LOCUS       sequence-421870-        6148 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV construct expressing GCaMP6f driven by Lamp5 
            interneuron-targeting enhancer. Alias: pAAV_WDL0003_GCaMP6f.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6148)
  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 6148)
  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_GCaMP6f
FEATURES             Location/Qualifiers
     source          1..6148
                     /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      1095..1104
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1101..1109,1110..1127,1128..1130,1131..1163,1164..1166,
                     1167..1190,1191..1217,1218..1274,1275..1280,1281..1550,
                     1551..1568,1569..2003,2004..2009,2010..2453)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6f
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEEFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYRDTEEEIREAFGVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      1197..1206
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1487..1508
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1614..1635)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1875..1894)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2494..3082
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2547..2567)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2965..2976)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3251..3332
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3288..3307)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3342..3361
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   3411..3551
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3411..3540
                     /label=AAV2 ITR
     rep_origin      3626..4081
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3713..3732)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3923..3944
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4098..4117)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4217..4239
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4277..4295)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4363..4467
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4468..4536,4537..5328)
                     /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(4686..4705)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5499..6087
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5988..6007
                     /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 cgctagcgcc cggtgccacc atgggttctc atcatcatca tcatcatggt atggctagca
     1141 tgactggtgg acagcaaatg ggtcgggatc tgtacgacga tgacgataag gatctcgcca
     1201 ccatggtcga ctcatcacgt cgtaagtgga ataagacagg tcacgcagtc agagctatag
     1261 gtcggctgag ctcactcgag aacgtctata tcaaggccga caagcagaag aacggcatca
     1321 aggcgaactt caagatccgc cacaacatcg aggacggcgg cgtgcagctc gcctaccact
     1381 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     1441 gcgtgcagtc caaactttcg aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     1501 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag ggcggtaccg
     1561 gagggagcat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     1621 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggt gagggcgatg
     1681 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     1741 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
     1801 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacatc caggagcgca
     1861 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     1921 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     1981 tggggcacaa gctggagtac aacctgccgg accaactgac tgaagagcag atcgcagaat
     2041 ttaaagagga attctcccta tttgacaagg acggggatgg gacaataaca accaaggagc
     2101 tggggacggt gatgcggtct ctggggcaga accccacaga agcagagctg caggacatga
     2161 tcaatgaagt agatgccgac ggtgacggca caatcgactt ccctgagttc ctgacaatga
     2221 tggcaagaaa aatgaaatac agggacacgg aagaagaaat tagagaagcg ttcggtgtgt
     2281 ttgataagga tggcaatggc tacatcagtg cagcagagct tcgccacgtg atgacaaacc
     2341 ttggagagaa gttaacagat gaagaggttg atgaaatgat cagggaagca gacatcgatg
     2401 gggatggtca ggtaaactac gaagagtttg tacaaatgat gacagcgaag tgaggcgcgc
     2461 ccctgcaggg aattcgatat caagcttatc gataatcaac ctctggatta caaaatttgt
     2521 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     2581 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     2641 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     2701 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     2761 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     2821 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     2881 tcggggaaat catcgtcctt tccttggctg ctcgcctatg ttgccacctg gattctgcgc
     2941 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     3001 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     3061 tccctttggg ccgcctcccc gcatcgatac cgagcgctgc tcgagagatc tgtgatagcg
     3121 gccatcaagc tggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt
     3181 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     3241 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     3301 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     3361 caatgtatca gcttatcgat accgcatgca cgtgcggacc gagcggccgc aggaacccct
     3421 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     3481 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
     3541 ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     3601 ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3661 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc
     3721 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3781 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3841 ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3901 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct
     3961 atctcgggct attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa
     4021 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
     4081 ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     4141 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     4201 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     4261 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     4321 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     4381 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     4441 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     4501 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     4561 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     4621 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     4681 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     4741 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     4801 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     4861 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     4921 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     4981 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     5041 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     5101 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     5161 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     5221 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     5281 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     5341 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     5401 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     5461 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     5521 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     5581 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     5641 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     5701 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     5761 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     5821 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     5881 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     5941 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     6001 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     6061 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     6121 gccttttgct ggccttttgc tcacatgt
//
