LOCUS       sequence-244357-        6296 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An adeno-associated viral vector expressing Cre-dependent HA-tagged 
            inhibitory DREADD (hM4D(Gi)) from a synapsin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6296)
  TITLE     AAV for neuronal expression of Cre-dependent DREADDs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6296)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pOTTC1740 - pAAV SYN1 DIO HA-hM4D(Gi)
FEATURES             Location/Qualifiers
     source          1..6296
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        83..530
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(568..601)
                     /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(652..685)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(2134..2160)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    2189..2222
                     /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    2273..2306
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2331..2919
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2384..2404)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2802..2813)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2951..3427
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3150..3169)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3467..3607
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3467..3596
                     /label=AAV2 ITR
     rep_origin      3682..4137
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3769..3788)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3979..4000
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4154..4173)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4273..4295
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4333..4351)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4419..4523
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4524..4592,4593..5384)
                     /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(4742..4761)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5555..6143
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6044..6063
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(6205..6296,1..38)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 cagagaggga gtggccaact ccatcactag gggttcctgc ggccgcacgc gtgtgtctag
       61 actgcagagg gccctgcgta tgagtgcaag tgggttttag gaccaggatg aggcggggtg
      121 ggggtgccta cctgacgacc gaccccgacc cactggacaa gcacccaacc cccattcccc
      181 aaattgcgca tcccctatca gagaggggga ggggaaacag gatgcggcga ggcgcgtgcg
      241 cactgccagc ttcagcaccg cggacagtgc cttcgccccc gcctggcggc gcgcgccacc
      301 gccgcctcag cactgaaggc gcgctgacgt cactcgccgg tcccccgcaa actccccttc
      361 ccggccacct tggtcgcgtc cgcgccgccg ccggcccagc cggaccgcac cacgcgaggc
      421 gcgagatagg ggggcacggg cgcgaccatc tgcgctgcgg cgccggcgac tcagcgctgc
      481 ctcagtctgc ggtgggcagc ggaggagtcg tgtcgtgcct gagagcgcag tcgagaaggt
      541 accggatcct ctagagtcga ctccggaata acttcgtata ggatacttta tacgaagtta
      601 tgcagaatgg tagctggatt gtagctgcta ttagcaatat gaaacctctt aataacttcg
      661 tatagcatac attatacgaa gttatggcgc gccctacctg gcagtgccga tgttccgata
      721 ctggcacagc agcaggtgcc ggaaggtctt tttaaaggtg gcgttgcaca gagcatagca
      781 ggcagggttg atggtgctgt tgacgtagca gagccagtag ccaatggacc acaccgtgtc
      841 agggatgcag ctctggcaga aggtgttcac caggaccatg acgttgtagg gcgtccaggt
      901 gaggatgaag gcaagcagaa tggcaaagat cgttcgtgtc actttgcgct cccgggccgc
      961 catctgccgc ttcttgcgca cctggttgcg agcgatgctg gcgaacttgc gggccacgtt
     1021 ggccgcaggg cgcatgccag ccggcgtggc aggcacaatc tcaatggctg tcacacactc
     1081 attgcctgtc tgcttcgtca caatctggat cttggaccat ctggaggctg ggttgagggc
     1141 ccgcggctgc aggggagggg cgggcatggc gggcgtggtg gcctctgtgg tggacagctc
     1201 tgtggctggg cgttccttgg tgttctgggt ggcactgcct gagctggact cattggaagt
     1261 gtccttatca gccacggggc gcggtggcgg tggcagcgct ggcggggggg cctcctccag
     1321 cttgccattg cgcagctcct cccgggcggc ctccccgggc gggggcttct tgacgctctg
     1381 cttcattagt gggctcttga ggaaggccag cgtcttggct ttcttctcct tcgggccctc
     1441 gggccggtgc ttgtggactc ggctgcgact ggccagggag atgtggatgt acagcaccgt
     1501 catgatgacc acaggcaggt agaagccagc aatggctgtg ccaaaggtca ctgctgggtt
     1561 ggacaggaac tggatgaagc actggttgtc gggcaccgtc cgcttaccca ccacaaactg
     1621 ccagaacaag atggcaggcg cccagagcac gaaggacagt acccaggcag cagcaatcat
     1681 gaggcctgcc atcttggtgg tgcgccgggc agggtaggtg agaggcttgg tgacgcagaa
     1741 gtagcggtca aagctgatga tgagaaggtt catgacggag gcgttgctca ccacgcagtc
     1801 cagggccagc cacaggtcgc agaccacggc gcccaggggc cagtagccct tgatgatgta
     1861 cacggtgtag aggttcatgg agaaggcgcc tatgatgaga tcagcacacg ccaggctgaa
     1921 gaggaagtag ttgttgactg tctgcagctg cctgttgacc ttgatggaca gcatcaccag
     1981 gatgttgccc acgacagtca ccaggctcag ggagcctgtc actgtggcaa tgaagaccat
     2041 ttccaccgtc tcatagcgat tgtgggatga tgacgtgacc aggcgcacgg actgattgcc
     2101 cgagctgcca ttgactggtg tgaagttggc catagcgtaa tctggaacat cgtatgggta
     2161 catggtggcg tcgactctag agctagcgat aacttcgtat aaagtatcct atacgaagtt
     2221 atttgcctta acccagaaat tatcactgtt attctttaga atggtgcaaa gaataacttc
     2281 gtataatgta tgctatacga agttatgaat tcgatatcaa gcttatcgat aatcaacctc
     2341 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     2401 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     2461 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     2521 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     2581 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     2641 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     2701 acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctatgttg
     2761 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     2821 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     2881 ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccga gcgctgctcg
     2941 agagatctac gggtggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt
     3001 tgccactcca gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga
     3061 ctaggtgtcc ttctataata ttatggggtg gaggggggtg gtatggagca aggggcaagt
     3121 tgggaagaca acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc
     3181 acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc
     3241 tcccgagttg ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg
     3301 gtagagacgg ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat
     3361 ctacccacct tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc
     3421 tgtccttctg attttgtagg taaccacgtg cggaccgagc ggccgcagga acccctagtg
     3481 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag
     3541 gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc
     3601 ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     3661 atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     3721 tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt
     3781 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     3841 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg
     3901 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     3961 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct
     4021 cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg
     4081 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat
     4141 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     4201 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     4261 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     4321 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     4381 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     4441 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     4501 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     4561 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     4621 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     4681 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     4741 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     4801 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     4861 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     4921 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     4981 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     5041 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     5101 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     5161 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     5221 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     5281 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     5341 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     5401 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5461 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5521 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5581 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5641 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     5701 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5761 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5821 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     5881 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     5941 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6001 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6061 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6121 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     6181 tttgctggcc ttttgctcac atgtcctgca ggcagctgcg cgctcgctcg ctcactgagg
     6241 ccgcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg agcgcg
//
