LOCUS       sequence-364549-        5979 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses ApoE4 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-ApoE4
FEATURES             Location/Qualifiers
     source          1..5979
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     287..307
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    331..953
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     735..754
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     promoter        complement(2199..2402)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2228..2248)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2403..2782
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(2943..2960)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2943..2959)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2952..2974)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(3100..3555)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3237..3258)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3449..3468
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3582..3686
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3687..3755,3756..4547)
                     /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(3905..3924)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4718..5306
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5207..5226
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5460..5477
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5594..5615
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5630..5635,5636..5653,5654..5660)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5668..5684
                     /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     5673..5695
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5692..5708
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5692..5708
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5722..5741
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5722..5740
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     intron          5817..5913
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     promoter        complement(join(5952..5979,1..302))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(5966..5979,1..122)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 cctcctcact acttctggaa tagctcagag gccgaggcgg cctcggcctc tgcataaata
       61 aaaaaaatta gtcagccatg gggcggagaa tgggcggaac tgggcggagt taggggcggg
      121 atgggcggag ttaggggcgg gactatggtt gctgactaat tgagatgcat gctttgcata
      181 cttctgcctg ctggggagcc tggggacttt ccacacctgg ttgctgacta attgagatgc
      241 atgctttgca tacttctgcc tgctggggag cctggggact ttccacaccc taactgacac
      301 acattccaca gccaagctgg ccgcgtaccc aattcaacag gcatctactg agtggaccca
      361 acgcatgaga ggacagtgcc aagcaagcaa ctcaaatgtc ccaccggttg ggcatggcca
      421 ggtagcctat gctgtgtctg gacgtcctcc tgctggtata gttattttaa aatcagaagg
      481 acagggaagg gagcagtggt tcacgcctgt aatcccagca atttgggagg ccaaggtggg
      541 tagatcacct gagattagga gttggagacc agcctggcca atatggtgaa accccgtctc
      601 taccaaaaaa acaaaaatta gctgagcctg gtcatgcatg cctggaatcc caacaactcg
      661 ggaggctgag gcaggagaat cgcttgaacc caggaggcgg agattgcagt gagccaagat
      721 tgtgccactg cactccagct tggttcccaa tagaccccgc aggccctaca ggttgtcttc
      781 ccaacttgcc ccttgctcca taccaccccc ctccacccca taatattata gaaggacacc
      841 tagtcagaca aaatgatgca acttaatttt attaggacaa ggctggtggg cactggagtg
      901 gcaacttcca gggccaggag aggcactggg gaggggtcac agggatgcca cccgggatcc
      961 tctagaggat ccccgggaat ctttattaaa ctagggtcca ccccaggagg acggctgggg
     1021 cggggacagg gtctcccgct gcaggctgcg cggaggcagg aggcacgggg tggcgtgggg
     1081 tcgcatggct gcaggcttcg gcgttcagtg attgtcgctg ggcacagggg cggcgctggt
     1141 gcccacggca gcctgcacct tctccaccag cccggcccac tggcgctgca tgtcttccac
     1201 caggggctcg aaccagctct tgaggcgggc ctggaaggcc tcggcctgca ggcgtatctg
     1261 ctgggcctgc tcctccagct tggcgcgcac ctccgccacc tgctccttca cctcgtccag
     1321 gcggtcgcgg gtccggctgc ccatctcctc catccgcgcg cgcagccgct cgccccaggc
     1381 ctgggcccgc tcctgtagcg gctggccggc cagggagccc acagtggcgg cccgcacgcg
     1441 gccctgttcc accaggggcc ccaggcgctc gcggatggcg ctgaggccgc gctcggcgcc
     1501 ctcgcgggcc ccggcctggt acactgccag gcgcttctgc aggtcatcgg catcgcggag
     1561 gagccgctta cgcagcttgc gcaggtggga ggcgaggcgc acccgcagct cctcggtgct
     1621 ctggccgagc atggcctgca cctcgccgcg gtactgcacc aggcggccgc gcacgtcctc
     1681 catgtccgcg cccagccggg cctgcgccgc ctgcagctcc ttggacagcc gtgcccgcgt
     1741 ctcctccgcc accggggtca gttgttcctc cagttccgat ttgtaggcct tcaactcctt
     1801 catggtctcg tccatcagcg ccctcagttc ctgggtgacc tgggagctga gcagctcctc
     1861 ctgcacctgc tcagacagtg tctgcaccca gcgcaggtaa tcccaaaagc gacccagtgc
     1921 cagttcccag cgctggccgc tctgccactc ggtctgctgg cgcagctcgg gctccggctc
     1981 tgtctccacc gcttgctcca ccttggcctg gcatcctgcc aggaatgtga ccagcaacgc
     2041 agcccacaga accttcatct tcctgcctgt gattggcggg atcccgcggg atcccgccag
     2101 tcggctcctg gggaaggccc cgggtaccag atctgaattc gatggaagta tttgaaagaa
     2161 aattaaaaat aaaaacaatt ctgacggttc actaaacgag ctctgcttat atagacctcc
     2221 caccgtacac gcctaccgcc catttgcgtc aacggggcgg ggttattacg acattttgga
     2281 aagtcccgtt gattttggtg ccaaaacaaa ctcccattga cgtcaatggg gtggagactt
     2341 ggaaatcccc gtgagtcaaa ccgctatcca cgcccattgg tgtactgcca aaaccgcatc
     2401 accatggtaa tagcgatgac taatacgtag atgtactgcc aagtaggaaa gtcccgtaag
     2461 gtcatgtact gggcataatg ccaggcgggc catttaccgt cattgacgtc aatagggggc
     2521 ggacttggca tatgatacac ttgatgtact gccaagtggg cagtttaccg taaatactcc
     2581 acccattgac gtcaatggaa agtccctatt ggcgttacta tgggaacata cgtcattatt
     2641 gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg ggccatttac cgtaagttat
     2701 gtaacgcgga actccatata tgggctatga actaatgacc ccgtaattga ttactattaa
     2761 taactagtca ataatcaatg tcaacatggc ggtcatattg gacatgagcc aatataaatg
     2821 tacatattat gatatagata caacgtatgc aatggccaat agccaatatt gatttatgct
     2881 atataaccaa tgactaatat ggctaattgc caatattgat tcaatgtata gatcagcttg
     2941 gcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     3001 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     3061 cgcccttccc aacagttgcg cagcctgaat ggcgaatgga aattgtaagc gttaatattt
     3121 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     3181 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     3241 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     3301 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     3361 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     3421 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     3481 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     3541 cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     3601 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     3661 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     3721 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     3781 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     3841 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     3901 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     3961 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     4021 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     4081 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     4141 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     4201 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     4261 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     4321 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     4381 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     4441 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     4501 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     4561 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     4621 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     4681 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     4741 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     4801 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     4861 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     4921 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     4981 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     5041 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     5101 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     5161 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     5221 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     5281 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     5341 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     5401 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     5461 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     5521 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     5581 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
     5641 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
     5701 gctatgacat gattacgaat ttaatacgac tcactatagg gaattcgagc tcggtacgcg
     5761 gccctagagt cgacctgcag cagcttttag agcagaagta acacttccgt acaggcctag
     5821 aagtaaaggc aacatccact gaggagcagt tctttgattt gcaccaccac cggatccggg
     5881 acctgaaata aaagacaaaa agactaaact taccagttaa ctttctggtt tttcagttcc
     5941 tcgaggagct ttttgcaaaa gcctaggcct ccaaaaaag
//
