LOCUS       sequence-244359-        6626 bp    DNA     circular SYN 14-APR-2025
DEFINITION  An adeno-associated viral vector expressing Cre-dependent HA-tagged 
            inhibitory DREADD (hM3D(Gq)) from a synapsin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6626)
  TITLE     AAV for neuronal expression of Cre-dependent DREADDs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6626)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pOTTC1761 - pAAV SYN1 DIO HA-hM3D(Gq)
FEATURES             Location/Qualifiers
     source          1..6626
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        117..564
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(602..635)
                     /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(686..719)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(2498..2524)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    2553..2586
                     /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    2637..2670
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2695..3283
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2748..2768)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3166..3177)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3315..3791
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3514..3533)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3831..3971
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3831..3960
                     /label=AAV2 ITR
     rep_origin      4046..4501
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4133..4152)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4343..4364
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4518..4537)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4637..4659
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        4783..4887
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4888..4956,4957..5748)
                     /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(5106..5125)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5919..6507
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6408..6427
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(6569..6626,1..72)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc aactccatca
       61 ctaggggttc ctgcggccgc acgcgtgtgt ctagactgca gagggccctg cgtatgagtg
      121 caagtgggtt ttaggaccag gatgaggcgg ggtgggggtg cctacctgac gaccgacccc
      181 gacccactgg acaagcaccc aacccccatt ccccaaattg cgcatcccct atcagagagg
      241 gggaggggaa acaggatgcg gcgaggcgcg tgcgcactgc cagcttcagc accgcggaca
      301 gtgccttcgc ccccgcctgg cggcgcgcgc caccgccgcc tcagcactga aggcgcgctg
      361 acgtcactcg ccggtccccc gcaaactccc cttcccggcc accttggtcg cgtccgcgcc
      421 gccgccggcc cagccggacc gcaccacgcg aggcgcgaga taggggggca cgggcgcgac
      481 catctgcgct gcggcgccgg cgactcagcg ctgcctcagt ctgcggtggg cagcggagga
      541 gtcgtgtcgt gcctgagagc gcagtcgaga aggtaccgga tcctctagag tcgactccgg
      601 aataacttcg tataggatac tttatacgaa gttatgcaga atggtagctg gattgtagct
      661 gctattagca atatgaaacc tcttaataac ttcgtatagc atacattata cgaagttatg
      721 gcgcgcccta caaggcctgc tcgggtgcgc gcttgtgaaa aatgaccgac tgtctctgct
      781 ggtactgctg cttgcgcctc ttttttttgt cacactggca cagcagcagc atcttgaaag
      841 tggttctgaa tgttttgttg cacagagcat agcacacggg gttcacggtg ctgttgatgt
      901 agcacagcca gtagcccaga ttccaaaagg ttttgggtat gcagctgtca caaaaggtgt
      961 tcaccagaac catgatgttg tatggggtcc aagtgatgat gaaggcaagc aagatcgcac
     1021 tgagggtctg ggccgctttc ttctccttga ccagggacat ccttttccgc ttagtgatct
     1081 gacttctggt cttcagagca aacctcttgg ccagagtggc ttccttgaag gacagaggta
     1141 gagtggccgt gctcttaccc actgaggagt tgacgtcaga agtcttagct gtgtccacgg
     1201 ctgactctag ctggatggga agcttggaga agctttttgg aaaactgcct ccatcgtcca
     1261 cgctcttctg ggcctgcagc ttgtcggctt tcctctccaa gtccaccatc cccagctcct
     1321 cctcaggcac ctgcaggttg tccgatgagg gtaacttggt ggagttgagg atggtgctgt
     1381 gacccggaag cttgagcacg atggagtaga tggctctcgt ctcggagcca atgtcctcct
     1441 cgtcggagga ggcggagttc tccagggagg cagcagcatc attgttgttc caactgtcac
     1501 tgctgctgtg gtcttggtcc atctgctcgg agctgggttt ccagctcttg gttgtgaacc
     1561 agaagtggca gcggccatac ttcctcctgt tggagcgttt catgctttgc tgttgaagtt
     1621 cgtaactgct gcagcttcga gaactgcccg tggggtggac aaagttttct gtctctgcct
     1681 ctgtcccaga ggcttgcagg ccagcaagct ctttggtacg cttttcagtt tccttataga
     1741 tcctccagta taaaatagtc ataatggtga caggcatata aaaaccagcg atggctgtgc
     1801 caaaagtaat ggtgggctca ctgaggaact gaatgaagca ctctcccgga ggcacagttc
     1861 tctttccaac aaagtattgc cagaacaaga tggcaggagc ccaaaggaca aaggagatga
     1921 cccaagccag accgatcatc acaccggctc tctttgttgt tcgtttggct cggtacgtga
     1981 gcggcctcgt gatggaaaag tatctgtcaa agctgatgac cagaagattc ataacagagg
     2041 cattgctggc tacgcagtca atggcaagcc agaggtcaca ggccaagttc cctaaggccc
     2101 atcgattcat gatgatgtag gtcgtaaaca gattcattga aatgaccccg ataatcagat
