LOCUS       sequence-401492-        7296 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector for coexpression of miniDq and KORD under the control of 
            human synapsin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7296)
  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 7296)
  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-miniDq-mCherry-P2A-HA-KORD
FEATURES             Location/Qualifiers
     source          1..7296
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        175..622
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             1803..2507
                     /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(1945..1963)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2213..2232
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2508..2564
                     /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             2568..2594
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(3228..3239)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     misc_feature    3749..4337
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3802..3822)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4220..4231)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4380..4501
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4391..4410)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4445..4464
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4559..4699
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4559..4688
                     /label=AAV2 ITR
     rep_origin      4774..5229
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4861..4880)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5071..5092
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5246..5265)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5365..5387
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        5511..5615
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5616..5684,5685..6476)
                     /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(5834..5853)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6647..7235
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7136..7155
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtgtgtct agactgcaga gggccctgcg tatgagtgca
      181 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga
      241 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg
      301 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt
      361 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac
      421 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc
      481 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca
      541 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt
      601 cgtgtcgtgc ctgagagcgc agtcgagaag gtaccgccac catgaccttg cacaataaca
      661 gtacaacctc gcctttgttt ccaaacatca gctcctcctg gatacacagc ccctccgatg
      721 cagggctgcc cccgggaacc gtcactcatt tcggcagcta caatgtttct cgagcagctg
      781 gcaatttctc ctctccagac ggtaccaccg atgaccctct gggaggtcat accgtctggc
      841 aagtggtctt catcgctttc ttaacgggca tcctggcctt ggtgaccatc atcggcaaca
      901 tcctggtaat tgtgtcattt aaggtcaaca agcagctgaa gacggtcaac aactacttcc
      961 tcttaagcct ggcctgtgcc gatctgatta tcggggtcat ttcaatgaat ctgtttacga
     1021 cctacatcat catgaatcga tgggccttag ggaacttggc ctgtgacctc tggcttgcca
     1081 ttgactgcgt agccagcaat gcctctgtta tgaatcttct ggtcatcagc tttgacagat
     1141 acttttccat cacgaggccg ctcacgtacc gagccaaacg aacaacaaag agagccggtg
     1201 tgatgatcgg tctggcttgg gtcatctcct ttgtcctttg ggctcctgcc atcttgttct
     1261 ggcaatactt tgttggaaag agaactgtgc ctccgggaga gtgcttcatt cagttcctca
     1321 gtgagcccac cattactttt ggcacagcca tcgctggttt ttatatgcct gtcaccatta
     1381 tgactatttt atactggagg atctataagg aaactgaaaa gcgtaccaaa gagcttgctg
     1441 gcctgcagaa caccatctct aggatgtccc tggtcaagga gaagaaagcg gcccagaccc
     1501 tcagtgcgat cttgcttgcc ttcatcatca cttggacccc atacaacatc atggttctgg
     1561 tgaacacctt ttgtgacagc tgcataccca aaaccttttg gaatctgggc tactggctgt
     1621 gctacatcaa cagcaccgtg aaccccgtgt gctatgctct gtgcaacaaa acattcagaa
     1681 ccactttcaa gatgctgctg ctgtgccagt gtgacaaaaa aaagaggcgc aagcagcagt
     1741 accagcagag acagtcggtc atttttcaca agcgcgcacc cgagcaggcc ttggcggcta
     1801 gcgtgagcaa gggcgaggag gataacatgg ccatcatcaa ggagttcatg cgcttcaagg
     1861 tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc
     1921 gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc cccctgccct
     1981 tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac gtgaagcacc
     2041 ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag tgggagcgcg
     2101 tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg
     2161 gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtaa
     2221 tgcagaagaa gaccatgggc tgggaggcct cctccgagcg gatgtacccc gaggacggcg
     2281 ccctgaaggg cgagatcaag cagaggctga agctgaagga cggcggccac tacgacgctg
     2341 aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca
     2401 acatcaagtt ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgaac
     2461 gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaaggcc actaacttct
     2521 ccctgttgaa acaagcaggg gatgtcgaag agaatcccgg gccaatgtac ccatacgatg
     2581 ttccagatta cgctatggaa tccccgattc agatcttccg cggggagccg ggccctacct
     2641 gcgccccgag cgcctgcctg ccccccaaca gcagcgcctg gtttcccggc tgggccgagc
     2701 ccgacagcaa cggcagcgcc ggctcggagg acgcgcagct ggagcccgcg cacatctccc
     2761 cggccatccc ggtcatcatc acggcggtct actccgtagt gttcgtcgtg ggcttggtgg
     2821 gcaactcgct ggtcatgttc gtgatcatcc gatacacaaa gatgaagaca gcaaccaaca
     2881 tttacatatt taacctggct ttggcagatg ctttagttac tacaaccatg ccctttcaga
     2941 gtacggtcta cttgatgaac tcctggcctt ttggggatgt gctgtgcaag atagtaattt
     3001 ccattaatta ctacaacatg ttcaccagca tcttcacctt gaccatgatg agcgtggacc
     3061 gctacattgc cgtgtgccac cccgtgaagg ctttggactt ccgcacaccc ttgaaggcaa
     3121 agatcatcaa tatctgcatc tggctgctgt cgtcatctgt tggcatctct gcaatagtcc
     3181 ttggaggcac caaagtcagg gaagacgtcg atgtcattga gtgctccttg cagttcccag
     3241 atgatgacta ctcctggtgg gacctcttca tgaagatctg cgtcttcatc tttgccttcg
     3301 tgatccctgt cctcatcatc atcgtctgct acaccctgat gatcctgcgt ctcaagagcg
     3361 tccggctcct ttctggctcc cgagagaaag atcgcaacct gcgtaggatc accagactgg
     3421 tcctggtggt ggtggcagtc ttcgtcgtct gctggactcc cattcacata ttcatcctgg
     3481 tggaggctct ggggagcacc tcccacagca cagctgctct ctccagctat tacttctgca
     3541 tcgccttagg ctataccaac agtagcctga atcccattct ctacgccttt cttgatgaaa
     3601 acttcaagcg gtgtttccgg gacttctgct ttccactgaa gatgaggatg gagcggcaga
     3661 gcactagcag agtccgaaat acagttcagg accctgctta cctgagggac atcgatggga
     3721 tgaataaacc agtataaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag
     3781 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     3841 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     3901 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     3961 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     4021 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     4081 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     4141 gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     4201 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     4261 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     4321 ttgggccgcc tccccgcatc gataccgtcg acccgggcgg ccgcttcgag cagacatgat
     4381 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     4441 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     4501 taacaacaac aattgcattc attttatgtt tcaggttcag ggggagatgt ggggccgcag
     4561 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
     4621 gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga
     4681 gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     4741 atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg
     4801 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg
     4861 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     4921 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     4981 aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     5041 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     5101 tcaactctat ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt
     5161 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
     5221 ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     5281 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     5341 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     5401 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     5461 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     5521 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     5581 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     5641 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     5701 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     5761 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     5821 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     5881 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     5941 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     6001 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     6061 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     6121 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     6181 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     6241 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     6301 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     6361 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     6421 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     6481 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     6541 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     6601 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     6661 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     6721 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     6781 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     6841 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     6901 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     6961 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     7021 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     7081 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     7141 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     7201 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     7261 ggttcctggc cttttgctgg ccttttgctc acatgt
//
