LOCUS       sequence-319719-        7935 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 7935)
  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 7935)
  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_107
FEATURES             Location/Qualifiers
     source          1..7935
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..344
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      1163..1360
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     promoter        1515..1956
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    1515..1632
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     1927..1950
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     promoter        complement(3929..3947)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3968..3984)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3968..3984)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3981..4003)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3992..4008
                     /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(4016..4022,4023..4040,4041..4046))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4061..4082
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4199..4216)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4370..4958)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4450..4469)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5129..5920,5921..5989))
                     /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     5752..5771
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5990..6094)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6179..6197
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6235..6257)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6357..6376
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(6732..7187)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6869..6890)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7081..7100
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7313..7335
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7327..7344
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7328..7344
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7351..7370
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7351..7369
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gacatggagg cccagaatac cctccttgac agtcttgacg tgcgcagctc aggggcatga
       61 tgtgactgtc gcccgtacat ttagcccata catccccatg tataatcatt tgcatccata
      121 cattttgatg gccgcacggc gcgaagcaaa aattacggct cctcgctgcg gacctgcgag
      181 cagggaaacg ctcccctcac agacgcgttg aattgtcccc acgccgcgcc cctgtagaga
      241 aatataaaag gttaggattt gccactgagg ttcttctttc atatacttcc ttttaaaatc
      301 ttgctaggat acagttctca catcacatcc gaacataaac aaccatgaca gtcaacacta
      361 agacctatag tgagagagca gaaactcatg cctcaccagt agcacaacga ttatttcgat
      421 taatggaact gaagaaaacc aatttatgtg catcaattga tgttgatacc actaaggaat
      481 tccttgaatt aattgataaa ttgggtcctt atgtatgctt aatcaagact catattgata
      541 taatcaatga tttttcctat gaatccacta ttgaaccatt attagaactt tcacgtaaac
      601 atcaatttat gatttttgaa gatagaaaat ttgctgatat tggtaatacc gtgaagaaac
      661 aatatattgg tggagtttat aaaattagta gttgggcaga tattactaat gctcatggtg
      721 tcactgggaa tggagtagtt gaaggattaa aacagggagc taaagaaacc accaccaacc
      781 aagagccaag agggttattg atgttagctg aattatcatc agtgggatca ttagcatatg
      841 gagaatattc tcaaaaaact gttgaaattg ctaaatccga taaggaattt gttattggat
      901 ttattgccca acgtgatatg ggtggacaag aagaaggatt tgattggctt attatgacac
      961 ctggagttgg attagatgat aaaggtgatg gattaggaca acaatataga actgttgatg
     1021 aagttgttag cactggaact gatattatca ttgttggtag aggattgttt ggtaaaggaa
     1081 gagatccaga tattgaaggt aaaaggtata gagatgctgg ttggaatgct tatttgaaaa
     1141 agactggcca attataatca gtactgacaa taaaaagatt cttgttttca agaacttgtc
     1201 atttgtatag tttttttata ttgtagttgt tctattttaa tcaaatgtta gcgtgattta
     1261 tatttttttt cgcctcgaca tcatctgccc agatgcgaag ttaagtgcgc agaaagtaat
     1321 atcatgcgtc aatcgtatgt gaatgctggt cgctatactg ctgtcgattc gatactaacg
     1381 ccgccatcca gtgtcgaaaa cgagctcgaa ttcatcgagt ggattcggct ttgggtaccg
     1441 ggcccaaata cctcgaggga gcaaggcagg tggtcgacgg tatcgataag ctaacaaaag
     1501 ctggagctct agtacggatt agaagccgcc gagcgggtga cagccctccg aaggaagact
     1561 ctcctccgtg cgtcctcgtc ttcaccggtc gcgttcctga aacgcagatg tgcctcgcgc
     1621 cgcactgctc cgaacaataa agattctaca atactagctt ttatggttat gaagaggaaa
     1681 aattggcagt aacctggccc cacaaacctt caaatgaacg aatcaaatta acaaccatag
     1741 gatgataatg cgattagttt tttagcctta tttctggggt aattaatcag cgaagcgatg
     1801 atttttgatc tattaacaga tatataaatg caaaaactgc ataaccactt taactaatac
     1861 tttcaacatt ttcggtttgt attacttctt attcaaatgt aataaaagta tcaacaaaaa
     1921 attgttaata tacctctata ctttaacgtc aaggagaaaa aaccaaaaca actaattcga
     1981 aatgtttttc aacagactaa gcgctggcaa gctgctggta ccactctccg tggtcctgta
     2041 cgcccttttc gtggtaatat tacctttaca gaattctttc cactcctcca atgttttagt
     2101 tagaggtaag gtggagcaag cggtggagac agagccggag cccgagctgc gccagcagac
     2161 cgagtggcag agcggccagc gctgggaact ggcactgggt cgcttttggg attacctgcg
     2221 ctgggtgcag acactgtctg agcaggtgca ggaggagctg ctcagctccc aggtcaccca
     2281 ggaactgagg gcgctgatgg acgagaccat gaaggagttg aaggcctaca aatcggaact
     2341 ggaggaacaa ctgaccccgg tggcggagga gacgcgggca cggctgtcca aggagctgca
     2401 ggcggcgcag gcccggctgg gcgcggacat ggaggacgtg cgcggccgcc tggtgcagta
     2461 ccgcggcgag gtgcaggcca tgctcggcca gagcaccgag gagctgcggg tgcgcctcgc
     2521 ctcccacctg cgcaagctgc gtaagcggct cctccgcgat gccgatgacc tgcagaagcg
     2581 cctggcagtg taccaggccg gggcccgcga gggcgccgag cgcggcctca gcgccatccg
     2641 cgagcgcctg gggcccctgg tggaacaggg ccgcgtgcgg gccgccactg tgggctccct
     2701 ggccggccag ccgctacagg agcgggccca ggcctggggc gagcggctgc gcgcgcggat