     2161 cggcacaggc caggcttaag aggaagtagt tgttgaccgt cttcagctgc ttgttgacct
     2221 taaatgacac aattaccagg atgttgccga tgatggtcac caaggccagg atgcccgtta
     2281 agaaagcgat gaagaccact tgccagacgg tatgacctcc cagagggtca tcggtggtac
     2341 cgtctggaga ggagaaattg ccagctgctc gagaaacatt gtagctgccg aaatgagtga
     2401 cggttcccgg gggcagccct gcatcggagg ggctgtgtat ccaggaggag ctgatgtttg
     2461 gaaacaaagg cgaggttgta ctgttattgt gcaaggtagc gtaatctgga acatcgtatg
     2521 ggtacatggt ggcgtcgact ctagagctag cgataacttc gtataaagta tcctatacga
     2581 agttatttgc cttaacccag aaattatcac tgttattctt tagaatggtg caaagaataa
     2641 cttcgtataa tgtatgctat acgaagttat gaattcgata tcaagcttat cgataatcaa
     2701 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     2761 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     2821 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     2881 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     2941 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     3001 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     3061 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctat
     3121 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     3181 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     3241 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgagcgctg
     3301 ctcgagagat ctacgggtgg catccctgtg acccctcccc agtgcctctc ctggccctgg
     3361 aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc atcattttgt
     3421 ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg agcaaggggc
     3481 aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc tggagtgcag
     3541 tggcacaatc ttggctcact gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc
     3601 agcctcccga gttgttggga ttccaggcat gcatgaccag gctcagctaa tttttgtttt
     3661 tttggtagag acggggtttc accatattgg ccaggctggt ctccaactcc taatctcagg
     3721 tgatctaccc accttggcct cccaaattgc tgggattaca ggcgtgaacc actgctccct
     3781 tccctgtcct tctgattttg taggtaacca cgtgcggacc gagcggccgc aggaacccct
     3841 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     3901 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
     3961 ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     4021 ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt
     4081 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc
     4141 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     4201 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     4261 ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     4321 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct
     4381 atctcgggct attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa
     4441 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
     4501 ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     4561 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     4621 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     4681 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     4741 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     4801 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     4861 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     4921 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     4981 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     5041 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     5101 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     5161 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     5221 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     5281 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     5341 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     5401 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     5461 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     5521 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     5581 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     5641 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     5701 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     5761 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     5821 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     5881 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     5941 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     6001 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     6061 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     6121 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     6181 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     6241 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     6301 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     6361 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     6421 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     6481 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     6541 gccttttgct ggccttttgc tcacatgtcc tgcaggcagc tgcgcgctcg ctcgctcact
     6601 gaggccgccc gggcgtcggg cgacct
//
