LOCUS       sequence-209555-        6299 bp    DNA     circular SYN 15-APR-2025
DEFINITION  AAV-mediated expression of AICD (last 50 amino acids of APP) with 
            Nuclear Export Signal (NES: CTCCCTCCACTAGAGCGACTAACCTTA) 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 6299)
  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 6299)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-NES-IRES-hrGFP
FEATURES             Location/Qualifiers
     source          1..6299
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        264..567
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        568..771
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     722..742
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     768..792
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          920..1298
                     /label=chimeric intron
                     /note="fusion between human cytomegalovirus intron A and 
                     human beta-globin intron 2"
     promoter        1369..1387
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      1442..1451
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1601..1627
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             1637..1708
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     misc_feature    join(1744..2315,2316..2318,2319..2327)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1911..1928)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2138..2157
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2328..3047
                     /codon_start=1
                     /product="humanized Renilla green fluorescent protein"
                     /label=hrGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKQILKNTGLQEIMSFKVNLEGVVNNHVFTMEGCGKGNILFGN
                     QLVQIRVTKGAPLPFAFDILSPAFQYGNRTFTKYPEDISDFFIQSFPAGFVYERTLRYE
                     DGGLVEIRSDINLIEEMFVYRVEYKGRNFPNDGPVMKKTITGLQPSFEVVYMNDGVLVG
                     QVILVYRLNSGKFYSCHMRTLMKSKGVVKDFPEYHFIQHRLEKTYVEDGGFVEQHETAI
                     AQLTSLGKPLGSLHEWV"
     primer_bind     complement(3077..3096)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3078..3096)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3104..3580
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3303..3322)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3620..3760
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3620..3749
                     /label=AAV2 ITR
     rep_origin      3835..4290
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3922..3941)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4132..4153
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4307..4326)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4426..4448
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4486..4504)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4572..4676
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4677..4745,4746..5537)
                     /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(4895..4914)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5708..6296
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6197..6216
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtcc
       61 tgcaggcagc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc
      121 ccgggcggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag
      181 gggttcctgc ggccgcacgc gtctagttat taatagtaat caattacggg gtcattagtt
      241 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      301 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      361 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
      421 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      481 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      541 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
      601 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
      661 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
      721 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg
      781 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg
      841 gaccgatcca gcctccgcgg attcgaatcc cggccgggaa cggtgcattg gaacgcggat
      901 tccccgtgcc aagagtgacg taagtaccgc ctatagagtc tataggccca caaaaaatgc
      961 tttcttcttt taatatactt ttttgtttat cttatttcta atactttccc taatctcttt
     1021 ctttcagggc aataatgata caatgtatca tgcctctttg caccattcta aagaataaca
     1081 gtgataattt ctgggttaag gcaatagcaa tatttctgca tataaatatt tctgcatata
     1141 aattgtaact gatgtaagag gtttcatatt gctaatagca gctacaatcc agctaccatt
     1201 ctgcttttat tttatggttg ggataaggct ggattattct gagtccaagc taggcccttt
     1261 tgctaatcat gttcatacct cttatcttcc tcccacagct cctgggcaac gtgctggtct
     1321 gtgtgctggc ccatcacttt ggcaaagaat tgggattcga acatcgataa ttaaccctca
     1381 ctaaagggaa caaaagctgg agctccaccg cggtggcggc cgctctagcc cgggcggatc
     1441 cgccaccatg gtgatgctga agaagaaaca gtacacatcc attcatcatg gtgtggtgga
     1501 ggttgacgcc gctgtcaccc cagaggagcg ccacctgtcc aagatgcagc agaacggcta
     1561 cgaaaatcca acctacaagt tctttgagca gatgcagaac ctccctccac tagagcgact
     1621 aaccttatga ctcgaggact acaaggatga cgatgacaag gattacaaag acgacgatga
     1681 taaggactat aaggatgatg acgacaaata atagcaattc ctcgacgact gcatagggtt
     1741 acccccctct ccctcccccc cccctaacgt tactggccga agccgcttgg aataaggccg
     1801 gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc
     1861 ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa
     1921 aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag
     1981 acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg
     2041 cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg
     2101 ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa
     2161 caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg
     2221 gtgcacatgc tttacatgtg tttagtcgag gttaaaaaac gtctaggccc cccgaaccac
     2281 ggggacgtgg ttttcctttg aaaaacacga tgataatggc cacaaccatg gtgagcaagc
     2341 agatcctgaa gaacaccggc ctgcaggaga tcatgagctt caaggtgaac ctggagggcg
     2401 tggtgaacaa ccacgtgttc accatggagg gctgcggcaa gggcaacatc ctgttcggca
     2461 accagctggt gcagatccgc gtgaccaagg gcgcccccct gcccttcgcc ttcgacatcc
     2521 tgagccccgc cttccagtac ggcaaccgca ccttcaccaa gtaccccgag gacatcagcg
     2581 acttcttcat ccagagcttc cccgccggct tcgtgtacga gcgcaccctg cgctacgagg
     2641 acggcggcct ggtggagatc cgcagcgaca tcaacctgat cgaggagatg ttcgtgtacc
     2701 gcgtggagta caagggccgc aacttcccca acgacggccc cgtgatgaag aagaccatca
     2761 ccggcctgca gcccagcttc gaggtggtgt acatgaacga cggcgtgctg gtgggccagg
     2821 tgatcctggt gtaccgcctg aacagcggca agttctacag ctgccacatg cgcaccctga
     2881 tgaagagcaa gggcgtggtg aaggacttcc ccgagtacca cttcatccag caccgcctgg
     2941 agaagaccta cgtggaggac ggcggcttcg tggagcagca cgagaccgcc atcgcccagc
     3001 tgaccagcct gggcaagccc ctgggcagcc tgcacgagtg ggtgtaatag ggtaccaggt
     3061 aagtgtaccc aattcgccct atagtgagtc gtattagatc tacgggtggc atccctgtga
     3121 cccctcccca gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt
     3181 cctaataaaa ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg
     3241 gtggaggggg gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg
     3301 ggtctattgg gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc
     3361 ctcctgggtt caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg
     3421 catgaccagg ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc
     3481 caggctggtc tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct
     3541 gggattacag gcgtgaacca ctgctccctt ccctgtcctt ctgattttgt aggtaaccac
     3601 gtgcggaccg agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc
     3661 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg
     3721 gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt
     3781 ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc
     3841 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3901 ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3961 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     4021 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     4081 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     4141 tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga
     4201 ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     4261 attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg
     4321 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     4381 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     4441 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     4501 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     4561 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     4621 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     4681 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     4741 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     4801 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     4861 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     4921 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     4981 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     5041 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     5101 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     5161 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     5221 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     5281 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     5341 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     5401 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     5461 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     5521 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     5581 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     5641 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     5701 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     5761 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     5821 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     5881 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     5941 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     6001 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     6061 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     6121 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     6181 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     6241 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaac
//
