LOCUS       sequence-460815-        5430 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV construct expressing eGFP driven by chandelier cell-targeting 
            enhancer. Alias: pAAV_WDC0004_eGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5430)
  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 5430)
  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_eGFP
FEATURES             Location/Qualifiers
     source          1..5430
                     /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             join(999..1001,1002..1004,1005..1718)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(999..1001,1002..1004,1005..1715)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1044..1065)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1305..1324)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1652..1673
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1776..2364
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1829..1849)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2247..2258)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2533..2614
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2570..2589)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2624..2643
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   2693..2833
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2693..2822
                     /label=AAV2 ITR
     rep_origin      2908..3363
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2995..3014)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3205..3226
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3380..3399)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3499..3521
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3559..3577)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3645..3749
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3750..3818,3819..4610)
                     /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(3968..3987)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4781..5369
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5270..5289
                     /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 ggcgaggagc
     1021 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     1081 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
     1141 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     1201 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     1261 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     1321 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     1381 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     1441 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     1501 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     1561 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
     1621 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     1681 ccgggatcac tctcggcatg gacgagctgt acaagtaaga ataagtcgac ggcgcgcccc
     1741 acccctgcag ggaattcgat atcaagctta tcgataatca acctctggat tacaaaattt
     1801 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     1861 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     1921 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     1981 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     2041 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     2101 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     2161 tgtcggggaa atcatcgtcc tttccttggc tgctcgccta tgttgccacc tggattctgc
     2221 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     2281 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     2341 tctccctttg ggccgcctcc ccgcatcgat accgagcgct gctcgagaga tctgtgatag
     2401 cggccatcaa gctggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt
     2461 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     2521 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     2581 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     2641 atcaatgtat cagcttatcg ataccgcatg cacgtgcgga ccgagcggcc gcaggaaccc
     2701 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
     2761 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc
     2821 agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     2881 caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg
     2941 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt
     3001 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     3061 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     3121 atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     3181 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact
     3241 ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa
     3301 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
     3361 ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     3421 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     3481 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     3541 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     3601 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     3661 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     3721 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     3781 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     3841 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     3901 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     3961 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     4021 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     4081 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     4141 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     4201 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     4261 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     4321 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     4381 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     4441 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     4501 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     4561 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     4621 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     4681 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     4741 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     4801 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     4861 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     4921 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     4981 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     5041 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     5101 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     5161 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     5221 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     5281 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     5341 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     5401 tggccttttg ctggcctttt gctcacatgt
//
