LOCUS       sequence-421867-        7617 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV construct expressing HA-hM3Dq-dTomato driven by cholinergic 
            neuron-targeting enhancer. Alias: pAAV_S9E27_Gq_dTomato.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7617)
  AUTHORS   Furlanis E, Dai M, Leyva Garcia B, Tran T, Vergara J, Pereira A, 
            Gorissen BL, Wills S, Vlachos A, Hairston A, Dwivedi D, Du S, 
            McMahon J, Huang S, Morabito A, Vazquez A, Kim S, Lee AT, Chang EF, 
            Razzaq T, Qazi A, Vargish G, Yuan X, Caccavano A, Hunt S, 
            Chittajallu R, McLean N, Hewitt L, Paranzino E, Rice H, Cummins AC, 
            Plotnikova A, Mohanty A, Tangen AC, Shin JH, Azadi R, Eldridge MAG, 
            Alvarez VA, Averbeck BB, Alyahyay M, Vallejo TR, Soheib M, Vattino 
            LG, MacGregor CP, Chatain CP, Banks E, Olah VJ, Naskar S, Hill S, 
            Liebergall S, Badiani R, Hyde L, Hanley E, Xu Q, Allaway KC, 
            Goldberg EM, Rowan MJM, Nowakowski TJ, Lee S, Favuzzi E, Kaeser PS, 
            Sjulson L, Batista-Brito R, Takesian AE, Ibrahim LA, Iqbal A, Pelkey
            KA, McBain CJ, Dimidschstein J, Fishell G, Wang Y
  TITLE     An enhancer-AAV toolbox to target and manipulate distinct 
            interneuron subtypes.
  JOURNAL   Neuron. 2025 May 21;113(10):1525-1547.e15. doi: 
            10.1016/j.neuron.2025.05.002.
  PUBMED    40403705
REFERENCE   2  (bases 1 to 7617)
  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"; doi: 
            "10.1016/j.neuron.2025.05"; journalName: "Neuron"; date: 
            "2025-05-21- 21"; volume: "113"; issue: "10"; pages: "1525-1547.e15"
COMMENT     Sequence Label: pAAV_BiCHATe27_Gq_dTomato
FEATURES             Location/Qualifiers
     source          1..7617
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     intron          1150..1282
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     CDS             1305..1331
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3120..3176
                     /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             3189..3209
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3210..3908
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGL
                     VTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIHQ
                     ALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRHH
                     LFLYGMDELYK"
     primer_bind     complement(3340..3358)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3608..3627
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3802..3825
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     misc_feature    3963..4551
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4016..4036)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4434..4445)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4720..4801
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4757..4776)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4811..4830
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   4880..5020
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4880..5009
                     /label=AAV2 ITR
     rep_origin      5095..5550
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5182..5201)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5392..5413
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5567..5586)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5686..5708
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        5832..5936
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5937..6005,6006..6797)
                     /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(6155..6174)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6968..7556
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7457..7476
                     /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 gcgtatcgat cctcgggtaa tttgtaaaga caatttttct
      181 tttccccctc tgaacctagg ccttaggctg gctaggcaag tctccactac agccccgccc
      241 actccatctc ggctgtggct cgcgggaaag gcagccccct acagttagtc tccccagcca
      301 tcttctccgg ttcccctgat gctctcatgg ccccagccat cctagagctg gtgcgccatc
      361 gctcaccttt agcattcggc ctgctggggg tgagcttttg ggttagaagg aacaaggcat
      421 cagatggggt cgtgtttttt tacattccaa tcagttttcg gaggggaaga acagacattt
      481 ttgtcagcta attgcattta attacagaga caatcgctgt ttccgaacgt tgttgtgtcc
      541 actccctcat ccttcaatca gttcaagcac atccggcccc agctgggagc cttttaattg
      601 cataatttgg atttcacttc tggaaagaaa cttgagaaag actccgtgag ggtctccatt
      661 aacacaatca gctccccagg cacccggggc tggccctaat tgacccatct cagctaatgc
      721 ctctttccaa agaacgcccg cgtcatttct ttggcttgtt tgtgctattg tatcctgttt
      781 tagtgtcttg atcttaaaca tctttaaaga gggcactcag tggcttgttt aactggaagt
      841 ggtccagctg gtttggaagg acagtgcatc aggtgatgta agtccaaggg taggaatttc
      901 tggcctctcc ttcccctgcc cattgcaccg ctctccccaa caggaagtag ctacacatga
      961 taaggtgcac tgtggcttct aggacttatc gatgtttaaa cttcctgcag cccgggctgg
     1021 gcataaaagt cagggcagag ccatctattg cttacatttg cttctagcct gcaagtcgag
     1081 gagcgcagcc ttccagaagc agagcgcggc gccttaagct gcagaagttg gtcgtgaggc
     1141 actgggcacg taagtatcaa ggttacaaga caggtttaag gagaccaata gaaactgggc
     1201 ttgtcgagac agagaagact cttgcgtttc tgataggcac ctattggtct tactgacatc
     1261 cactttgcct ttctctccac agcgagctcg ctagcgccac catgtaccca tacgatgttc
     1321 cagattacgc taccttgcac aataacagta caacctcgcc tttgtttcca aacatcagct
     1381 cctcctggat acacagcccc tccgatgcag ggctgccccc gggaaccgtc actcatttcg
     1441 gcagctacaa tgtttctcga gcagctggca atttctcctc tccagacggt accaccgatg
     1501 accctctggg aggtcatacc gtctggcaag tggtcttcat cgctttctta acgggcatcc
     1561 tggccttggt gaccatcatc ggcaacatcc tggtaattgt gtcatttaag gtcaacaagc
     1621 agctgaagac ggtcaacaac tacttcctct taagcctggc ctgtgccgat ctgattatcg
     1681 gggtcatttc aatgaatctg tttacgacct acatcatcat gaatcgatgg gccttaggga
     1741 acttggcctg tgacctctgg cttgccattg actgcgtagc cagcaatgcc tctgttatga
     1801 atcttctggt catcagcttt gacagatact tttccatcac gaggccgctc acgtaccgag
     1861 ccaaacgaac aacaaagaga gccggtgtga tgatcggtct ggcttgggtc atctcctttg
     1921 tcctttgggc tcctgccatc ttgttctggc aatactttgt tggaaagaga actgtgcctc
     1981 cgggagagtg cttcattcag ttcctcagtg agcccaccat tacttttggc acagccatcg
