LOCUS       sequence-209528-        6297 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-mediated expression of AICD (last 50 amino acids of APP) with 
            Nuclear Localisation Signal (NLS: CCAAAAAAGAAGAGAAAGGTA) at C-term 
            end and IRES-mediated co-expression of hrGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6297)
  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 6297)
  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-AICD-NLS-IRES-hrGFP
FEATURES             Location/Qualifiers
     source          1..6297
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   41..170
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        246..549
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        550..753
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     704..724
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     750..774
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          902..1280
                     /label=chimeric intron
                     /note="fusion between human cytomegalovirus intron A and 
                     human beta-globin intron 2"
     promoter        1351..1369
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             1587..1607
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1617..1688
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     misc_feature    join(1724..2295,2296..2298,2299..2307)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1891..1908)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2118..2137
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2308..3027
                     /codon_start=1
                     /product="humanized Renilla green fluorescent protein"
                     /label=hrGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKQILKNTGLQEIMSFKVNLEGVVNNHVFTMEGCGKGNILFGN
                     QLVQIRVTKGAPLPFAFDILSPAFQYGNRTFTKYPEDISDFFIQSFPAGFVYERTLRYE
                     DGGLVEIRSDINLIEEMFVYRVEYKGRNFPNDGPVMKKTITGLQPSFEVVYMNDGVLVG
                     QVILVYRLNSGKFYSCHMRTLMKSKGVVKDFPEYHFIQHRLEKTYVEDGGFVEQHETAI
                     AQLTSLGKPLGSLHEWV"
     primer_bind     complement(3057..3076)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3058..3076)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3084..3560
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3283..3302)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3600..3740
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3600..3729
                     /label=AAV2 ITR
     rep_origin      3815..4270
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3902..3921)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4112..4133
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4287..4306)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4406..4428
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4466..4484)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4552..4656
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4657..4725,4726..5517)
                     /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(4875..4894)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5688..6276
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6177..6196
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttacggttcc tggccttttg ctggcctttt gctcacatgt cctgcaggca gctgcgcgct
       61 cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag
      121 cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct gcggccgcac
      181 gcgtctagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      241 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      301 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      361 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      421 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      481 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      541 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      601 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      661 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      721 cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg
      781 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgc
      841 ggattcgaat cccggccggg aacggtgcat tggaacgcgg attccccgtg ccaagagtga
      901 cgtaagtacc gcctatagag tctataggcc cacaaaaaat gctttcttct tttaatatac
      961 ttttttgttt atcttatttc taatactttc cctaatctct ttctttcagg gcaataatga
     1021 tacaatgtat catgcctctt tgcaccattc taaagaataa cagtgataat ttctgggtta
     1081 aggcaatagc aatatttctg catataaata tttctgcata taaattgtaa ctgatgtaag
     1141 aggtttcata ttgctaatag cagctacaat ccagctacca ttctgctttt attttatggt
     1201 tgggataagg ctggattatt ctgagtccaa gctaggccct tttgctaatc atgttcatac
     1261 ctcttatctt cctcccacag ctcctgggca acgtgctggt ctgtgtgctg gcccatcact
     1321 ttggcaaaga attgggattc gaacatcgat aattaaccct cactaaaggg aacaaaagct
     1381 ggagctccac cgcggtggcg gccgctctag cccgggcgga tccgaattcc accatggtga
     1441 tgctgaagaa gaaacagtac acatccattc atcatggtgt ggtggaggtt gacgccgctg
     1501 tcaccccaga ggagcgccac ctgtccaaga tgcagcagaa cggctacgaa aatccaacct
     1561 acaagttctt tgagcagatg cagaacccaa aaaagaagag aaaggtatga ctcgaggact
     1621 acaaggatga cgatgacaag gattacaaag acgacgatga taaggactat aaggatgatg
     1681 acgacaaata atagcaattc ctcgacgact gcatagggtt acccccctct ccctcccccc
     1741 cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt
     1801 tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct
     1861 tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga
     1921 atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga
     1981 ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac
     2041 gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag
     2101 ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc
     2161 agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg
     2221 tttagtcgag gttaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg
     2281 aaaaacacga tgataatggc cacaaccatg gtgagcaagc agatcctgaa gaacaccggc
     2341 ctgcaggaga tcatgagctt caaggtgaac ctggagggcg tggtgaacaa ccacgtgttc
     2401 accatggagg gctgcggcaa gggcaacatc ctgttcggca accagctggt gcagatccgc
     2461 gtgaccaagg gcgcccccct gcccttcgcc ttcgacatcc tgagccccgc cttccagtac
     2521 ggcaaccgca ccttcaccaa gtaccccgag gacatcagcg acttcttcat ccagagcttc
     2581 cccgccggct tcgtgtacga gcgcaccctg cgctacgagg acggcggcct ggtggagatc
     2641 cgcagcgaca tcaacctgat cgaggagatg ttcgtgtacc gcgtggagta caagggccgc
     2701 aacttcccca acgacggccc cgtgatgaag aagaccatca ccggcctgca gcccagcttc
     2761 gaggtggtgt acatgaacga cggcgtgctg gtgggccagg tgatcctggt gtaccgcctg
     2821 aacagcggca agttctacag ctgccacatg cgcaccctga tgaagagcaa gggcgtggtg
     2881 aaggacttcc ccgagtacca cttcatccag caccgcctgg agaagaccta cgtggaggac
     2941 ggcggcttcg tggagcagca cgagaccgcc atcgcccagc tgaccagcct gggcaagccc
     3001 ctgggcagcc tgcacgagtg ggtgtaatag ggtaccaggt aagtgtaccc aattcgccct
     3061 atagtgagtc gtattagatc tacgggtggc atccctgtga cccctcccca gtgcctctcc
     3121 tggccctgga agttgccact ccagtgccca ccagccttgt cctaataaaa ttaagttgca
     3181 tcattttgtc tgactaggtg tccttctata atattatggg gtggaggggg gtggtatgga
     3241 gcaaggggca agttgggaag acaacctgta gggcctgcgg ggtctattgg gaaccaagct
     3301 ggagtgcagt ggcacaatct tggctcactg caatctccgc ctcctgggtt caagcgattc
     3361 tcctgcctca gcctcccgag ttgttgggat tccaggcatg catgaccagg ctcagctaat
     3421 ttttgttttt ttggtagaga cggggtttca ccatattggc caggctggtc tccaactcct
     3481 aatctcaggt gatctaccca ccttggcctc ccaaattgct gggattacag gcgtgaacca
     3541 ctgctccctt ccctgtcctt ctgattttgt aggtaaccac gtgcggaccg agcggccgca
     3601 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     3661 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg
     3721 agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     3781 tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc
     3841 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc
     3901 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     3961 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     4021 aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     4081 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     4141 ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat
     4201 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     4261 tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     4321 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     4381 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     4441 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     4501 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     4561 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     4621 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     4681 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     4741 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     4801 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     4861 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     4921 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     4981 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     5041 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     5101 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     5161 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     5221 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     5281 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     5341 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     5401 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     5461 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     5521 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     5581 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5641 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     5701 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     5761 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     5821 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     5881 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     5941 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     6001 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     6061 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     6121 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     6181 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     6241 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttt
//
