LOCUS       sequence-318526-        8973 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Human APOE4 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_129
FEATURES             Location/Qualifiers
     source          1..8973
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     4..26
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     18..35
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     19..35
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     42..61
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        42..60
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2392..2415)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     promoter        complement(4898..5305)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     promoter        complement(5593..5611)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5632..5648)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5632..5648)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5645..5667)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5656..5672
                     /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)."
     promoter        complement(join(5680..5686,5687..5704,5705..5710))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5725..5746
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5863..5880)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6034..6622)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6114..6133)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6793..7584,7585..7653))
                     /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     7416..7435
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7654..7758)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7843..7861
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7899..7921)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8021..8040
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(8396..8851)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8533..8554)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8745..8764
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt gtaatacgac tcactatagg
       61 gggccagcgt ggccgtttaa acatttaaat ggccggccta cgtcgttaag gccgtttctg
      121 acagagtaaa attcttgagg gaactttcac cattatggga aatggttcaa ggaggtattg
      181 acttaaactc catcaaatgg tcaggtcatt gagtgttttt tatttgttgt attttttttt
      241 ttttagagaa aatcctccaa tatataaatt aggaatcata gtttcatgat tttctgttac
      301 acctaacttt ttgtgtggtg ccctcctcct tgtcaatatt aatgttaaag tgcaattctt
      361 tttccttatc acgttgagcc attagtatca atttgcttac ctgtattcct ttacatcctc
      421 ctttttctcc ttcttgataa atgtatgtag attgcgtata tagtttcgtc taccctatga
      481 acatattcca ttttgtaatt tcgtgtcgtt tctattatga atttcattta taaagtttat
      541 gtacaaatat cataaaaaaa gagaatcttt ttaagcaagg attttcttaa cttcttcggc
      601 gacagcatca ccgacttcgg tggccacaca attataagca aagggggtac aatcattaca
      661 ccacaacaca acacccagtt taatttccat gagttcaaat caagagtggt cccaagatat
      721 attaaaacaa ttgaatctac gagactctcg ggaaaaaaac catgagattg atgctagata
      781 tttcaacgcc ttccagcaat tgtctcaaaa attgaatcaa ttatccacaa caacaaatac
      841 tcttctacag gaccaatcaa gcaattcaga aactttgaca aatctaataa actctaaaga
      901 aattgaaaac ttgaaacttg aaaacaccga gttgataact aacttaaata atttaattat
      961 caataatgaa aaactaacac aaacaatcaa tgccagagat caaaccataa aatcattaac
     1021 caaaatcaac gacaaattga ataataaaat tgatgcatta aatttggaaa taaacgagaa
     1081 aaacaaaact atagaattga ttaatgatga agtattgaca aatcaaatac aattgaatgt
     1141 attgagaagt aaactagagt tgttggagag gtaaacccag aaagcattca tcgcgttaat
     1201 taatagacaa ctcattttat agagtatata caactgtata agatagtaat aaaaatatcg
     1261 cactcacgta aacacttaat cataatatta attcgtatat aatttagcta tttgcttaga
     1321 gctccaccgc ggttatcagt gattgtcgct gggcacaggg gcggcgctgg tgcccacggc
     1381 agcctgcacc ttctccacca gcccggccca ctggcgctgc atgtcttcca ccaggggctc
     1441 gaaccagctc ttgaggcggg cctggaaggc ctcggcctgc aggcgtatct gctgggcctg
     1501 ctcctccagc ttggcgcgca cctccgccac ctgctccttc acctcgtcca ggcggtcgcg
     1561 ggtccggctg cccatctcct ccatccgcgc gcgcagccgc tcgccccagg cctgggcccg
     1621 ctcctgtagc ggctggccgg ccagggagcc cacagtggcg gcccgcacgc ggccctgttc
     1681 caccaggggc cccaggcgct cgcggatggc gctgaggccg cgctcggcgc cctcgcgggc
