LOCUS       sequence-319714-        8973 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Human APOE3 with a yeast secretory signal peptide under 
            the control of a beta-estradiol inducible promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8973)
  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 8973)
  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_128
FEATURES             Location/Qualifiers
     source          1..8973
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     2483..2506
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     complement(4837..4856)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4838..4856)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4863..4880)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4863..4879)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4872..4894)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5020..5475
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5107..5126)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5317..5338
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5831..5850)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5950..5972
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6010..6028)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6113..6217
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6218..6286,6287..7078)
                     /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(6436..6455)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7249..7837
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7738..7757
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7991..8008
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8125..8146
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8161..8166,8167..8184,8185..8191)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8199..8215
                     /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     8204..8226
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8223..8239
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8223..8239
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        8260..8278
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        8566..8973
                     /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 gcccttatat tgaattttca aaaattctta cttttttttt
     1861 ggatggacgc aaagaagttt aataatcata ttacatggca ttaccaccat atacatatcc
     1921 atatacatat ccatatctaa tcttacttat atgttgtgga aatgtaaaga gccccattat
     1981 cttagcctaa aaaaaccttc tctttggaac tttcagtaat acgcttaact gctcattgct
     2041 atattgaagt gcggccgcgt gggcgtgcgt gggcgggcgt gggcgtgcgt gggcgggcgt
     2101 gggcgtgcgt gggcgtctag accgtgcgtc ctcgtcttca ccggtcgcgt tcctgaaacg
     2161 cagatgtgcc taatgccgca ctgctccgaa caataaagat tctacaatac tagcttttat
     2221 ggttatgaag aggaaaaatt ggcagtaacc tggccccaca aaccttcaaa ttaacgaatc
     2281 aaattaacaa ccataggatg ataatgcgat tagtttttta gccttatttc tggggtaatt
     2341 aatcagcgaa gcgatgattt ttgatctatt aacagatata taaatggaaa agctgcataa
     2401 ccactttaac taatactttc aacattttca gtttgtatta cttcttattc aaatgtcata
     2461 aaagtatcaa caaaaaattg ttaatatacc tctatacttt aacgtcaagg agaaaaaact
     2521 atactcgagt aacaaaatgt ttttcaacag actaagcgct ggcaagctgc tggtaccact
     2581 ctccgtggtc ctgtacgccc ttttcgtggt aatattacct ttacagaatt ctttccactc
     2641 ctccaatgtt ttagttagag gtaaggtgga gcaagcggtg gagacagagc cggagcccga
     2701 gctgcgccag cagaccgagt ggcagagcgg ccagcgctgg gaactggcac tgggtcgctt
     2761 ttgggattac ctgcgctggg tgcagacact gtctgagcag gtgcaggagg agctgctcag
     2821 ctcccaggtc acccaggaac tgagggcgct gatggacgag accatgaagg agttgaaggc
     2881 ctacaaatcg gaactggagg aacaactgac cccggtggcg gaggagacgc gggcacggct
     2941 gtccaaggag ctgcaggcgg cgcaggcccg gctgggcgcg gacatggagg acgtgtgcgg
     3001 ccgcctggtg cagtaccgcg gcgaggtgca ggccatgctc ggccagagca ccgaggagct
     3061 gcgggtgcgc ctcgcctccc acctgcgcaa gctgcgtaag cggctcctcc gcgatgccga
     3121 tgacctgcag aagcgcctgg cagtgtacca ggccggggcc cgcgagggcg ccgagcgcgg
     3181 cctcagcgcc atccgcgagc gcctggggcc cctggtggaa cagggccgcg tgcgggccgc
     3241 cactgtgggc tccctggccg gccagccgct acaggagcgg gcccaggcct ggggcgagcg
     3301 gctgcgcgcg cggatggagg agatgggcag ccggacccgc gaccgcctgg acgaggtgaa
     3361 ggagcaggtg gcggaggtgc gcgccaagct ggaggagcag gcccagcaga tacgcctgca
     3421 ggccgaggcc ttccaggccc gcctcaagag ctggttcgag cccctggtgg aagacatgca
     3481 gcgccagtgg gccgggctgg tggagaaggt gcaggctgcc gtgggcacca gcgccgcccc
     3541 tgtgcccagc gacaatcact gataaccgcg gtggagctct aagcaaatag ctaaattata
     3601 tacgaattaa tattatgatt aagtgtttac gtgagtgcga tatttttatt actatcttat
     3661 acagttgtat atactctata aaatgagttg tctattaatt aacgcgatga atgctttctg
     3721 ggtttacctc tccaacaact ctagtttact tctcaataca ttcaattgta tttgatttgt
     3781 caatacttca tcattaatca attctatagt tttgtttttc tcgtttattt ccaaatttaa
     3841 tgcatcaatt ttattattca atttgtcgtt gattttggtt aatgatttta tggtttgatc
     3901 tctggcattg attgtttgtg ttagtttttc attattgata attaaattat ttaagttagt
     3961 tatcaactcg gtgttttcaa gtttcaagtt ttcaatttct ttagagttta ttagatttgt
     4021 caaagtttct gaattgcttg attggtcctg tagaagagta tttgttgttg tggataattg
     4081 attcaatttt tgagacaatt gctggaaggc gttgaaatat ctagcatcaa tctcatggtt
     4141 tttttcccga gagtctcgta gattcaattg ttttaatata tcttgggacc actcttgatt
     4201 tgaactcatg gaaattaaac tgggtgttgt gttgtggtgt aatgattgta ccccctttgc
