttaagtcaa tacctccttg aaccatttcc cataatggtg aaagttccct caagaatttt
     4561 actctgtcag aaacggcctt aacgacgtag gccggccatt taaatgttta aacggccacg
     4621 ctggccccct atagtgagtc gtattacaat tcactggccg tcgttttaca acgtcgtgac
     4681 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     4741 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     4801 ggcgaatgga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     4861 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     4921 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     4981 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     5041 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     5101 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     5161 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     5221 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttcctga tgcggtattt
     5281 tctccttacg catctgtgcg gtatttcaca ccgcataggg taataactga tataattaaa
     5341 ttgaagctct aatttgtgag tttagtatta ggatctctac ccttggcgaa aagtcctctg
     5401 ccaacaatga tgatatctga tccaccactt acaacttcgt cgacggttct gtactgctga
     5461 cccaatatgc gtatatatac caatctaagt ctgtgctcct tccttcgttc ttccttctgt
     5521 tcggagatta ccgaatcaaa aaaatttcaa ggaaaccgaa atcaaaaaaa agaataaaaa
     5581 aaaaatgatg aattgaaaag gtggtatggt gcactctcag tacaatctgc tctgatgccg
     5641 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     5701 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     5761 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     5821 tataggttaa tgtcatgata ataatggttt cttagacgtc ggcgcgccga ggacgtcagg
     5881 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     5941 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     6001 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     6061 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     6121 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     6181 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     6241 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     6301 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     6361 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     6421 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     6481 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     6541 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     6601 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     6661 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     6721 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     6781 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     6841 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     6901 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     6961 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     7021 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     7081 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     7141 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     7201 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     7261 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     7321 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     7381 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     7441 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     7501 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     7561 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     7621 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     7681 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     7741 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     7801 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     7861 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     7921 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     7981 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
     8041 caagctcgga attaaccctc actaaaggga acaaaagctg ggccggccat ttaaatgttt
     8101 aaacggccac gctggccgag gagaacttct agtatatcta catacctaat attattgcct
     8161 tattaaaaat ggaatcccaa cgattacatc aaaatccaca ttctcttcaa aatcaattgt
     8221 cctgtacttc cttgttcatg tgtgttcaaa aacgttatat ttataggata attatactct
     8281 atttctcaac aagtaattgg ttgtttggcc gagcggtcta aggcgcctga ttcaagaaat
     8341 atcttgaccg cagttaactg tgggaatact caggtatcgt aagatgcaag agttcgaatc
     8401 tcttagcaac cattattttt ttcctcaaca taacgagaac acacaggggc gctatcgcac
     8461 agaatcaaat tcgatgactg gaaatttttt gttaatttca gaggtcgcct gacgcatata
     8521 cctttttcaa ctgaaaaatt gggagaaaaa ggaaaggtga gagcgccgga accggctttt
     8581 catatagaat agagaagcgt tcatgactaa atgcttgcat cacaatactt gaagttgaca
     8641 atattattta aggacctatt gttttttcca ataggtggtt agcaatcgtc ttactttcta
     8701 acttttctta ccttttacat ttcagcaata tatatatata tatttcaagg atataccatt
     8761 cta
//
