LOCUS       sequence-421889-        6452 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV construct expressing dTomato driven by SST interneuron-targeting
            enhancer. Alias: pAAV_S9E10_dTomato_nlsdTomato.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6452)
  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 6452)
  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_BiSSTe10_dTomato_nlsdTomato
FEATURES             Location/Qualifiers
     source          1..6452
                     /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          1071..1203
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     regulatory      1226..1235
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1232..1933
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     CDS             1955..2011
                     /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             2024..2044
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2045..2743
                     /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"
     misc_feature    2798..3386
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2851..2871)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3269..3280)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3555..3636
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3592..3611)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3646..3665
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   3715..3855
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3715..3844
                     /label=AAV2 ITR
     rep_origin      3930..4385
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4017..4036)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4227..4248
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4402..4421)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4521..4543
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4581..4599)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4667..4771
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4772..4840,4841..5632)
                     /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(4990..5009)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5803..6391
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6292..6311
                     /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 ctaggttcct gtctgcaagc atagcagggt
      181 atcattaata gtgtcaggag ttggctctct cccataggat gggtctcaag ttggggcaat
      241 cattggttgg ctgatccctt ggtctctgtc ccatctttat tcctgcacat cttgtaggca
      301 ggacaaattt tggtttgttc caaagtgatt tttaaagcat taccattttg tgttttgttg
      361 gtttaaaaca ctttagctat tctggtgggt gcgtgatcct gcctcactac ggtctcagtc
      421 tccatctcga tggtgactaa ggatgcctcc tatgcactta tttaccatct gaatggttcc
      481 tttatagaat gcatgactat tcaatttttt tgctcattgg acagcatttt aatgtgtgaa
      541 aggattatac atccttcaag atatggaaga aaaagcacaa agccctaaat tctgtgaatt
      601 cagatgtgta aaaaaagaca aggaagtgca aactatatca gcttggtgtc tcattctggg
      661 tggctgtcag agccctgtga gctgacttaa gaattgggtc cctgcaatgg gtgctggcac
      721 ttgcaagggg atcatgagct caaggccata ctgagctgca taatgagacc ttgtctcagt
      781 aactagactg gggttggagg tcagtactaa agggctagcc tagcctaaat tcccgagttc
      841 cacccccagc actataaaag gaaaaggagg aagaggtgga gggaaataaa gaaaaggaaa
      901 gccagtgtat cgatgtttaa acttcctgca gcccgggctg ggcataaaag tcagggcaga
      961 gccatctatt gcttacattt gcttctagcc tgcaagtcga ggagcgcagc cttccagaag
     1021 cagagcgcgg cgccttaagc tgcagaagtt ggtcgtgagg cactgggcac gtaagtatca
     1081 aggttacaag acaggtttaa ggagaccaat agaaactggg cttgtcgaga cagagaagac
     1141 tcttgcgttt ctgataggca cctattggtc ttactgacat ccactttgcc tttctctcca
     1201 cagcgagctc gctagcgcga tcgctgccac catggtgagc aagggcgagg aggtcatcaa
     1261 agagttcatg cgcttcaagg tgcgcatgga gggctccatg aacggccacg agttcgagat
     1321 cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac
     1381 caagggcggc cccctgccct tcgcctggga catcctgtcc ccccagttca tgtacggctc
     1441 caaggcgtac gtgaagcacc ccgccgacat ccccgattac aagaagctgt ccttccccga
     1501 gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggtctggtga ccgtgaccca
     1561 ggactcctcc ctgcaggacg gcacgctgat ctacaaggtg aagatgcgcg gcaccaactt
     1621 cccccccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct ccaccgagcg
     1681 cctgtacccc cgcgacggcg tgctgaaggg cgagatccac caggccctga agctgaagga
     1741 cggcggccac tacctggtgg agttcaagac catctacatg gccaagaagc ccgtgcaact
     1801 gcccggctac tactacgtgg acaccaagct ggacatcacc tcccacaacg aggactacac
     1861 catcgtggaa cagtacgagc gctccgaggg ccgccaccac ctgttcctgt acggcatgga
     1921 cgagctgtac aagtccggag ataacggaag cggagccact aacttctccc tgttgaaaca
     1981 agcaggggat gtcgaagaga atcccgggcc atctagaatg gcccccaaga agaagaggaa
     2041 ggtggtgagc aagggcgagg aggtcatcaa agagttcatg cgcttcaagg tgcgcatgga
     2101 gggctccatg aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga
     2161 gggcacccag accgccaagc tgaaggtgac caagggcggc cccctgccct tcgcctggga
     2221 catcctgtcc ccccagttca tgtacggctc caaggcgtac gtgaagcacc ccgccgacat
     2281 ccccgattac aagaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt
     2341 cgaggacggc ggtctggtga ccgtgaccca ggactcctcc ctgcaggacg gcacgctgat
     2401 ctacaaggtg aagatgcgcg gcaccaactt cccccccgac ggccccgtaa tgcagaagaa
     2461 gaccatgggc tgggaggcct ccaccgagcg cctgtacccc cgcgacggcg tgctgaaggg
     2521 cgagatccac caggccctga agctgaagga cggcggccac tacctggtgg agttcaagac
     2581 catctacatg gccaagaagc ccgtgcaact gcccggctac tactacgtgg acaccaagct
     2641 ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgagc gctccgaggg
     2701 ccgccaccac ctgttcctgt acggcatgga cgagctgtac aagtaagtcg acggcgcgcc
     2761 ccacccctgc agggaattcg atatcaagct tatcgataat caacctctgg attacaaaat
     2821 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     2881 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     2941 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     3001 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     3061 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     3121 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     3181 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tatgttgcca cctggattct
     3241 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     3301 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     3361 gatctccctt tgggccgcct ccccgcatcg ataccgagcg ctgctcgaga gatctgtgat
     3421 agcggccatc aagctggccg cgactctaga tcataatcag ccataccaca tttgtagagg
     3481 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     3541 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     3601 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     3661 tcatcaatgt atcagcttat cgataccgca tgcacgtgcg gaccgagcgg ccgcaggaac
     3721 ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc
     3781 gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc
     3841 gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     3901 cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc
     3961 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc
     4021 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     4081 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     4141 tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     4201 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     4261 ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg tctattggtt
     4321 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac
     4381 aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     4441 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     4501 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     4561 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     4621 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     4681 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     4741 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     4801 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     4861 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     4921 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     4981 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     5041 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     5101 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     5161 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     5221 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     5281 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     5341 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     5401 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     5461 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     5521 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     5581 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     5641 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     5701 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     5761 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5821 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     5881 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5941 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     6001 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     6061 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     6121 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6181 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6241 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6301 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6361 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     6421 cctggccttt tgctggcctt ttgctcacat gt
//
