LOCUS       sequence-425797-        7133 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV construct expressing dTomato driven by Lamp5 
            interneuron-targeting enhancer. Alias: 
            pAAV_WDL0003_dTomato_nlsdTomato_nlsdTomato.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7133)
  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 7133)
  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_nlsdTomato
FEATURES             Location/Qualifiers
     source          1..7133
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   651..780
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     intron          1592..1724
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     regulatory      1747..1756
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1753..2454
                     /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             2476..2532
                     /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             2545..2565
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2566..3264
                     /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"
     CDS             3286..3342
                     /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             3355..3375
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3376..4074
                     /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    4129..4717
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4182..4202)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4600..4611)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4886..4967
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4923..4942)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4977..4996
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   5046..5186
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5046..5175
                     /label=AAV2 ITR
     rep_origin      5261..5716
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5348..5367)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5558..5579
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5733..5752)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5852..5874
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5912..5930)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5998..6102
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6103..6171,6172..6963)
                     /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(6321..6340)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt cctgcaggca
      661 gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt ggtcgcccgg
      721 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
      781 gcggccgcac gcgtatcgat aaaggatctt ggccctgtac gcctaagcat cccttcggtt
      841 tggtagcaat ggatgattgt atgcaccatt ctggttcaca tgagttttct ttctttcttt
      901 tgacagcatc tgattatatt agtgtgtatt catcaggaac aaaatggttc catcattatt
      961 caagtgactt ggattcagaa tcactagacc tatcgaatat gcagatgcaa aggaaagttg
     1021 gtctattcag cctatcccgc aacaagccaa gaaaaccctg gcaccaatca ggaacaacaa
     1081 aaatggaaga aagtatgtgc ttaataatcg gtatcattct ctgaagtaaa taaaagtgct
     1141 cagattagtg gcaaagctca ttggttcttt taatatcaat tgtaatgcca tgaatttttc
     1201 ccttagctac tgtttcttat tgaaatgaac tgaaataact ctaaaagaaa catctattat
     1261 atgactgcaa atgttggagc agatataaat ctgaataggc ttatgagtag agtttctatc
     1321 tttgccactg gcttttaatt attttcctaa atcacatata actttgatga aagtgaatat
     1381 tgctattttc tgtcagccat ggttgaatga catcaacaca ggagccatta tcgatgttta
     1441 aacttcctgc agcccgggct gggcataaaa gtcagggcag agccatctat tgcttacatt
     1501 tgcttctagc ctgcaagtcg aggagcgcag ccttccagaa gcagagcgcg gcgccttaag
     1561 ctgcagaagt tggtcgtgag gcactgggca cgtaagtatc aaggttacaa gacaggttta
     1621 aggagaccaa tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt tctgataggc
     1681 acctattggt cttactgaca tccactttgc ctttctctcc acagcgagct cgctagcgcg
     1741 atcgctgcca ccatggtgag caagggcgag gaggtcatca aagagttcat gcgcttcaag
     1801 gtgcgcatgg agggctccat gaacggccac gagttcgaga tcgagggcga gggcgagggc
     1861 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggcgg ccccctgccc
     1921 ttcgcctggg acatcctgtc cccccagttc atgtacggct ccaaggcgta cgtgaagcac
     1981 cccgccgaca tccccgatta caagaagctg tccttccccg agggcttcaa gtgggagcgc
     2041 gtgatgaact tcgaggacgg cggtctggtg accgtgaccc aggactcctc cctgcaggac
     2101 ggcacgctga tctacaaggt gaagatgcgc ggcaccaact tcccccccga cggccccgta
     2161 atgcagaaga agaccatggg ctgggaggcc tccaccgagc gcctgtaccc ccgcgacggc
     2221 gtgctgaagg gcgagatcca ccaggccctg aagctgaagg acggcggcca ctacctggtg
     2281 gagttcaaga ccatctacat ggccaagaag cccgtgcaac tgcccggcta ctactacgtg
     2341 gacaccaagc tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgag
     2401 cgctccgagg gccgccacca cctgttcctg tacggcatgg acgagctgta caagtccgga
     2461 gataacggaa gcggagccac taacttctcc ctgttgaaac aagcagggga tgtcgaagag
     2521 aatcccgggc catctagaat ggcccccaag aagaagagga aggtggtgag caagggcgag
     2581 gaggtcatca aagagttcat gcgcttcaag gtgcgcatgg agggctccat gaacggccac
