LOCUS       sequence-388019-        6519 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses ApoE4 in Mammalian cell.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6519)
  AUTHORS   Hudry E, Dashkoff J, Roe AD, Takeda S, Koffie RM, Hashimoto T, 
            Scheel M, Spires-Jones T, Arbel-Ornath M, Betensky R, Davidson BL, 
            Hyman BT
  TITLE     Gene transfer of human Apoe isoforms results in differential 
            modulation of amyloid deposition and neurotoxicity in mouse brain.
  JOURNAL   Sci Transl Med. 2013 Nov 20;5(212):212ra161. doi: 
            10.1126/scitranslmed.3007000.
  PUBMED    24259049
REFERENCE   2  (bases 1 to 6519)
  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: "2013-11-20"; volume: "5(212):212ra161. doi"; pages: " 
            10.1126/scitranslmed.3007000"
COMMENT     Sequence Label: pcDNA3-ApoE4
FEATURES             Location/Qualifiers
     source          1..6519
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(58..77)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        249..628
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        629..832
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     783..803
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     877..896
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        877..895
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2086..2104)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2087..2104)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2124..2141)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2130..2354
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2400..2828
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2487..2506)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2697..2718
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2837..2857)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2842..3171
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3022..3157
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3084..3103
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3238..4032
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3292..3311)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3902..3921
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4206..4327
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4243..4262)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4297..4316
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4376..4392)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4376..4392)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4389..4411)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4400..4416
                     /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(4424..4430,4431..4448,4449..4454))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4469..4490
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4607..4624)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4778..5366)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4858..4877)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5537..6328,6329..6397))
                     /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     6160..6179
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6398..6502)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac
       61 aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt
      121 cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg
      181 catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata
      241 tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
      301 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      361 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      421 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      481 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      541 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      601 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
      661 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
      721 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
      781 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctctctggct
      841 aactagagaa cccactgctt actggcttat cgaaattaat acgactcact atagggagac
      901 ccaagcttgg tacccggggc cttccccagg agccgactgg cgggatcccg cgggatcccg
      961 ccaatcacag gcaggaagat gaaggttctg tgggctgcgt tgctggtcac attcctggca
     1021 ggatgccagg ccaaggtgga gcaagcggtg gagacagagc cggagcccga gctgcgccag
     1081 cagaccgagt ggcagagcgg ccagcgctgg gaactggcac tgggtcgctt ttgggattac
     1141 ctgcgctggg tgcagacact gtctgagcag gtgcaggagg agctgctcag ctcccaggtc
     1201 acccaggaac tgagggcgct gatggacgag accatgaagg agttgaaggc ctacaaatcg
     1261 gaactggagg aacaactgac cccggtggcg gaggagacgc gggcacggct gtccaaggag
     1321 ctgcaggcgg cgcaggcccg gctgggcgcg gacatggagg acgtgcgcgg ccgcctggtg
     1381 cagtaccgcg gcgaggtgca ggccatgctc ggccagagca ccgaggagct gcgggtgcgc
     1441 ctcgcctccc acctgcgcaa gctgcgtaag cggctcctcc gcgatgccga tgacctgcag
     1501 aagcgcctgg cagtgtacca ggccggggcc cgcgagggcg ccgagcgcgg cctcagcgcc
     1561 atccgcgagc gcctggggcc cctggtggaa cagggccgcg tgcgggccgc cactgtgggc
     1621 tccctggccg gccagccgct acaggagcgg gcccaggcct ggggcgagcg gctgcgcgcg
     1681 cggatggagg agatgggcag ccggacccgc gaccgcctgg acgaggtgaa ggagcaggtg
     1741 gcggaggtgc gcgccaagct ggaggagcag gcccagcaga tacgcctgca ggccgaggcc
     1801 ttccaggccc gcctcaagag ctggttcgag cccctggtgg aagacatgca gcgccagtgg
     1861 gccgggctgg tggagaaggt gcaggctgcc gtgggcacca gcgccgcccc tgtgcccagc
     1921 gacaatcact gaacgccgaa gcctgcagcc atgcgacccc acgccacccc gtgcctcctg
     1981 cctccgcgca gcctgcagcg ggagaccctg tccccgcccc agccgtcctc ctggggtgga
     2041 ccctagttta ataaagattc ccggggatcc tctagagggc cctattctat agtgtcacct
     2101 aaatgctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     2161 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     2221 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     2281 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     2341 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     2401 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     2461 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     2521 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     2581 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     2641 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     2701 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     2761 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     2821 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     2881 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     2941 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3001 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3061 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     3121 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     3181 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     3241 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     3301 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     3361 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     3421 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     3481 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     3541 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     3601 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     3661 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     3721 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     3781 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     3841 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     3901 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     3961 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     4021 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     4081 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     4141 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     4201 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     4261 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     4321 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     4381 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     4441 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     4501 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     4561 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     4621 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     4681 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4741 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4801 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4861 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4921 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4981 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     5041 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     5101 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     5161 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     5221 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     5281 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5341 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5401 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5461 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     5521 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5581 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     5641 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     5701 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     5761 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     5821 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     5881 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     5941 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     6001 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     6061 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     6121 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     6181 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     6241 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     6301 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     6361 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     6421 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6481 ataaacaaat aggggttccg cgcacatttc cccgaaaag
//
