LOCUS       sequence-355043-        7057 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector for Cre-dependent transgene expression of 
            Dreadd-hM3Dq-HA-P2A-tBFP in neurons under the control of the hSyn1 
            promoter (Kugler et al 2003).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7057)
  AUTHORS   Pouchelon G, Vergara J, McMahon J, Gorissen BL, Lin JD, 
            Vormstein-Schneider D, Niehaus JL, Burbridge TJ, Wester JC, Sherer 
            M, Fernandez-Otero M, Allaway KC, Pelkey K, Chittajallu R, McBain 
            CJ, Fan M, Nasse JS, Wildenberg GA, Fishell G, Dimidschstein J
  TITLE     A versatile viral toolkit for functional discovery in the nervous 
            system.
  JOURNAL   Cell Rep Methods. 2022 May 26;2(6):100225. doi: 
            10.1016/j.crmeth.2022.100225. eCollection 2022 Jun 20.
  PUBMED    35784651
REFERENCE   2  (bases 1 to 7057)
  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: "2022-05-26"; volume: "2(6):100225. doi"; pages: " 
            10.1016/j.crmeth.2022.100225. eCollection 2022 Jun 20"
COMMENT     Sequence Label: pAAV-VTKS2-Dreadd-hM3Dq-HA-P2A-tBFP
FEATURES             Location/Qualifiers
     source          1..7057
                     /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        158..605
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(629..662)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(713..746)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(767..1465)
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     CDS             complement(1466..1522)
                     /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             complement(3311..3337)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    3372..3405
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    3456..3489
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3498..4086
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3551..3571)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3969..3980)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4094..4318
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4320..4460
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4320..4449
                     /label=AAV2 ITR
     rep_origin      4535..4990
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4622..4641)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4832..4853
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5007..5026)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5126..5148
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        5272..5376
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5377..5445,5446..6237)
                     /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(5595..5614)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6408..6996
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6897..6916
                     /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 gctagactgc agagggccct gcgtatgagt gcaagtgggt tttaggacca
      181 ggatgaggcg gggtgggggt gcctacctga cgaccgaccc cgacccactg gacaagcacc
      241 caacccccat tccccaaatt gcgcatcccc tatcagagag ggggagggga aacaggatgc
      301 ggcgaggcgc gtgcgcactg ccagcttcag caccgcggac agtgccttcg cccccgcctg
      361 gcggcgcgcg ccaccgccgc ctcagcactg aaggcgcgct gacgtcactc gccggtcccc
      421 cgcaaactcc ccttcccggc caccttggtc gcgtccgcgc cgccgccggc ccagccggac
      481 cgcaccacgc gaggcgcgag ataggggggc acgggcgcga ccatctgcgc tgcggcgccg
      541 gcgactcagc gctgcctcag tctgcggtgg gcagcggagg agtcgtgtcg tgcctgagag
      601 cgcagtcaga tatcgtttaa actccggaat aacttcgtat aggatacttt atacgaagtt
      661 atgcagaatg gtagctggat tgtagctgct attagcaata tgaaacctct taataacttc
      721 gtatagcata cattatacga agttataacg cgccactagt gaattcttaa ttaagcttgt
      781 gccccagttt gctagggagg tcgcagtatc tggccactgc cacctcgtgc tgctcgacgt
      841 aggtctcgtt gttggcctcc ttgattcttt ccagtctgta gtccacatag tagacgccag
      901 gcatcttgag gttcttagcg ggtttcttgg atctatatgt ggtcttgatg tttgcgatca
      961 gatggctccc gcccacgagc ttcagggcca tgtcgtttct gccttccagg ccgccgtcag
     1021 cggggtacag cgtctcggtg aaggcctccc agccgagtgt tttcttctgc atcacagggc
     1081 cgttggatgt gaagttcacc cctctgatct tgacgttgta gatgaggcag ccgtcctgga
     1141 ggctggtgtc ctgggtagcg gtcagcacgc ccccgtcttc gtatgtggtg actctctccc
     1201 atgtgaagcc ctcagggaag gactgcttga agaagtcggg gatgccctgg gtgtggttga
     1261 tgaaggtctt gctgccgtag aggaagctag tagccaggat gtcgaaggcg aaggggagag
     1321 ggccgccctc gaccaccttg attctcatgg tctgggtgcc ctcgtagggc ttgccttcgc
     1381 cctcggatgt gcacttgaag tgatggttgt ccacggtgcc ctccatgtac agcttcatgt
     1441 gcatgttctc cttaatcagc tcgcttggcc cgggattctc ttcgacatcc cctgcttgtt
     1501 tcaacaggga gaagttagtg gctccgcttc cgttatctcc ggacaaggcc tgctcgggtg
     1561 cgcgcttgtg aaaaatgacc gactgtctct gctggtactg ctgcttgcgc ctcttttttt
     1621 tgtcacactg gcacagcagc agcatcttga aagtggttct gaatgttttg ttgcacagag
     1681 catagcacac ggggttcacg gtgctgttga tgtagcacag ccagtagccc agattccaaa
     1741 aggttttggg tatgcagctg tcacaaaagg tgttcaccag aaccatgatg ttgtatgggg
     1801 tccaagtgat gatgaaggca agcaagatcg cactgagggt ctgggccgct ttcttctcct
     1861 tgaccaggga catccttttc cgcttagtga tctgacttct ggtcttcaga gcaaacctct
     1921 tggccagagt ggcttccttg aaggacagag gtagagtggc cgtgctctta cccactgagg
     1981 agttgacgtc agaagtctta gctgtgtcca cggctgactc tagctggatg ggaagcttgg
     2041 agaagctttt tggaaaactg cctccatcgt ccacgctctt ctgggcctgc agcttgtcgg
     2101 ctttcctctc caagtccacc atccccagct cctcctcagg cacctgcagg ttgtccgatg
