LOCUS       sequence-401494-        6274 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector for miniDi expression under the control of human synapsin
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6274)
  AUTHORS   Miyake T, Tanaka K, Inoue Y, Nagai Y, Nishimura R, Seta T, Nakagawa 
            S, Inoue KI, Hasegawa E, Minamimoto T, Doi M
  TITLE     Size-reduced DREADD derivatives for AAV-assisted multimodal 
            chemogenetic control of neuronal activity and behavior.
  JOURNAL   Cell Rep Methods. 2024 Oct 21;4(10):100881. doi: 
            10.1016/j.crmeth.2024.100881.
  PUBMED    39437713
REFERENCE   2  (bases 1 to 6274)
  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: "Cell Rep 
            Methods."; date: "2024-10-21"; volume: "4(10):100881. doi"; pages: "
            10.1016/j.crmeth.2024.100881"
COMMENT     Sequence Label: pAAV-hSyn-miniDi-P2A-mCherry
FEATURES             Location/Qualifiers
     source          1..6274
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(57..125,126..917)
                     /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(275..294)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1088..1676
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1577..1596
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   1738..1867
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        1912..2359
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      2373..2382
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3363..3419
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             3420..4127
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(3562..3580)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3830..3849
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    4138..4726
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4191..4211)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4609..4620)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4758..5234
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4957..4976)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5274..5414
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5274..5403
                     /label=AAV2 ITR
     rep_origin      5489..5944
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5576..5595)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5786..5807
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5961..5980)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6080..6102
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6140..6158)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(6226..6274,1..56)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
       61 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
      121 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
      181 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
      241 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
      301 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
      361 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
      421 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
      481 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
      541 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
      601 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
      661 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
      721 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
      781 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
      841 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
      901 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
      961 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     1021 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     1081 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     1141 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     1201 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     1261 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     1321 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     1381 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     1441 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     1501 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     1561 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     1621 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     1681 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtcct
     1741 gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg ggcgtcgggc gacctttggt
     1801 cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg
     1861 ggttcctgcg gccgcacgcg tgtgtctaga ctgcagaggg ccctgcgtat gagtgcaagt
     1921 gggttttagg accaggatga ggcggggtgg gggtgcctac ctgacgaccg accccgaccc
     1981 actggacaag cacccaaccc ccattcccca aattgcgcat cccctatcag agagggggag
     2041 gggaaacagg atgcggcgag gcgcgtgcgc actgccagct tcagcaccgc ggacagtgcc
     2101 ttcgcccccg cctggcggcg cgcgccaccg ccgcctcagc actgaaggcg cgctgacgtc
     2161 actcgccggt cccccgcaaa ctccccttcc cggccacctt ggtcgcgtcc gcgccgccgc
     2221 cggcccagcc ggaccgcacc acgcgaggcg cgagataggg gggcacgggc gcgaccatct
     2281 gcgctgcggc gccggcgact cagcgctgcc tcagtctgcg gtgggcagcg gaggagtcgt
     2341 gtcgtgcctg agagcgcagt cgagaaggta ccgccaccat ggccaacttc acacctgtca
     2401 atggcagctc gggcaatcag tccgtgcgcc tggtcacgtc atcatcccac aatcgctatg
     2461 agacggtgga aatggtcttc attgccacag tgacaggctc cctgagcctg gtgactgtcg
     2521 tgggcaacat cctggtgatg ctgtccatca aggtcaacag gcagctgcag acagtcaaca
     2581 actacttcct cttcagcctg gcgtgtgctg atctcatcat aggcgccttc tccatgaacc
     2641 tctacaccgt gtacatcatc aagggctact ggcccctggg cgccgtggtc tgcgacctgt
     2701 ggctggccct ggactgcgtg gtgagcaacg cctccgtcat gaaccttctc atcatcagct
     2761 ttgaccgcta cttctgcgtc accaagcctc tcacctaccc tgcccggcgc accaccaaga
     2821 tggcaggcct catgattgct gctgcctggg tactgtcctt cgtgctctgg gcgcctgcca
     2881 tcttgttctg gcagtttgtg gtgggtaagc ggacggtgcc cgacaaccag tgcttcatcc
     2941 agttcctgtc caacccagca gtgacctttg gcacagccat tgctggcttc tacctgcctg
     3001 tggtcatcat gacggtgctg tacatccaca tctccctggc cagtcgcagc cgagtccaca
     3061 agcaccggcc cgagcagaac accatctctc ggcagatggc ggcccgggag cgcaaagtga
     3121 cacgaacgat ctttgccatt ctgctggcct tcatcctcac ctggacgccc tacaacgtca
     3181 tggtcctggt gaacaccttc tgccagagct gcatccctga cacggtgtgg tccattggct
     3241 actggctctg ctacgtcaac agcaccatca accctgcctg ctatgctctg tgcaacgcca
     3301 cctttaaaaa gaccttccgg cacctgctgc tgtgccagta tcggaacatc ggcactgcca
     3361 gggccactaa cttctccctg ttgaaacaag caggggatgt cgaagagaat cccgggccag
     3421 tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc
     3481 acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
     3541 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg
     3601 cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg
     3661 ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga
     3721 tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg
     3781 agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc
     3841 agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc
     3901 tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg
     3961 tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca
     4021 tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg
     4081 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtaagct tatcgataat
     4141 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     4201 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     4261 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     4321 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     4381 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     4441 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     4501 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc
     4561 tatgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     4621 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     4681 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg ataccgagcg
     4741 ctgctcgaga gatctacggg tggcatccct gtgacccctc cccagtgcct ctcctggccc
     4801 tggaagttgc cactccagtg cccaccagcc ttgtcctaat aaaattaagt tgcatcattt
     4861 tgtctgacta ggtgtccttc tataatatta tggggtggag gggggtggta tggagcaagg
     4921 ggcaagttgg gaagacaacc tgtagggcct gcggggtcta ttgggaacca agctggagtg
     4981 cagtggcaca atcttggctc actgcaatct ccgcctcctg ggttcaagcg attctcctgc
     5041 ctcagcctcc cgagttgttg ggattccagg catgcatgac caggctcagc taatttttgt
     5101 ttttttggta gagacggggt ttcaccatat tggccaggct ggtctccaac tcctaatctc
     5161 aggtgatcta cccaccttgg cctcccaaat tgctgggatt acaggcgtga accactgctc
     5221 ccttccctgt ccttctgatt ttgtaggtaa ccacgtgcgg accgagcggc cgcaggaacc
     5281 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
     5341 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg
     5401 cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     5461 acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg
     5521 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct
     5581 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     5641 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     5701 gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     5761 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     5821 tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta
     5881 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
     5941 attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     6001 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     6061 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     6121 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     6181 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     6241 tgtttatttt tctaaataca ttcaaatatg tatc
//
