LOCUS       sequence-345689-        5659 bp    DNA     circular SYN 15-APR-2025
DEFINITION  pAAV plasmid expressing an mCherry.3xFLAG.NOS1AP396-503 fusion 
            protein under the hSyn promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5659)
  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 5659)
  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: 
            "EBioMedicine."; date: "2021-08-26"; volume: "71:103565. doi"; 
            pages: " 10.1016/j.ebiom.2021.103565"
COMMENT     Sequence Label: pAAV-hSyn-mCherry.3xFLAG.NOS1AP396-503-WPRE
FEATURES             Location/Qualifiers
     source          1..5659
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        104..551
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      583..592
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             589..1296
                     /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(734..752)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1002..1021
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1303..1368
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    1715..2303
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1768..1788)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2186..2197)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2335..2811
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2534..2553)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2851..2991
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2851..2980
                     /label=AAV2 ITR
     rep_origin      3066..3521
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3153..3172)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3363..3384
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3538..3557)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3657..3679
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3717..3735)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3803..3907
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3908..3976,3977..4768)
                     /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(4126..4145)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4939..5527
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5428..5447
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(5589..5659,1..59)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta ggggttcctg
       61 cggccgcacg cgtgtgtcta gactgcagag ggccctgcgt atgagtgcaa gtgggtttta
      121 ggaccaggat gaggcggggt gggggtgcct acctgacgac cgaccccgac ccactggaca
      181 agcacccaac ccccattccc caaattgcgc atcccctatc agagaggggg aggggaaaca
      241 ggatgcggcg aggcgcgtgc gcactgccag cttcagcacc gcggacagtg ccttcgcccc
      301 cgcctggcgg cgcgcgccac cgccgcctca gcactgaagg cgcgctgacg tcactcgccg
      361 gtcccccgca aactcccctt cccggccacc ttggtcgcgt ccgcgccgcc gccggcccag
      421 ccggaccgca ccacgcgagg cgcgagatag gggggcacgg gcgcgaccat ctgcgctgcg
      481 gcgccggcga ctcagcgctg cctcagtctg cggtgggcag cggaggagtc gtgtcgtgcc
      541 tgagagcgca gtcgagaagg taccggatcc tctagagtcg acgccaccat ggtgagcaag
      601 ggcgaggagg ataacatggc catcatcaag gagttcatgc gcttcaaggt gcacatggag
      661 ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag
      721 ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac
      781 atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc cgccgacatc
      841 cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc
      901 gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc
      961 tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag
     1021 accatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc cctgaagggc
     1081 gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgctga ggtcaagacc
     1141 acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa catcaagttg
     1201 gacatcacct cccacaacga ggactacacc atcgtggaac agtacgaacg cgccgagggc
     1261 cgccactcca ccggcggcat ggacgagctg tacaaggaat tcgactacaa agaccatgac
     1321 ggtgattata aagatcatga tattgattac aaggatgacg atgacaagct agctcactca
     1381 ccactgctgg gcgctggctt ggctgacttt gcccacccag cgggcagccc tttaggtagg
     1441 catgactgct tggtgaagct ggagtgcttt cgcttcctcc cacctgagga tacccagccg
     1501 atgatggcac agggtgagcc gctcctaggt ggcctggagc tcatcaagtt ccgagagtcg
     1561 ggcatcgcct ctgagtatga gtccaacaca gacgaaagcg aggagcgtga ctcatggtca
     1621 caggaagagc tgccacgcct gctcaatgtc ttacagcggc aggagttggg tgacagtttg
     1681 gatgatgaga tcgccgtgta gtaagcttat cgataatcaa cctctggatt acaaaatttg
     1741 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     1801 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     1861 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     1921 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     1981 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     2041 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     2101 gtcggggaaa tcatcgtcct ttccttggct gctcgcctat gttgccacct ggattctgcg
     2161 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     2221 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     2281 ctccctttgg gccgcctccc cgcatcgata ccgagcgctg ctcgagagat ctacgggtgg
     2341 catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc
     2401 accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat
     2461 aatattatgg ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt
     2521 agggcctgcg gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact
     2581 gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc agcctcccga gttgttggga
     2641 ttccaggcat gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc
     2701 accatattgg ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct
     2761 cccaaattgc tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgattttg
     2821 taggtaacca cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc
     2881 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
     2941 gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga
     3001 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc
     3061 atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3121 gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct
     3181 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     3241 atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag
     3301 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     3361 tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga
     3421 tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa
     3481 atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac
     3541 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     3601 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     3661 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     3721 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     3781 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     3841 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     3901 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     3961 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     4021 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     4081 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     4141 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     4201 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     4261 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     4321 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     4381 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     4441 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     4501 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     4561 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     4621 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     4681 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     4741 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     4801 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     4861 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     4921 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     4981 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     5041 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     5101 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     5161 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     5221 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     5281 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     5341 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     5401 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     5461 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     5521 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     5581 tcacatgtcc tgcaggcagc tgcgcgctcg ctcgctcact gaggccgccc gggcgtcggg
     5641 cgacctttgg tcgcccggc
//