     2161 agggtaactt ggtggagttg aggatggtgc tgtgacccgg aagcttgagc acgatggagt
     2221 agatggctct cgtctcggag ccaatgtcct cctcgtcgga ggaggcggag ttctccaggg
     2281 aggcagcagc atcattgttg ttccaactgt cactgctgct gtggtcttgg tccatctgct
     2341 cggagctggg tttccagctc ttggttgtga accagaagtg gcagcggcca tacttcctcc
     2401 tgttggagcg tttcatgctt tgctgttgaa gttcgtaact gctgcagctt cgagaactgc
     2461 ccgtggggtg gacaaagttt tctgtctctg cctctgtccc agaggcttgc aggccagcaa
     2521 gctctttggt acgcttttca gtttccttat agatcctcca gtataaaata gtcataatgg
     2581 tgacaggcat ataaaaacca gcgatggctg tgccaaaagt aatggtgggc tcactgagga
     2641 actgaatgaa gcactctccc ggaggcacag ttctctttcc aacaaagtat tgccagaaca
     2701 agatggcagg agcccaaagg acaaaggaga tgacccaagc cagaccgatc atcacaccgg
     2761 ctctctttgt tgttcgtttg gctcggtacg tgagcggcct cgtgatggaa aagtatctgt
     2821 caaagctgat gaccagaaga ttcataacag aggcattgct ggctacgcag tcaatggcaa
     2881 gccagaggtc acaggccaag ttccctaagg cccatcgatt catgatgatg taggtcgtaa
     2941 acagattcat tgaaatgacc ccgataatca gatcggcaca ggccaggctt aagaggaagt
     3001 agttgttgac cgtcttcagc tgcttgttga ccttaaatga cacaattacc aggatgttgc
     3061 cgatgatggt caccaaggcc aggatgcccg ttaagaaagc gatgaagacc acttgccaga
     3121 cggtatgacc tcccagaggg tcatcggtgg taccgtctgg agaggagaaa ttgccagctg
     3181 ctcgagaaac attgtagctg ccgaaatgag tgacggttcc cgggggcagc cctgcatcgg
     3241 aggggctgtg tatccaggag gagctgatgt ttggaaacaa aggcgaggtt gtactgttat
     3301 tgtgcaaggt agcgtaatct ggaacatcgt atgggtacat ggtggcaagc tttactagtg
     3361 gtggcgctag cataacttcg tataaagtat cctatacgaa gttatttgcc ttaacccaga
     3421 aattatcact gttattcttt agaatggtgc aaagaataac ttcgtataat gtatgctata
     3481 cgaagttatg gcgcgccaat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     3541 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     3601 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     3661 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     3721 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     3781 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     3841 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt
     3901 cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     3961 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     4021 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     4081 ccccgcctgg atcctgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     4141 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     4201 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     4261 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggca
     4321 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     4381 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg
     4441 agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     4501 tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc
     4561 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc
     4621 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     4681 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     4741 aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     4801 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     4861 ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat
     4921 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     4981 tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     5041 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     5101 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     5161 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     5221 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     5281 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     5341 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     5401 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     5461 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     5521 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     5581 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     5641 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     5701 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     5761 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     5821 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     5881 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     5941 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     6001 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     6061 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     6121 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     6181 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     6241 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     6301 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     6361 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     6421 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     6481 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     6541 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     6601 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     6661 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     6721 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     6781 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     6841 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     6901 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     6961 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     7021 cggttcctgg ccttttgctg gccttttgct cacatgt
//