     2641 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag
     2701 ctgaaggtga ccaagggcgg ccccctgccc ttcgcctggg acatcctgtc cccccagttc
     2761 atgtacggct ccaaggcgta cgtgaagcac cccgccgaca tccccgatta caagaagctg
     2821 tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggtctggtg
     2881 accgtgaccc aggactcctc cctgcaggac ggcacgctga tctacaaggt gaagatgcgc
     2941 ggcaccaact tcccccccga cggccccgta atgcagaaga agaccatggg ctgggaggcc
     3001 tccaccgagc gcctgtaccc ccgcgacggc gtgctgaagg gcgagatcca ccaggccctg
     3061 aagctgaagg acggcggcca ctacctggtg gagttcaaga ccatctacat ggccaagaag
     3121 cccgtgcaac tgcccggcta ctactacgtg gacaccaagc tggacatcac ctcccacaac
     3181 gaggactaca ccatcgtgga acagtacgag cgctccgagg gccgccacca cctgttcctg
     3241 tacggcatgg acgagctgta caagtccgga gataacggaa gcggagccac taacttctcc
     3301 ctgttgaaac aagcagggga tgtcgaagag aatcccgggc catctagaat ggcccccaag
     3361 aagaagagga aggtggtgag caagggcgag gaggtcatca aagagttcat gcgcttcaag
     3421 gtgcgcatgg agggctccat gaacggccac gagttcgaga tcgagggcga gggcgagggc
     3481 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggcgg ccccctgccc
     3541 ttcgcctggg acatcctgtc cccccagttc atgtacggct ccaaggcgta cgtgaagcac
     3601 cccgccgaca tccccgatta caagaagctg tccttccccg agggcttcaa gtgggagcgc
     3661 gtgatgaact tcgaggacgg cggtctggtg accgtgaccc aggactcctc cctgcaggac
     3721 ggcacgctga tctacaaggt gaagatgcgc ggcaccaact tcccccccga cggccccgta
     3781 atgcagaaga agaccatggg ctgggaggcc tccaccgagc gcctgtaccc ccgcgacggc
     3841 gtgctgaagg gcgagatcca ccaggccctg aagctgaagg acggcggcca ctacctggtg
     3901 gagttcaaga ccatctacat ggccaagaag cccgtgcaac tgcccggcta ctactacgtg
     3961 gacaccaagc tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgag
     4021 cgctccgagg gccgccacca cctgttcctg tacggcatgg acgagctgta caagtaagtc
     4081 gacggcgcgc cccacccctg cagggaattc gatatcaagc ttatcgataa tcaacctctg
     4141 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     4201 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     4261 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     4321 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     4381 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     4441 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     4501 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc
     4561 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     4621 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     4681 cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgagc gctgctcgag
     4741 agatctgtga tagcggccat caagctggcc gcgactctag atcataatca gccataccac
     4801 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     4861 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     4921 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     4981 tttgtccaaa ctcatcaatg tatcagctta tcgataccgc atgcacgtgc ggaccgagcg
     5041 gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
     5101 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag
     5161 cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct
     5221 gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca
     5281 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta
     5341 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     5401 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     5461 cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     5521 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     5581 acaacactca actctatctc gggctattct tttgatttat aagggatttt gccgatttcg
     5641 gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     5701 ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     5761 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     5821 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     5881 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     5941 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     6001 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     6061 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     6121 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     6181 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     6241 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     6301 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     6361 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     6421 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     6481 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     6541 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     6601 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     6661 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     6721 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     6781 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     6841 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     6901 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     6961 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     7021 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     7081 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttc
//