     4261 ttataattgt gtggccaccg aagtcggtga tgctgtcgcc gaagaagtta agaaaatcct
     4321 tgcttaaaaa gattctcttt ttttatgata tttgtacata aactttataa atgaaattca
     4381 taatagaaac gacacgaaat tacaaaatgg aatatgttca tagggtagac gaaactatat
     4441 acgcaatcta catacattta tcaagaagga gaaaaaggag gatgtaaagg aatacaggta
     4501 agcaaattga tactaatggc tcaacgtgat aaggaaaaag aattgcactt taacattaat
     4561 attgacaagg aggagggcac cacacaaaaa gttaggtgta acagaaaatc atgaaactat
     4621 gattcctaat ttatatattg gaggattttc tctaaaaaaa aaaaaataca acaaataaaa
     4681 aacactcaat gacctgacca tttgatggag tttaagtcaa tacctccttg aaccatttcc
     4741 cataatggtg aaagttccct caagaatttt actctgtcag aaacggcctt aacgacgtag
     4801 gccggccatt taaatgttta aacggccacg ctggccccct atagtgagtc gtattacaat
     4861 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     4921 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     4981 cgcccttccc aacagttgcg cagcctgaat ggcgaatgga cgcgccctgt agcggcgcat
     5041 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     5101 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     5161 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     5221 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     5281 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     5341 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     5401 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     5461 taacgcttac aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     5521 ccgcataggg taataactga tataattaaa ttgaagctct aatttgtgag tttagtatta
     5581 ggatctctac ccttggcgaa aagtcctctg ccaacaatga tgatatctga tccaccactt
     5641 acaacttcgt cgacggttct gtactgctga cccaatatgc gtatatatac caatctaagt
     5701 ctgtgctcct tccttcgttc ttccttctgt tcggagatta ccgaatcaaa aaaatttcaa
     5761 ggaaaccgaa atcaaaaaaa agaataaaaa aaaaatgatg aattgaaaag gtggtatggt
     5821 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     5881 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     5941 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     6001 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     6061 cttagacgtc ggcgcgccga ggacgtcagg tggcactttt cggggaaatg tgcgcggaac
     6121 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     6181 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     6241 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     6301 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     6361 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     6421 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     6481 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     6541 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     6601 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     6661 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     6721 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     6781 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     6841 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     6901 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     6961 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     7021 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     7081 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     7141 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     7201 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     7261 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     7321 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     7381 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     7441 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     7501 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     7561 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     7621 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     7681 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     7741 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     7801 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     7861 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     7921 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     7981 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     8041 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     8101 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
     8161 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     8221 cacaggaaac agctatgacc atgattacgc caagctcgga attaaccctc actaaaggga
     8281 acaaaagctg ggccggccat ttaaatgttt aaacggccac gctggccgag gagaacttct
     8341 agtatatcta catacctaat attattgcct tattaaaaat ggaatcccaa cgattacatc
     8401 aaaatccaca ttctcttcaa aatcaattgt cctgtacttc cttgttcatg tgtgttcaaa
     8461 aacgttatat ttataggata attatactct atttctcaac aagtaattgg ttgtttggcc
     8521 gagcggtcta aggcgcctga ttcaagaaat atcttgaccg cagttaactg tgggaatact
     8581 caggtatcgt aagatgcaag agttcgaatc tcttagcaac cattattttt ttcctcaaca
     8641 taacgagaac acacaggggc gctatcgcac agaatcaaat tcgatgactg gaaatttttt
     8701 gttaatttca gaggtcgcct gacgcatata cctttttcaa ctgaaaaatt gggagaaaaa
     8761 ggaaaggtga gagcgccgga accggctttt catatagaat agagaagcgt tcatgactaa
     8821 atgcttgcat cacaatactt gaagttgaca atattattta aggacctatt gttttttcca
     8881 ataggtggtt agcaatcgtc ttactttcta acttttctta ccttttacat ttcagcaata
     8941 tatatatata tatttcaagg atataccatt cta
//
