LOCUS       sequence-421887-        6219 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV construct expressing dTomato driven by chandelier cell-targeting
            enhancer. Alias: pAAV_WDC0004_dTomato_nlsdTomato.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6219)
  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 6219)
  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_BiPVe4_dTomato_nlsdTomato
FEATURES             Location/Qualifiers
     source          1..6219
                     /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          838..970
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     regulatory      993..1002
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             999..1700
                     /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             1722..1778
                     /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             1791..1811
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1812..2510
                     /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    2565..3153
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2618..2638)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3036..3047)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3322..3403
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3359..3378)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3413..3432
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   3482..3622
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3482..3611
                     /label=AAV2 ITR
     rep_origin      3697..4152
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3784..3803)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3994..4015
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4169..4188)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4288..4310
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4348..4366)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4434..4538
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4539..4607,4608..5399)
                     /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(4757..4776)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5570..6158
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6059..6078
                     /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 ggtgtatttc ctgtgtgggc taatgtagct
      181 aactttgttt ggactccagg tttctgaggc agtttcaaat tcttgattta tgggttcatt
      241 tttggatcat ggcagcctcc ctgctgaaac ccccatagtt tcccttcaat aaggaaggct
      301 ggatgagatc acagctgagg aatcctggtg gtgtgtactg tgttatttct ctcctagtgc
      361 tttgcctaca cccttccatc atgcttacat ttggacctga aaataattgg ttgaagctgc
      421 tcctcacttt tagcctttta tccgagtttg acagtacatc accccaaata acacggacta
      481 taaatggctt gaacctaatg agtgctcttt gtggacaagc cgtccattct ccttttcttg
      541 agagggaaat taccggctcc taagcttcta tttccagttc tatagaatgt gggcgctaat
      601 ttgaaagcag cctcataaaa tggaagcaaa caggctaatt agaaatattt atgggcctgt
      661 tagtgaatat atgccatcga tgtttaaact tcctgcagcc cgggctgggc ataaaagtca
      721 gggcagagcc atctattgct tacatttgct tctagcctgc aagtcgagga gcgcagcctt
      781 ccagaagcag agcgcggcgc cttaagctgc agaagttggt cgtgaggcac tgggcacgta
      841 agtatcaagg ttacaagaca ggtttaagga gaccaataga aactgggctt gtcgagacag
      901 agaagactct tgcgtttctg ataggcacct attggtctta ctgacatcca ctttgccttt
      961 ctctccacag cgagctcgct agcgcgatcg ctgccaccat ggtgagcaag ggcgaggagg
     1021 tcatcaaaga gttcatgcgc ttcaaggtgc gcatggaggg ctccatgaac ggccacgagt
     1081 tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga
     1141 aggtgaccaa gggcggcccc ctgcccttcg cctgggacat cctgtccccc cagttcatgt
     1201 acggctccaa ggcgtacgtg aagcaccccg ccgacatccc cgattacaag aagctgtcct
     1261 tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggt ctggtgaccg
     1321 tgacccagga ctcctccctg caggacggca cgctgatcta caaggtgaag atgcgcggca
     1381 ccaacttccc ccccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcca
     1441 ccgagcgcct gtacccccgc gacggcgtgc tgaagggcga gatccaccag gccctgaagc
     1501 tgaaggacgg cggccactac ctggtggagt tcaagaccat ctacatggcc aagaagcccg
     1561 tgcaactgcc cggctactac tacgtggaca ccaagctgga catcacctcc cacaacgagg
     1621 actacaccat cgtggaacag tacgagcgct ccgagggccg ccaccacctg ttcctgtacg
     1681 gcatggacga gctgtacaag tccggagata acggaagcgg agccactaac ttctccctgt
     1741 tgaaacaagc aggggatgtc gaagagaatc ccgggccatc tagaatggcc cccaagaaga
     1801 agaggaaggt ggtgagcaag ggcgaggagg tcatcaaaga gttcatgcgc ttcaaggtgc
     1861 gcatggaggg ctccatgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
     1921 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggcggcccc ctgcccttcg
     1981 cctgggacat cctgtccccc cagttcatgt acggctccaa ggcgtacgtg aagcaccccg
     2041 ccgacatccc cgattacaag aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga
     2101 tgaacttcga ggacggcggt ctggtgaccg tgacccagga ctcctccctg caggacggca
     2161 cgctgatcta caaggtgaag atgcgcggca ccaacttccc ccccgacggc cccgtaatgc
     2221 agaagaagac catgggctgg gaggcctcca ccgagcgcct gtacccccgc gacggcgtgc
     2281 tgaagggcga gatccaccag gccctgaagc tgaaggacgg cggccactac ctggtggagt
     2341 tcaagaccat ctacatggcc aagaagcccg tgcaactgcc cggctactac tacgtggaca
     2401 ccaagctgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgagcgct
     2461 ccgagggccg ccaccacctg ttcctgtacg gcatggacga gctgtacaag taagtcgacg
     2521 gcgcgcccca cccctgcagg gaattcgata tcaagcttat cgataatcaa cctctggatt
     2581 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     2641 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     2701 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     2761 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     2821 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     2881 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     2941 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctat gttgccacct
     3001 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     3061 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     3121 cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgagcgctg ctcgagagat
     3181 ctgtgatagc ggccatcaag ctggccgcga ctctagatca taatcagcca taccacattt
     3241 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     3301 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     3361 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     3421 tccaaactca tcaatgtatc agcttatcga taccgcatgc acgtgcggac cgagcggccg
     3481 caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     3541 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag
     3601 cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc
     3661 ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa
     3721 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc
     3781 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     3841 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     3901 aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     3961 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     4021 cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct
     4081 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     4141 cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     4201 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     4261 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     4321 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     4381 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     4441 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     4501 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     4561 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     4621 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     4681 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     4741 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     4801 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     4861 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     4921 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     4981 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     5041 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     5101 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     5161 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     5221 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     5281 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     5341 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     5401 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     5461 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     5521 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     5581 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     5641 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     5701 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     5761 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     5821 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     5881 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     5941 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     6001 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     6061 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     6121 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     6181 tacggttcct ggccttttgc tggccttttg ctcacatgt
//