     1741 cccggcctgg tacactgcca ggcgcttctg caggtcatcg gcatcgcgga ggagccgctt
     1801 acgcagcttg cgcaggtggg aggcgaggcg cacccgcagc tcctcggtgc tctggccgag
     1861 catggcctgc acctcgccgc ggtactgcac caggcggccg cgcacgtcct ccatgtccgc
     1921 gcccagccgg gcctgcgccg cctgcagctc cttggacagc cgtgcccgcg tctcctccgc
     1981 caccggggtc agttgttcct ccagttccga tttgtaggcc ttcaactcct tcatggtctc
     2041 gtccatcagc gccctcagtt cctgggtgac ctgggagctg agcagctcct cctgcacctg
     2101 ctcagacagt gtctgcaccc agcgcaggta atcccaaaag cgacccagtg ccagttccca
     2161 gcgctggccg ctctgccact cggtctgctg gcgcagctcg ggctccggct ctgtctccac
     2221 cgcttgctcc accttacctc taactaaaac attggaggag tggaaagaat tctgtaaagg
     2281 taatattacc acgaaaaggg cgtacaggac cacggagagt ggtaccagca gcttgccagc
     2341 gcttagtctg ttgaaaaaca ttttgttact cgagtatagt tttttctcct tgacgttaaa
     2401 gtatagaggt atattaacaa ttttttgttg atacttttat gacatttgaa taagaagtaa
     2461 tacaaactga aaatgttgaa agtattagtt aaagtggtta tgcagctttt ccatttatat
     2521 atctgttaat agatcaaaaa tcatcgcttc gctgattaat taccccagaa ataaggctaa
     2581 aaaactaatc gcattatcat cctatggttg ttaatttgat tcgttaattt gaaggtttgt
     2641 ggggccaggt tactgccaat ttttcctctt cataaccata aaagctagta ttgtagaatc
     2701 tttattgttc ggagcagtgc ggcattaggc acatctgcgt ttcaggaacg cgaccggtga
     2761 agacgaggac gcacggtcta gacgcccacg cacgcccacg cccgcccacg cacgcccacg
     2821 cccgcccacg cacgcccacg cggccgcact tcaatatagc aatgagcagt taagcgtatt
     2881 actgaaagtt ccaaagagaa ggttttttta ggctaagata atggggctct ttacatttcc
     2941 acaacatata agtaagatta gatatggata tgtatatgga tatgtatatg gtggtaatgc
     3001 catgtaatat gattattaaa cttctttgcg tccatccaaa aaaaaagtaa gaatttttga
     3061 aaattcaata taagggccgg taccgttcgt ttcccgatac atgcgaatgc acaagctaat
     3121 atagccttat cagatgattc accggtttga tagtcattac taaatgcttg caaggattgc
     3181 gatctggtca aaaccaatat agtcactgat ttctaaaccg aactagatat gatatttact
     3241 tctttctgtt cagacttatg tatgcaactg tacatctact aagtctaagc taatagttcc
     3301 ctgacagctt tcgctactgc atcaccgacc tcagtagtag agttggtacc ctttagatcc
     3361 gctgttctga taccagaatc caacactttc ttgactgctt gttctataat ttcaccttct
     3421 tcgaacaggt ccagagacaa cttcaacatc atagcagcag ataggattgt tgctacgggg
     3481 ttaacctttc cctttggtaa gtctggagca gagccatgac atggctcata taaaccaaat
     3541 gccgtattct tatctggaag agatgccaag gaggcagatg gaagtaatcc taacgaacct
     3601 gggatcacgg aggcttcatc ggagataata tcaccaaaca tattgtttgt gataataata
     3661 ccatttagat gagttgggtt cttaaccaag atcatagcag cggaatcaat caattgatgt
     3721 tgaacagtca attgtgggaa ttcattctta atcgtctctt ccactgtttt tctccataat
     3781 cttgatgatg ctagaacgtt ggctttatct agagaccaaa taggtaaagg tgggttgtgt
     3841 tgcagagcca tgaaagcagc cattctagtg attctttgaa cttcgggcac agaatacttt
     3901 tcgctatccc aggcgacacc atcaccttca tcttcttttc tctccccgaa ataaatacca
     3961 cccactagtt ctctaacaac gacgaaatct gtacctcttg caatttcagg cttcaatggc
     4021 gatagatcta gtaaggaatc agatgcaaaa ttacatggtc ttagattagc atacaattgc
     4081 aactccttac ggattttcaa aagaccttgt tctggtctga cagcgccagt accccatttt
     4141 ggaccaccaa cagcacccag aagcacggca tcagctttct tagaggcctc caaagcttcg
     4201 tctggtaatg gaacaccagt ggcatcaatt gcagcaccac cgatcaaatg atgatcaaac
     4261 ttgaaattga ccttgtctgg acgacattcc tgaatagcat ttaaaacttt aatggcctct
     4321 tcagtgattt cttgaccaac atggtcacct ggtagaacta caattgtctt ggtcacagcc
     4381 atgattattc tttatttatt agttgtgtat agtgtattct ctttcaagtt tgttctttaa
     4441 ggattatggt actttaagta ctatatgaac aatgggagga tacaagaaaa ctgtaagaca
     4501 agcttcaaca caatggttta tatatacatt gctttcaatt ggaaatacat cctcgattca
     4561 aaagttagtc attgacttta ctgtgaccgg ttgaacaaat tactacgaaa gttgaatcat
     4621 tttagcttgc taggggcttt tgcccggtac cggtgccggg ctcttttgag cgatacaggt
     4681 cacatgcgca acgatcataa ccagtttcaa gatagcgatg aatacagagc acacatatac
     4741 gggaaacaca gaaatttgat aactacaatg ggatgaactg aaaacttaaa ctttcatatt
     4801 ttcgacagtg aaactattaa cttttgattt tcacaaaata tttcttgttg aaaccttata
     4861 ttaaaccgct atctattggt aacttagggg cagacattag aatggtatat ccttgaaata
     4921 tatatatata tattgctgaa atgtaaaagg taagaaaagt tagaaagtaa gacgattgct
     4981 aaccacctat tggaaaaaac aataggtcct taaataatat tgtcaacttc aagtattgtg
     5041 atgcaagcat ttagtcatga acgcttctct attctatatg aaaagccggt tccggcgctc
