LOCUS       sequence-334592-        5979 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses ApoE3 in Mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5979)
  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 5979)
  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: pCMV4-ApoE3
FEATURES             Location/Qualifiers
     source          1..5979
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     126..148
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     140..157
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     141..157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        318..697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     852..872
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    2147..2769
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2346..2365)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2793..2813)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2798..3127
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2978..3113
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          3166..3262
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     primer_bind     complement(3338..3357)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3339..3357)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3371..3387)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3371..3387)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3384..3406)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3395..3411
                     /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(3419..3425,3426..3443,3444..3449))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3464..3485
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3602..3619)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3773..4361)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3853..3872)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4532..5323,5324..5392))
                     /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     5155..5174
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5393..5497)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5524..5952
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5611..5630)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5821..5842
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attgttttta tttttaattt tctttcaaat acttccatcg
      961 aattcagatc tggtacccgg ggccttcccc aggagccgac tggcgggatc ccgcgggatc
     1021 ccgccaatca caggcaggaa gatgaaggtt ctgtgggctg cgttgctggt cacattcctg
     1081 gcaggatgcc aggccaaggt ggagcaagcg gtggagacag agccggagcc cgagctgcgc
     1141 cagcagaccg agtggcagag cggccagcgc tgggaactgg cactgggtcg cttttgggat
     1201 tacctgcgct gggtgcagac actgtctgag caggtgcagg aggagctgct cagctcccag
     1261 gtcacccagg aactgagggc gctgatggac gagaccatga aggagttgaa ggcctacaaa
     1321 tcggaactgg aggaacaact gaccccggtg gcggaggaga cgcgggcacg gctgtccaag
     1381 gagctgcagg cggcgcaggc ccggctgggc gcggacatgg aggacgtgtg cggccgcctg
     1441 gtgcagtacc gcggcgaggt gcaggccatg ctcggccaga gcaccgagga gctgcgggtg
     1501 cgcctcgcct cccacctgcg caagctgcgt aagcggctcc tccgcgatgc cgatgacctg
     1561 cagaagcgcc tggcagtgta ccaggccggg gcccgcgagg gcgccgagcg cggcctcagc
     1621 gccatccgcg agcgcctggg gcccctggtg gaacagggcc gcgtgcgggc cgccactgtg
     1681 ggctccctgg ccggccagcc gctacaggag cgggcccagg cctggggcga gcggctgcgc
     1741 gcgcggatgg aggagatggg cagccggacc cgcgaccgcc tggacgaggt gaaggagcag
     1801 gtggcggagg tgcgcgccaa gctggaggag caggcccagc agatacgcct gcaggccgag
     1861 gccttccagg cccgcctcaa gagctggttc gagcccctgg tggaagacat gcagcgccag
     1921 tgggccgggc tggtggagaa ggtgcaggct gccgtgggca ccagcgccgc ccctgtgccc
     1981 agcgacaatc actgaacgcc gaagcctgca gccatgcgac cccacgccac cccgtgcctc
     2041 ctgcctccgc gcagcctgca gcgggagacc ctgtccccgc cccagccgtc ctcctggggt
     2101 ggaccctagt ttaataaaga ttcccgggga tcctctagag gatcccgggt ggcatccctg
     2161 tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct
     2221 tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat
     2281 ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg
     2341 cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc
     2401 cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg gattccaggc
     2461 atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt
     2521 ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt
     2581 gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt taaaataact
     2641 ataccagcag gaggacgtcc agacacagca taggctacct ggccatgccc aaccggtggg
     2701 acatttgagt tgcttgcttg gcactgtcct ctcatgcgtt gggtccactc agtagatgcc
     2761 tgttgaattg ggtacgcggc cagcttggct gtggaatgtg tgtcagttag ggtgtggaaa
     2821 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     2881 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     2941 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     3001 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     3061 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     3121 ttgcaaaaag ctcctcgagg aactgaaaaa ccagaaagtt aactggtaag tttagtcttt
     3181 ttgtctttta tttcaggtcc cggatccggt ggtggtgcaa atcaaagaac tgctcctcag
     3241 tggatgttgc ctttacttct aggcctgtac ggaagtgtta cttctgctct aaaagctgct
     3301 gcaggtcgac tctagggccg cgtaccgagc tcgaattccc tatagtgagt cgtattaaat
     3361 tcgtaatcat gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     3421 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     3481 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     3541 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     3601 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     3661 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     3721 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     3781 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     3841 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     3901 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     3961 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     4021 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4081 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     4141 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     4201 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     4261 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     4321 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     4381 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     4441 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     4501 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     4561 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     4621 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     4681 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     4741 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     4801 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     4861 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     4921 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     4981 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     5041 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     5101 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     5161 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     5221 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     5281 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     5341 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     5401 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     5461 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     5521 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     5581 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     5641 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     5701 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     5761 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     5821 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     5881 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     5941 ttaacgcgaa ttttaacaaa atattaacgc ttacaattt
//
