LOCUS       sequence-431893-        6323 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV construct with dTomato driven by Lamp5 interneuron-targeting 
            enhancer. Alias: pAAV_WDL0003_dTomato_nlsdTomato.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6323)
  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 6323)
  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_BiLAMP5e3_dTomato_nlsdTomato
FEATURES             Location/Qualifiers
     source          1..6323
                     /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          942..1074
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     regulatory      1097..1106
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1103..1804
                     /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             1826..1882
                     /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             1895..1915
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1916..2614
                     /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    2669..3257
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2722..2742)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3140..3151)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3426..3507
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3463..3482)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3517..3536
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   3586..3726
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3586..3715
                     /label=AAV2 ITR
     rep_origin      3801..4256
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3888..3907)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4098..4119
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4273..4292)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4392..4414
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4452..4470)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4538..4642
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4643..4711,4712..5503)
                     /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(4861..4880)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5674..6262
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6163..6182
                     /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 aaaggatctt ggccctgtac gcctaagcat
      181 cccttcggtt tggtagcaat ggatgattgt atgcaccatt ctggttcaca tgagttttct
      241 ttctttcttt tgacagcatc tgattatatt agtgtgtatt catcaggaac aaaatggttc
      301 catcattatt caagtgactt ggattcagaa tcactagacc tatcgaatat gcagatgcaa
      361 aggaaagttg gtctattcag cctatcccgc aacaagccaa gaaaaccctg gcaccaatca
      421 ggaacaacaa aaatggaaga aagtatgtgc ttaataatcg gtatcattct ctgaagtaaa
      481 taaaagtgct cagattagtg gcaaagctca ttggttcttt taatatcaat tgtaatgcca
      541 tgaatttttc ccttagctac tgtttcttat tgaaatgaac tgaaataact ctaaaagaaa
      601 catctattat atgactgcaa atgttggagc agatataaat ctgaataggc ttatgagtag
      661 agtttctatc tttgccactg gcttttaatt attttcctaa atcacatata actttgatga
      721 aagtgaatat tgctattttc tgtcagccat ggttgaatga catcaacaca ggagccatta
      781 tcgatgttta aacttcctgc agcccgggct gggcataaaa gtcagggcag agccatctat
      841 tgcttacatt tgcttctagc ctgcaagtcg aggagcgcag ccttccagaa gcagagcgcg
      901 gcgccttaag ctgcagaagt tggtcgtgag gcactgggca cgtaagtatc aaggttacaa
      961 gacaggttta aggagaccaa tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt
     1021 tctgataggc acctattggt cttactgaca tccactttgc ctttctctcc acagcgagct
     1081 cgctagcgcg atcgctgcca ccatggtgag caagggcgag gaggtcatca aagagttcat
     1141 gcgcttcaag gtgcgcatgg agggctccat gaacggccac gagttcgaga tcgagggcga
     1201 gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggcgg
     1261 ccccctgccc ttcgcctggg acatcctgtc cccccagttc atgtacggct ccaaggcgta
     1321 cgtgaagcac cccgccgaca tccccgatta caagaagctg tccttccccg agggcttcaa
     1381 gtgggagcgc gtgatgaact tcgaggacgg cggtctggtg accgtgaccc aggactcctc
     1441 cctgcaggac ggcacgctga tctacaaggt gaagatgcgc ggcaccaact tcccccccga
     1501 cggccccgta atgcagaaga agaccatggg ctgggaggcc tccaccgagc gcctgtaccc
     1561 ccgcgacggc gtgctgaagg gcgagatcca ccaggccctg aagctgaagg acggcggcca
     1621 ctacctggtg gagttcaaga ccatctacat ggccaagaag cccgtgcaac tgcccggcta
     1681 ctactacgtg gacaccaagc tggacatcac ctcccacaac gaggactaca ccatcgtgga
     1741 acagtacgag cgctccgagg gccgccacca cctgttcctg tacggcatgg acgagctgta
     1801 caagtccgga gataacggaa gcggagccac taacttctcc ctgttgaaac aagcagggga
     1861 tgtcgaagag aatcccgggc catctagaat ggcccccaag aagaagagga aggtggtgag
     1921 caagggcgag gaggtcatca aagagttcat gcgcttcaag gtgcgcatgg agggctccat
     1981 gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca
     2041 gaccgccaag ctgaaggtga ccaagggcgg ccccctgccc ttcgcctggg acatcctgtc
     2101 cccccagttc atgtacggct ccaaggcgta cgtgaagcac cccgccgaca tccccgatta
     2161 caagaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg
     2221 cggtctggtg accgtgaccc aggactcctc cctgcaggac ggcacgctga tctacaaggt
     2281 gaagatgcgc ggcaccaact tcccccccga cggccccgta atgcagaaga agaccatggg
     2341 ctgggaggcc tccaccgagc gcctgtaccc ccgcgacggc gtgctgaagg gcgagatcca
     2401 ccaggccctg aagctgaagg acggcggcca ctacctggtg gagttcaaga ccatctacat
     2461 ggccaagaag cccgtgcaac tgcccggcta ctactacgtg gacaccaagc tggacatcac
     2521 ctcccacaac gaggactaca ccatcgtgga acagtacgag cgctccgagg gccgccacca
     2581 cctgttcctg tacggcatgg acgagctgta caagtaagtc gacggcgcgc cccacccctg
     2641 cagggaattc gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag
     2701 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     2761 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     2821 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     2881 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     2941 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     3001 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     3061 gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc acctggattc tgcgcgggac
     3121 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     3181 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     3241 ttgggccgcc tccccgcatc gataccgagc gctgctcgag agatctgtga tagcggccat
     3301 caagctggcc gcgactctag atcataatca gccataccac atttgtagag gttttacttg
     3361 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     3421 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     3481 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     3541 tatcagctta tcgataccgc atgcacgtgc ggaccgagcg gccgcaggaa cccctagtga
     3601 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     3661 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     3721 tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     3781 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     3841 ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt
     3901 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     3961 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     4021 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     4081 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc
     4141 gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga
     4201 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     4261 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     4321 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     4381 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     4441 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     4501 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     4561 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4621 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     4681 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     4741 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     4801 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     4861 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     4921 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     4981 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     5041 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     5101 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     5161 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     5221 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     5281 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     5341 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     5401 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     5461 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     5521 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     5581 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     5641 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5701 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5761 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5821 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5881 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5941 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     6001 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     6061 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     6121 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6181 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6241 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     6301 ttgctggcct tttgctcaca tgt
//