     5101 tcacctttcc tttttctccc aatttttcag ttgaaaaagg tatatgcgtc aggcgacctc
     5161 tgaaattaac aaaaaatttc cagtcatcga atttgattct gtgcgatagc gcccctgtgt
     5221 gttctcgtta tgttgaggaa aaaaataatg gttgctaaga gattcgaact cttgcatctt
     5281 acgatacctg agtattccca cagttaactg cggtcaagat atttcttgaa tcaggcgcct
     5341 tagaccgctc ggccaaacaa ccaattactt gttgagaaat agagtataat tatcctataa
     5401 atataacgtt tttgaacaca catgaacaag gaagtacagg acaattgatt ttgaagagaa
     5461 tgtggatttt gatgtaatcg ttgggattcc atttttaata aggcaataat attaggtatg
     5521 tagatatact agaagttctc ctcggccagc gtggccgttt aaacatttaa atggccggcc
     5581 cagcttttgt tccctttagt gagggttaat tccgagcttg gcgtaatcat ggtcatagct
     5641 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     5701 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     5761 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     5821 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     5881 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5941 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     6001 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6061 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6121 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6181 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     6241 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6301 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6361 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6421 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     6481 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6541 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     6601 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6661 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6721 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6781 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6841 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6901 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     6961 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     7021 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     7081 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     7141 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     7201 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     7261 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     7321 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     7381 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     7441 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     7501 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     7561 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     7621 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     7681 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     7741 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcc tcggcgcgcc
     7801 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     7861 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     7921 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     7981 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     8041 ctgagagtgc accataccac cttttcaatt catcattttt tttttattct tttttttgat
     8101 ttcggtttcc ttgaaatttt tttgattcgg taatctccga acagaaggaa gaacgaagga
     8161 aggagcacag acttagattg gtatatatac gcatattggg tcagcagtac agaaccgtcg
     8221 acgaagttgt aagtggtgga tcagatatca tcattgttgg cagaggactt ttcgccaagg
     8281 gtagagatcc taatactaaa ctcacaaatt agagcttcaa tttaattata tcagttatta
     8341 ccctatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggaaatt
     8401 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     8461 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     8521 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     8581 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     8641 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     8701 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     8761 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     8821 gccgcgctta atgcgccgct acagggcgcg tccattcgcc attcaggctg cgcaactgtt
     8881 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     8941 ctgcaaggcg attaagttgg gtaacgccag ggt
//
