LOCUS       sequence-288270-        6271 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  high transduction efficiency AAV-mediated synapsin 
            promoter-dependent expression of AICD (last 50 amino acids of APP) 
            and IRES-mediated co-expression of hrGFP .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6271)
  AUTHORS   Pousinha PA, Mouska X, Raymond EF, Gwizdek C, Dhib G, Poupon G, 
            Zaragosi LE, Giudici C, Bethus I, Pacary E, Willem M, Marie H
  TITLE     Physiological and pathophysiological control of synaptic GluN2B-NMDA
            receptors by the C-terminal domain of amyloid precursor protein.
  JOURNAL   Elife. 2017 Jul 6;6. doi: 10.7554/eLife.25659.
  PUBMED    28682239
REFERENCE   2  (bases 1 to 6271)
  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: "Elife."; 
            date: "2017-07-6"; volume: "6. doi"; pages: " 10.7554/eLife.25659"
COMMENT     Sequence Label: pAAV-syn-AICD-IRES-hrGFP
FEATURES             Location/Qualifiers
     source          1..6271
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   22..151
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        362..809
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      822..831
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             990..1061
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     misc_feature    join(1097..1668,1669..1671,1672..1680)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1264..1281)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1491..1510
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1681..2400
                     /codon_start=1
                     /product="humanized Renilla green fluorescent protein"
                     /label=hrGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKQILKNTGLQEIMSFKVNLEGVVNNHVFTMEGCGKGNILFGN
                     QLVQIRVTKGAPLPFAFDILSPAFQYGNRTFTKYPEDISDFFIQSFPAGFVYERTLRYE
                     DGGLVEIRSDINLIEEMFVYRVEYKGRNFPNDGPVMKKTITGLQPSFEVVYMNDGVLVG
                     QVILVYRLNSGKFYSCHMRTLMKSKGVVKDFPEYHFIQHRLEKTYVEDGGFVEQHETAI
                     AQLTSLGKPLGSLHEWV"
     misc_feature    2425..3013
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2478..2498)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2896..2907)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3033..3509
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3232..3251)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3555..3695
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3555..3684
                     /label=AAV2 ITR
     rep_origin      3770..4225
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3857..3876)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4067..4088
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4242..4261)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4361..4383
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4421..4439)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4507..4611
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4612..4680,4681..5472)
                     /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(4830..4849)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5643..6231
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6132..6151
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gctggccttt tgctcacatg tcctgcaggc agctgcgcgc tcgctcgctc actgaggccg
       61 cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag
      121 ggagtggcca actccatcac taggggttcc tgcggccgca cgcgttgcaa agatggataa
      181 agttttaaac agagaggaat ctttgcagct aatggacctt ctaggtcttg aaaggagtgg
      241 gaattggctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt
      301 tggggggagg ggtcggcagc aaatggttaa ttaatctaga ctgcagaggg ccctgcgtat
      361 gagtgcaagt gggttttagg accaggatga ggcggggtgg gggtgcctac ctgacgaccg
      421 accccgaccc actggacaag cacccaaccc ccattcccca aattgcgcat cccctatcag
      481 agagggggag gggaaacagg atgcggcgag gcgcgtgcgc actgccagct tcagcaccgc
      541 ggacagtgcc ttcgcccccg cctggcggcg cgcgccaccg ccgcctcagc actgaaggcg
      601 cgctgacgtc actcgccggt cccccgcaaa ctccccttcc cggccacctt ggtcgcgtcc
      661 gcgccgccgc cggcccagcc ggaccgcacc acgcgaggcg cgagataggg gggcacgggc
      721 gcgaccatct gcgctgcggc gccggcgact cagcgctgcc tcagtctgcg gtgggcagcg
      781 gaggagtcgt gtcgtgcctg agagcgcagt cgagaggatc cgccaccatg gtgatgctga
      841 agaagaaaca gtacacatcc attcatcatg gtgtggtgga ggttgacgcc gctgtcaccc
      901 cagaggagcg ccacctgtcc aagatgcagc agaacggcta cgaaaatcca acctacaagt
      961 tctttgagca gatgcagaac tgactcgagg actacaagga tgacgatgac aaggattaca
     1021 aagacgacga tgataaggac tataaggatg atgacgacaa ataatagcaa ttcctcgacg
     1081 actgcatagg gttacccccc tctccctccc ccccccctaa cgttactggc cgaagccgct
     1141 tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg
     1201 gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt
     1261 cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg
     1321 aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac
     1381 ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg
     1441 cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct
     1501 caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg
     1561 atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa aacgtctagg
     1621 ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataat ggccacaacc
     1681 atggtgagca agcagatcct gaagaacacc ggcctgcagg agatcatgag cttcaaggtg
     1741 aacctggagg gcgtggtgaa caaccacgtg ttcaccatgg agggctgcgg caagggcaac
     1801 atcctgttcg gcaaccagct ggtgcagatc cgcgtgacca agggcgcccc cctgcccttc
     1861 gccttcgaca tcctgagccc cgccttccag tacggcaacc gcaccttcac caagtacccc
     1921 gaggacatca gcgacttctt catccagagc ttccccgccg gcttcgtgta cgagcgcacc
     1981 ctgcgctacg aggacggcgg cctggtggag atccgcagcg acatcaacct gatcgaggag
     2041 atgttcgtgt accgcgtgga gtacaagggc cgcaacttcc ccaacgacgg ccccgtgatg
     2101 aagaagacca tcaccggcct gcagcccagc ttcgaggtgg tgtacatgaa cgacggcgtg
     2161 ctggtgggcc aggtgatcct ggtgtaccgc ctgaacagcg gcaagttcta cagctgccac
     2221 atgcgcaccc tgatgaagag caagggcgtg gtgaaggact tccccgagta ccacttcatc
     2281 cagcaccgcc tggagaagac ctacgtggag gacggcggct tcgtggagca gcacgagacc
     2341 gccatcgccc agctgaccag cctgggcaag cccctgggca gcctgcacga gtgggtgtaa
     2401 gaattccgta caaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg
     2461 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     2521 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     2581 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     2641 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     2701 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     2761 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     2821 tcatcgtcct ttccttggct gctcgcctat gttgccacct ggattctgcg cgggacgtcc
     2881 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     2941 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     3001 gccgcctccc cgcatcgata ccggctcgag acgggtggca tccctgtgac ccctccccag
     3061 tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc ctaataaaat
     3121 taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg tggagggggg
     3181 tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg gtctattggg
     3241 aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc tcctgggttc
     3301 aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc atgaccaggc
     3361 tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc aggctggtct
     3421 ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg ggattacagg
     3481 cgtgaaccac tgctcccttc cctgtccttc tcgagctgat tttgtaggta accacgtgcg
     3541 gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
     3601 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc
     3661 ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct
     3721 tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt
     3781 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     3841 agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     3901 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     3961 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga
     4021 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     4081 caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg
     4141 ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     4201 aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc
     4261 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     4321 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     4381 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     4441 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     4501 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     4561 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     4621 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     4681 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     4741 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     4801 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     4861 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     4921 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     4981 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     5041 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     5101 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     5161 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     5221 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     5281 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     5341 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     5401 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     5461 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     5521 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     5581 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     5641 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     5701 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     5761 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     5821 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     5881 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     5941 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     6001 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     6061 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     6121 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     6181 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     6241 aacgcggcct ttttacggtt cctggccttt t
//
