LOCUS       sequence-345686-        5650 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pAAV plasmid expressing an mCherry.3xFLAG.NOS1AP396-500 fusion 
            protein under the hSyn promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5650)
  AUTHORS   Freudenberg F, Candemir E, Chen X, Li LL, Esen-Sehir D, Schenk N, 
            Kinoshita M, Grunewald L, Frerichs V, Fattakhov N, Manchen J, Bikas 
            S, Kumar A, OLeary A, Slattery DA, von Engelhardt J, Courtney MJ, 
            Reif A
  TITLE     Hippocampal overexpression of NOS1AP promotes endophenotypes related
            to mental disorders.
  JOURNAL   EBioMedicine. 2021 Aug 26;71:103565. doi: 
            10.1016/j.ebiom.2021.103565.
  PUBMED    34455393
REFERENCE   2  (bases 1 to 5650)
  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: 
            "EBioMedicine."; date: "2021-08-26"; volume: "71:103565. doi"; 
            pages: " 10.1016/j.ebiom.2021.103565"
COMMENT     Sequence Label: pAAV-hSyn-mCherry.3xFLAG.NOS1AP396-500-WPRE
FEATURES             Location/Qualifiers
     source          1..5650
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   651..780
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        825..1272
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      1304..1313
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1310..2017
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1455..1473)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1723..1742
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2024..2089
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    2427..3015
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2480..2500)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2898..2909)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3047..3523
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3246..3265)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3563..3703
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3563..3692
                     /label=AAV2 ITR
     rep_origin      3778..4233
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3865..3884)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4075..4096
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4250..4269)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4369..4391
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4429..4447)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4515..4619
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4620..4688,4689..5480)
                     /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(4838..4857)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt cctgcaggca
      661 gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt ggtcgcccgg
      721 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
      781 gcggccgcac gcgtgtgtct agactgcaga gggccctgcg tatgagtgca agtgggtttt
      841 aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga cccactggac
      901 aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg gaggggaaac
      961 aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt gccttcgccc
     1021 ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac gtcactcgcc
     1081 ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc cgccggccca
     1141 gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca tctgcgctgc
     1201 ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt cgtgtcgtgc
     1261 ctgagagcgc agtcgagaag gtaccggatc ctctagagtc gacgccacca tggtgagcaa
     1321 gggcgaggag gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga
     1381 gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga
     1441 gggcacccag accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga
     1501 catcctgtcc cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat
     1561 ccccgactac ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt
     1621 cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat
     1681 ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa
     1741 gaccatgggc tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg
     1801 cgagatcaag cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac
     1861 cacctacaag gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt
     1921 ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg
     1981 ccgccactcc accggcggca tggacgagct gtacaaggaa ttcgactaca aagaccatga
     2041 cggtgattat aaagatcatg atattgatta caaggatgac gatgacaagc tagctcactc
     2101 accactgctg ggcgctggct tggctgactt tgcccaccca gcgggcagcc ctttaggtag
     2161 gcatgactgc ttggtgaagc tggagtgctt tcgcttcctc ccacctgagg atacccagcc
     2221 gatgatggca cagggtgagc cgctcctagg tggcctggag ctcatcaagt tccgagagtc
     2281 gggcatcgcc tctgagtatg agtccaacac agacgaaagc gaggagcgtg actcatggtc
     2341 acaggaagag ctgccacgcc tgctcaatgt cttacagcgg caggagttgg gtgacagttt
     2401 ggatgatgag tagtaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat
     2461 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     2521 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     2581 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     2641 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     2701 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     2761 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     2821 aatcatcgtc ctttccttgg ctgctcgcct atgttgccac ctggattctg cgcgggacgt
     2881 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     2941 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     3001 gggccgcctc cccgcatcga taccgagcgc tgctcgagag atctacgggt ggcatccctg
     3061 tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct
     3121 tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat
     3181 ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg
     3241 cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc
     3301 cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg gattccaggc
     3361 atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt
     3421 ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt
     3481 gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt tgtaggtaac
     3541 cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg
     3601 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     3661 cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat
     3721 tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg
     3781 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     3841 cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     3901 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     3961 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
     4021 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     4081 tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt gatttataag
     4141 ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     4201 cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct
     4261 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     4321 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     4381 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     4441 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     4501 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4561 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     4621 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     4681 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     4741 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     4801 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     4861 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     4921 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     4981 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     5041 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     5101 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     5161 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     5221 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     5281 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     5341 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     5401 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     5461 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     5521 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     5581 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     5641 aaggatcttc
//
