LOCUS       sequence-401485-        6442 bp    DNA     circular SYN 14-APR-2025
DEFINITION  AAV vector for miniDq expression under the control of human synapsin
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6442)
  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 6442)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-P2A-mCherry
FEATURES             Location/Qualifiers
     source          1..6442
                     /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             1794..1850
                     /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             1851..2558
                     /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(1993..2011)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2261..2280
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2569..3157
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2622..2642)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3040..3051)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3189..3665
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3388..3407)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3705..3845
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3705..3834
                     /label=AAV2 ITR
     rep_origin      3920..4375
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4007..4026)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4217..4238
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4392..4411)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4511..4533
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        4657..4761
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4762..4830,4831..5622)
                     /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(4980..4999)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5793..6381
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6282..6301
                     /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 ttggccacta
     1801 acttctccct gttgaaacaa gcaggggatg tcgaagagaa tcccgggcca gtgagcaagg
     1861 gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg
     1921 gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg
     1981 gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca
     2041 tcctgtcccc tcagttcatg tacggctcca aggcctacgt gaagcacccc gccgacatcc
     2101 ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg
     2161 aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct
     2221 acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga
     2281 ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg
     2341 agatcaagca gaggctgaag ctgaaggacg gcggccacta cgacgctgag gtcaagacca
     2401 cctacaaggc caagaagccc gtgcagctgc ccggcgccta caacgtcaac atcaagttgg
     2461 acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc gccgagggcc
     2521 gccactccac cggcggcatg gacgagctgt acaagtaagc ttatcgataa tcaacctctg
     2581 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     2641 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     2701 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     2761 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     2821 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     2881 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     2941 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc
     3001 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     3061 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     3121 cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgagc gctgctcgag
     3181 agatctacgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg
     3241 ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact
     3301 aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg
     3361 ggaagacaac ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac
     3421 aatcttggct cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc
     3481 ccgagttgtt gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt
     3541 agagacgggg tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct
     3601 acccaccttg gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg
     3661 tccttctgat tttgtaggta accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat
     3721 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt
     3781 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct
     3841 gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     3901 acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     3961 gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc
     4021 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     4081 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt
     4141 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     4201 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg
     4261 ggctattctt ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag
     4321 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg
     4381 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     4441 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     4501 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     4561 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     4621 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4681 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4741 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     4801 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     4861 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     4921 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     4981 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     5041 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     5101 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     5161 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     5221 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     5281 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     5341 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     5401 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     5461 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     5521 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     5581 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     5641 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     5701 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     5761 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     5821 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     5881 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     5941 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     6001 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     6061 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     6121 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     6181 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     6241 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     6301 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     6361 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     6421 tgctggcctt ttgctcacat gt
//
