LOCUS       sequence-421865-        6448 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV construct expressing mCherry-tagged channelrhodopsin-2 driven by
            Lamp5 interneuron-targeting enhancer. Alias: 
            pAAV_WDL0003_ChR2_mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6448)
  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 6448)
  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_ChR2_mCherry
FEATURES             Location/Qualifiers
     source          1..6448
                     /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      1096..1105
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1102..2028
                     /codon_start=1
                     /product="H134R mutant of an N-terminal fragment of 
                     channelrhodopsin-2 from the alga Chlamydomonas reinhardtii 
                     (Nagel et al., 2005)"
                     /label=ChR2(H134R)
                     /note="blue light-activated cation channel"
                     /translation="MDYGGALSAVGRELLFVTNPVVVNGSVLVPEDQCYCAGWIESRGT
                     NGAQTASNVLQWLAAGFSILLLMFYAYQTWKSTCGWEEIYVCAIEMVKVILEFFFEFKN
                     PSMLYLATGHRVQWLRYAEWLLTCPVILIRLSNLTGLSNDYSRRTMGLLVSDIGTIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     CDS             2041..2748
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(2183..2201)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2451..2470
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2794..3382
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2847..2867)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3265..3276)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3551..3632
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3588..3607)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3642..3661
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   3711..3851
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3711..3840
                     /label=AAV2 ITR
     rep_origin      3926..4381
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4013..4032)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4223..4244
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4398..4417)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4517..4539
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4577..4595)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4663..4767
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4768..4836,4837..5628)
                     /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(4986..5005)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5799..6387
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6288..6307
                     /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 atcgcgccac catggactat ggcggcgctt tgtctgccgt cggacgcgaa
     1141 cttttgttcg ttactaatcc tgtggtggtg aacgggtccg tcctggtccc tgaggatcaa
     1201 tgttactgtg ccggatggat tgaatctcgc ggcacgaacg gcgctcagac cgcgtcaaat
     1261 gtcctgcagt ggcttgcagc aggattcagc attttgctgc tgatgttcta tgcctaccaa
     1321 acctggaaat ctacatgcgg ctgggaggag atctatgtgt gcgccattga aatggttaag
     1381 gtgattctcg agttcttttt tgagtttaag aatccctcta tgctctacct tgccacagga
     1441 caccgggtgc agtggctgcg ctatgcagag tggctgctca cttgtcctgt catccttatc
     1501 cgcctgagca acctcaccgg cctgagcaac gactacagca ggagaaccat gggactcctt
     1561 gtctcagaca tcgggactat cgtgtggggg gctaccagcg ccatggcaac cggctatgtt
     1621 aaagtcatct tcttttgtct tggattgtgc tatggcgcga acacattttt tcacgccgcc
     1681 aaagcatata tcgagggtta tcatactgtg ccaaagggtc ggtgccgcca ggtcgtgacc
     1741 ggcatggcat ggctgttttt cgtgagctgg ggtatgttcc caattctctt cattttgggg
     1801 cccgaaggtt ttggcgtcct gagcgtctat ggctccaccg taggtcacac gattattgat
     1861 ctgatgagta aaaattgttg ggggttgttg ggacactacc tgcgcgtcct gatccacgag
     1921 cacatattga ttcacggaga tatccgcaaa accaccaaac tgaacatcgg cggaacggag
     1981 atcgaggtcg agactctcgt cgaagacgaa gccgaggccg gagccgtgcc agcggccgcc
     2041 gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtg
     2101 cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc
     2161 ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc
     2221 gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt gaagcacccc
     2281 gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg
     2341 atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc
     2401 gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg
     2461 cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc
     2521 ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta cgacgctgag
     2581 gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta caacgtcaac
     2641 atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc
     2701 gccgagggcc gccactccac cggcggcatg gacgagctgt acaagtaagg cgcgccccac
     2761 ccctgcaggg aattcgatat caagcttatc gataatcaac ctctggatta caaaatttgt
     2821 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     2881 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     2941 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     3001 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     3061 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     3121 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     3181 tcggggaaat catcgtcctt tccttggctg ctcgcctatg ttgccacctg gattctgcgc
     3241 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     3301 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     3361 tccctttggg ccgcctcccc gcatcgatac cgagcgctgc tcgagagatc tgtgatagcg
     3421 gccatcaagc tggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt
     3481 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     3541 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     3601 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     3661 caatgtatca gcttatcgat accgcatgca cgtgcggacc gagcggccgc aggaacccct
     3721 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     3781 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
     3841 ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     3901 ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3961 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc
     4021 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     4081 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     4141 ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     4201 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct
     4261 atctcgggct attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa
     4321 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
     4381 ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     4441 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     4501 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     4561 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     4621 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     4681 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     4741 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     4801 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     4861 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     4921 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     4981 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     5041 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     5101 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     5161 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     5221 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     5281 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     5341 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     5401 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     5461 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     5521 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     5581 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     5641 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     5701 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     5761 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     5821 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     5881 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     5941 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     6001 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     6061 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     6121 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     6181 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     6241 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     6301 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     6361 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     6421 gccttttgct ggccttttgc tcacatgt
//
