LOCUS       sequence-209549-        6302 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-mediated expression of last 50 amino acids of APLP1 protein 
            (APLP1ICD) and IRES-mediated co-expression of hrGFP to recognize 
            labelled cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6302)
  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 6302)
  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-APLP1ICD-IRES-hrGFP
FEATURES             Location/Qualifiers
     source          1..6302
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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             1622..1693
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     misc_feature    join(1729..2300,2301..2303,2304..2312)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1896..1913)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2123..2142
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2313..3032
                     /codon_start=1
                     /product="humanized Renilla green fluorescent protein"
                     /label=hrGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKQILKNTGLQEIMSFKVNLEGVVNNHVFTMEGCGKGNILFGN
                     QLVQIRVTKGAPLPFAFDILSPAFQYGNRTFTKYPEDISDFFIQSFPAGFVYERTLRYE
                     DGGLVEIRSDINLIEEMFVYRVEYKGRNFPNDGPVMKKTITGLQPSFEVVYMNDGVLVG
                     QVILVYRLNSGKFYSCHMRTLMKSKGVVKDFPEYHFIQHRLEKTYVEDGGFVEQHETAI
                     AQLTSLGKPLGSLHEWV"
     primer_bind     complement(3062..3081)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3063..3081)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3089..3565
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3288..3307)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3605..3745
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3605..3734
                     /label=AAV2 ITR
     rep_origin      3820..4275
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3907..3926)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4117..4138
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4292..4311)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4411..4433
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4471..4489)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4557..4661
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4662..4730,4731..5522)
                     /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(4880..4899)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5693..6281
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6182..6201
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttacggttcc tggccttttg ctggcctttt gctcacatgt cctgcaggca gctgcgcgct
       61 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggcgg 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 tccgaattcg ccaccatgct
     1441 gctcctgcgc aggaagaagc cctacggggc tatcagccat ggcgtggtgg aggtggaccc
     1501 catgctgacc ctggaggagc agcagctccg cgaactgcag cggcacggct atgagaaccc
     1561 cacttaccgc ttcctggagg aacgaccctg actcttcctg gaggaacgac cctgactcga
     1621 ggactacaag gatgacgatg acaaggatta caaagacgac gatgataagg actataagga
     1681 tgatgacgac aaataatagc aattcctcga cgactgcata gggttacccc cctctccctc
     1741 ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta
     1801 tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc
     1861 tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct
     1921 gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt
     1981 agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa
     2041 gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg
     2101 gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga
     2161 tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac
     2221 atgtgtttag tcgaggttaa aaaacgtcta ggccccccga accacgggga cgtggttttc
     2281 ctttgaaaaa cacgatgata atggccacaa ccatggtgag caagcagatc ctgaagaaca
     2341 ccggcctgca ggagatcatg agcttcaagg tgaacctgga gggcgtggtg aacaaccacg
     2401 tgttcaccat ggagggctgc ggcaagggca acatcctgtt cggcaaccag ctggtgcaga
     2461 tccgcgtgac caagggcgcc cccctgccct tcgccttcga catcctgagc cccgccttcc
     2521 agtacggcaa ccgcaccttc accaagtacc ccgaggacat cagcgacttc ttcatccaga
     2581 gcttccccgc cggcttcgtg tacgagcgca ccctgcgcta cgaggacggc ggcctggtgg
     2641 agatccgcag cgacatcaac ctgatcgagg agatgttcgt gtaccgcgtg gagtacaagg
     2701 gccgcaactt ccccaacgac ggccccgtga tgaagaagac catcaccggc ctgcagccca
     2761 gcttcgaggt ggtgtacatg aacgacggcg tgctggtggg ccaggtgatc ctggtgtacc
     2821 gcctgaacag cggcaagttc tacagctgcc acatgcgcac cctgatgaag agcaagggcg
     2881 tggtgaagga cttccccgag taccacttca tccagcaccg cctggagaag acctacgtgg
     2941 aggacggcgg cttcgtggag cagcacgaga ccgccatcgc ccagctgacc agcctgggca
     3001 agcccctggg cagcctgcac gagtgggtgt aatagggtac caggtaagtg tacccaattc
     3061 gccctatagt gagtcgtatt agatctacgg gtggcatccc tgtgacccct ccccagtgcc
     3121 tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag
     3181 ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt
     3241 atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc
     3301 aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc
     3361 gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga ccaggctcag
     3421 ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc tggtctccaa
     3481 ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat tacaggcgtg
     3541 aaccactgct cccttccctg tccttctgat tttgtaggta accacgtgcg gaccgagcgg
     3601 ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     3661 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
     3721 gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg
     3781 tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat
     3841 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag
     3901 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     3961 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4021 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4081 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4141 caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
     4201 tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     4261 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     4321 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     4381 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     4441 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     4501 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     4561 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     4621 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     4681 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     4741 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     4801 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     4861 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     4921 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     4981 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     5041 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     5101 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     5161 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     5221 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     5281 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     5341 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     5401 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     5461 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     5521 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     5581 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     5641 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     5701 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     5761 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     5821 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     5881 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5941 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6001 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6061 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     6121 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6181 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     6241 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     6301 tt
//
