LOCUS       sequence-425681-        6092 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV construct expressing dTomato driven by VIP interneuron-targeting
            enhancer. Alias: pAAV_YWE4.2_dTomato_nlsdTomato.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6092)
  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 6092)
  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_BiVIPe4_dTomato_nlsdTomato
FEATURES             Location/Qualifiers
     source          1..6092
                     /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          712..844
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     regulatory      867..876
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             873..1574
                     /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             1596..1652
                     /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             1665..1685
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1686..2384
                     /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    2439..3027
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2492..2512)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2910..2921)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3196..3277
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3233..3252)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3287..3306
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   3356..3496
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3356..3485
                     /label=AAV2 ITR
     rep_origin      3571..4026
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3658..3677)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3868..3889
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4043..4062)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4162..4184
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4222..4240)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4308..4412
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4413..4481,4482..5273)
                     /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(4631..4650)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5444..6032
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5933..5952
                     /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 tattttaaat cctcattggt caggaatgtt
      181 tacattcata gtgtgaagca gagcctgaaa tgccaactta gtacttcagt gtgctgctaa
      241 attgactaag cgtgctgaac agtaatagct tggtactaga agcccaatgg cagggctcag
      301 tggccaaatg tttttattat tacgaagtaa taaggctaat aagcagtgtt gtgtgatgga
      361 aggcaggctt acttctattt aaagatggaa tgaacagctt tgatggggac aagcaaagaa
      421 cacaagaaca ttagctgtgt tccgatattc agtgcacata ggccagagct tggcaaacat
      481 ttctgtaatg gccagatgat aaacatttga ggattggaag actatagggt ctccaatgca
      541 accactaaaa tcgatgttta aacttcctgc agcccgggct gggcataaaa gtcagggcag
      601 agccatctat tgcttacatt tgcttctagc ctgcaagtcg aggagcgcag ccttccagaa
      661 gcagagcgcg gcgccttaag ctgcagaagt tggtcgtgag gcactgggca cgtaagtatc
      721 aaggttacaa gacaggttta aggagaccaa tagaaactgg gcttgtcgag acagagaaga
      781 ctcttgcgtt tctgataggc acctattggt cttactgaca tccactttgc ctttctctcc
      841 acagcgagct cgctagcgcg atcgctgcca ccatggtgag caagggcgag gaggtcatca
      901 aagagttcat gcgcttcaag gtgcgcatgg agggctccat gaacggccac gagttcgaga
      961 tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga
     1021 ccaagggcgg ccccctgccc ttcgcctggg acatcctgtc cccccagttc atgtacggct
     1081 ccaaggcgta cgtgaagcac cccgccgaca tccccgatta caagaagctg tccttccccg
     1141 agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggtctggtg accgtgaccc
     1201 aggactcctc cctgcaggac ggcacgctga tctacaaggt gaagatgcgc ggcaccaact
     1261 tcccccccga cggccccgta atgcagaaga agaccatggg ctgggaggcc tccaccgagc
     1321 gcctgtaccc ccgcgacggc gtgctgaagg gcgagatcca ccaggccctg aagctgaagg
     1381 acggcggcca ctacctggtg gagttcaaga ccatctacat ggccaagaag cccgtgcaac
     1441 tgcccggcta ctactacgtg gacaccaagc tggacatcac ctcccacaac gaggactaca
     1501 ccatcgtgga acagtacgag cgctccgagg gccgccacca cctgttcctg tacggcatgg
     1561 acgagctgta caagtccgga gataacggaa gcggagccac taacttctcc ctgttgaaac
     1621 aagcagggga tgtcgaagag aatcccgggc catctagaat ggcccccaag aagaagagga
     1681 aggtggtgag caagggcgag gaggtcatca aagagttcat gcgcttcaag gtgcgcatgg
     1741 agggctccat gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg
     1801 agggcaccca gaccgccaag ctgaaggtga ccaagggcgg ccccctgccc ttcgcctggg
     1861 acatcctgtc cccccagttc atgtacggct ccaaggcgta cgtgaagcac cccgccgaca
     1921 tccccgatta caagaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact
     1981 tcgaggacgg cggtctggtg accgtgaccc aggactcctc cctgcaggac ggcacgctga
     2041 tctacaaggt gaagatgcgc ggcaccaact tcccccccga cggccccgta atgcagaaga
     2101 agaccatggg ctgggaggcc tccaccgagc gcctgtaccc ccgcgacggc gtgctgaagg
     2161 gcgagatcca ccaggccctg aagctgaagg acggcggcca ctacctggtg gagttcaaga
     2221 ccatctacat ggccaagaag cccgtgcaac tgcccggcta ctactacgtg gacaccaagc
     2281 tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgag cgctccgagg
     2341 gccgccacca cctgttcctg tacggcatgg acgagctgta caagtaagtc gacggcgcgc
     2401 cccacccctg cagggaattc gatatcaagc ttatcgataa tcaacctctg gattacaaaa
     2461 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     2521 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     2581 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     2641 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     2701 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     2761 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     2821 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc acctggattc
     2881 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     2941 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
     3001 ggatctccct ttgggccgcc tccccgcatc gataccgagc gctgctcgag agatctgtga
     3061 tagcggccat caagctggcc gcgactctag atcataatca gccataccac atttgtagag
     3121 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     3181 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     3241 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     3301 ctcatcaatg tatcagctta tcgataccgc atgcacgtgc ggaccgagcg gccgcaggaa
     3361 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg
     3421 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg
     3481 cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     3541 tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg
     3601 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc
     3661 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3721 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     3781 ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     3841 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3901 actctatctc gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt
     3961 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     4021 caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     4081 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     4141 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     4201 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     4261 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     4321 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     4381 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     4441 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     4501 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     4561 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     4621 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     4681 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     4741 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     4801 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     4861 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     4921 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     4981 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     5041 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     5101 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     5161 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     5221 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     5281 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     5341 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     5401 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     5461 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     5521 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     5581 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     5641 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     5701 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     5761 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     5821 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     5881 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     5941 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     6001 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     6061 tcctggcctt ttgctggcct tttctcacat gt
//