     2041 ctggttttta tatgcctgtc accattatga ctattttata ctggaggatc tataaggaaa
     2101 ctgaaaagcg taccaaagag cttgctggcc tgcaagcctc tgggacagag gcagagacag
     2161 aaaactttgt ccaccccacg ggcagttctc gaagctgcag cagttacgaa cttcaacagc
     2221 aaagcatgaa acgctccaac aggaggaagt atggccgctg ccacttctgg ttcacaacca
     2281 agagctggaa acccagctcc gagcagatgg accaagacca cagcagcagt gacagttgga
     2341 acaacaatga tgctgctgcc tccctggaga actccgcctc ctccgacgag gaggacattg
     2401 gctccgagac gagagccatc tactccatcg tgctcaagct tccgggtcac agcaccatcc
     2461 tcaactccac caagttaccc tcatcggaca acctgcaggt gcctgaggag gagctgggga
     2521 tggtggactt ggagaggaaa gccgacaagc tgcaggccca gaagagcgtg gacgatggag
     2581 gcagttttcc aaaaagcttc tccaagcttc ccatccagct agagtcagcc gtggacacag
     2641 ctaagacttc tgacgtcaac tcctcagtgg gtaagagcac ggccactcta cctctgtcct
     2701 tcaaggaagc cactctggcc aagaggtttg ctctgaagac cagaagtcag atcactaagc
     2761 ggaaaaggat gtccctggtc aaggagaaga aagcggccca gaccctcagt gcgatcttgc
     2821 ttgccttcat catcacttgg accccataca acatcatggt tctggtgaac accttttgtg
     2881 acagctgcat acccaaaacc ttttggaatc tgggctactg gctgtgctac atcaacagca
     2941 ccgtgaaccc cgtgtgctat gctctgtgca acaaaacatt cagaaccact ttcaagatgc
     3001 tgctgctgtg ccagtgtgac aaaaaaaaga ggcgcaagca gcagtaccag cagagacagt
     3061 cggtcatttt tcacaagcgc gcacccgagc aggccttgtc cggagataac ggaagcggag
     3121 ccactaactt ctccctgttg aaacaagcag gggatgtcga agagaatccc gggccatcta
     3181 gaatggcccc caagaagaag aggaaggtgg tgagcaaggg cgaggaggtc atcaaagagt
     3241 tcatgcgctt caaggtgcgc atggagggct ccatgaacgg ccacgagttc gagatcgagg
     3301 gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg
     3361 gcggccccct gcccttcgcc tgggacatcc tgtcccccca gttcatgtac ggctccaagg
     3421 cgtacgtgaa gcaccccgcc gacatccccg attacaagaa gctgtccttc cccgagggct
     3481 tcaagtggga gcgcgtgatg aacttcgagg acggcggtct ggtgaccgtg acccaggact
     3541 cctccctgca ggacggcacg ctgatctaca aggtgaagat gcgcggcacc aacttccccc
     3601 ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctccacc gagcgcctgt
     3661 acccccgcga cggcgtgctg aagggcgaga tccaccaggc cctgaagctg aaggacggcg
     3721 gccactacct ggtggagttc aagaccatct acatggccaa gaagcccgtg caactgcccg
     3781 gctactacta cgtggacacc aagctggaca tcacctccca caacgaggac tacaccatcg
     3841 tggaacagta cgagcgctcc gagggccgcc accacctgtt cctgtacggc atggacgagc
     3901 tgtacaagta agtcgacggc gcgccccacc cctgcaggga attcgatatc aagcttatcg
     3961 ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     4021 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     4081 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     4141 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     4201 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     4261 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     4321 tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc
     4381 tcgcctatgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     4441 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     4501 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc
     4561 gagcgctgct cgagagatct gtgatagcgg ccatcaagct ggccgcgact ctagatcata
     4621 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     4681 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     4741 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     4801 cattctagtt gtggtttgtc caaactcatc aatgtatcag cttatcgata ccgcatgcac
     4861 gtgcggaccg agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc
     4921 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg
     4981 gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt
     5041 ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc
     5101 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     5161 ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     5221 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     5281 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     5341 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     5401 tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga
     5461 ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     5521 attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg
     5581 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     5641 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     5701 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     5761 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     5821 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     5881 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     5941 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     6001 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     6061 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     6121 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     6181 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     6241 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     6301 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     6361 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     6421 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     6481 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     6541 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     6601 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     6661 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     6721 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     6781 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     6841 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     6901 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     6961 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     7021 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     7081 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     7141 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     7201 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     7261 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     7321 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     7381 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     7441 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     7501 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     7561 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgt
//