     2761 ggaggagatg ggcagccgga cccgcgaccg cctggacgag gtgaaggagc aggtggcgga
     2821 ggtgcgcgcc aagctggagg agcaggccca gcagatacgc ctgcaggccg aggccttcca
     2881 ggcccgcctc aagagctggt tcgagcccct ggtggaagac atgcagcgcc agtgggccgg
     2941 gctggtggag aaggtgcagg ctgccgtggg caccagcgcc gcccctgtgc ccagcgacaa
     3001 tcactgaggc cgccaccgcg gtggagctct aagcaaatag ctaaattata tacgaattaa
     3061 tattatgatt aagtgtttac gtgagtgcga tatttttatt actatcttat acagttgtat
     3121 atactctata aaatgagttg tctattaatt aacgcgatga atgctttctg ggtttacctc
     3181 tccaacaact ctagtttact tctcaataca ttcaattgta tttgatttgt caatacttca
     3241 tcattaatca attctatagt tttgtttttc tcgtttattt ccaaatttaa tgcatcaatt
     3301 ttattattca atttgtcgtt gattttggtt aatgatttta tggtttgatc tctggcattg
     3361 attgtttgtg ttagtttttc attattgata attaaattat ttaagttagt tatcaactcg
     3421 gtgttttcaa gtttcaagtt ttcaatttct ttagagttta ttagatttgt caaagtttct
     3481 gaattgcttg attggtccgg cccaggtatt gttagcggtt tgaagcaggc ggcagaagaa
     3541 gtaacaaagg aacctagagg ccttttgatg ttagcagaat tgtcatgcaa gggctcccta
     3601 tctactggag aatatactaa gggtactgtt gacattgcga agagcgacaa agattttgtt
     3661 atcggcttta ttgctcaaag agacatgggt ggaagagatg aaggttacga ttggttgatt
     3721 atgacacccg gtgtgggttt agatgacaag ggagacgcat tgggtcaaca gtatagaacc
     3781 gtggatgatg tggtctctac aggatctgac attattattg ttggaagagg actatttgca
     3841 aagggaaggg atgctaaggt agagggtgaa cgttacagaa aagcaggctg gggtttaaac
     3901 atttaaatgg ccggcccagc ttttgttccc tttagtgagg gttaattccg agcttggcgt
     3961 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     4021 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     4081 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     4141 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     4201 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4261 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4321 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4381 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4441 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4501 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4561 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4621 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4681 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4741 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4801 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4861 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4921 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4981 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     5041 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     5101 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     5161 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     5221 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     5281 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     5341 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     5401 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     5461 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     5521 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     5581 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     5641 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     5701 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     5761 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     5821 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     5881 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     5941 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     6001 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     6061 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     6121 acgtcctcgg cgcgccgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     6181 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     6241 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     6301 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     6361 atcagagcag attgtactga gagtgcacca taccaccttt tcaattcatc attttttttt
     6421 tattcttttt tttgatttcg gtttccttga aatttttttg attcggtaat ctccgaacag
     6481 aaggaagaac gaaggaagga gcacagactt agattggtat atatacgcat attgggtcag
     6541 cagtacagaa ccgtcgacga agttgtaagt ggtggatcag atatcatcat tgttggcaga
     6601 ggacttttcg ccaagggtag agatcctaat actaaactca caaattagag cttcaattta
     6661 attatatcag ttattaccct atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     6721 accgcatcag gaaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     6781 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     6841 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     6901 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     6961 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     7021 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     7081 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     7141 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcca ttcgccattc
     7201 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     7261 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
     7321 cgacgttgta aaacgacggc cagtgaattg taatacgact cactataggg ggccagcgtg
     7381 gccgtttaaa cagaacgaag gaaggagcac agacttagat tggtatatat acgcatatgt
     7441 agtgttgaag aaacatgaaa ttgcccagta ttcttaaccc aactgcacag aacaaaaacc
     7501 tgcaggaaac gaagataaat catgtcgaaa gctacatata aggaacgtgc tgctactcat
     7561 cctagtcctg ttgctgccaa gctatttaat atcatgcacg aaaagcaaac aaacttgtgt
     7621 gcttcattgg atgttcgtac caccaaggaa ttactggagt tagttgaagc attaggtccc
     7681 aaaatttgtt tactaaaaac acatgtggat atcttgactg atttttccat ggagggcaca
     7741 gttaagccgc taaaggcatt atccgccaag tacaattttt tactcttcga agacagaaaa
     7801 tttgctgaca ttggtaatac agtcaaattg cagtactctg cgggtgtata cagaatagca
     7861 gaatgggcag acattacgaa tgcacacggt gtggtggttt agcttgcctc gtccccgccg
     7921 ggtcacccgg ccagc
//
