LOCUS       sequence-421843-        6218 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV construct expressing mCherry-tagged channelrhodopsin-2 driven by
            VIP interneuron-targeting enhancer. Alias: pAAV_YWE4.2_ChR2_mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6218)
  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 6218)
  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_ChR2_mCherry
FEATURES             Location/Qualifiers
     source          1..6218
                     /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      866..875
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             872..1798
                     /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             1811..2518
                     /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(1953..1971)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2221..2240
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2564..3152
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2617..2637)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3035..3046)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3321..3402
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3358..3377)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3412..3431
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   3481..3621
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3481..3610
                     /label=AAV2 ITR
     rep_origin      3696..4151
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3783..3802)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3993..4014
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4168..4187)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4287..4309
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4347..4365)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4433..4537
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4538..4606,4607..5398)
                     /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(4756..4775)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5569..6157
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6058..6077
                     /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 atcgcgccac catggactat ggcggcgctt tgtctgccgt
      901 cggacgcgaa cttttgttcg ttactaatcc tgtggtggtg aacgggtccg tcctggtccc
      961 tgaggatcaa tgttactgtg ccggatggat tgaatctcgc ggcacgaacg gcgctcagac
     1021 cgcgtcaaat gtcctgcagt ggcttgcagc aggattcagc attttgctgc tgatgttcta
     1081 tgcctaccaa acctggaaat ctacatgcgg ctgggaggag atctatgtgt gcgccattga
     1141 aatggttaag gtgattctcg agttcttttt tgagtttaag aatccctcta tgctctacct
     1201 tgccacagga caccgggtgc agtggctgcg ctatgcagag tggctgctca cttgtcctgt
     1261 catccttatc cgcctgagca acctcaccgg cctgagcaac gactacagca ggagaaccat
     1321 gggactcctt gtctcagaca tcgggactat cgtgtggggg gctaccagcg ccatggcaac
     1381 cggctatgtt aaagtcatct tcttttgtct tggattgtgc tatggcgcga acacattttt
     1441 tcacgccgcc aaagcatata tcgagggtta tcatactgtg ccaaagggtc ggtgccgcca
     1501 ggtcgtgacc ggcatggcat ggctgttttt cgtgagctgg ggtatgttcc caattctctt
     1561 cattttgggg cccgaaggtt ttggcgtcct gagcgtctat ggctccaccg taggtcacac
     1621 gattattgat ctgatgagta aaaattgttg ggggttgttg ggacactacc tgcgcgtcct
     1681 gatccacgag cacatattga ttcacggaga tatccgcaaa accaccaaac tgaacatcgg
     1741 cggaacggag atcgaggtcg agactctcgt cgaagacgaa gccgaggccg gagccgtgcc
     1801 agcggccgcc gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg
     1861 cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg
     1921 cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc
     1981 cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt
     2041 gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg
     2101 ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct
     2161 gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg
     2221 ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga
     2281 ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta
     2341 cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta
     2401 caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca
     2461 gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt acaagtaagg
     2521 cgcgccccac ccctgcaggg aattcgatat caagcttatc gataatcaac ctctggatta
     2581 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     2641 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     2701 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     2761 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     2821 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     2881 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     2941 cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctatg ttgccacctg
     3001 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     3061 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     3121 gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgagcgctgc tcgagagatc
     3181 tgtgatagcg gccatcaagc tggccgcgac tctagatcat aatcagccat accacatttg
     3241 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     3301 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     3361 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     3421 ccaaactcat caatgtatca gcttatcgat accgcatgca cgtgcggacc gagcggccgc
     3481 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     3541 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
     3601 gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg
     3661 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag
     3721 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc
     3781 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     3841 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     3901 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     3961 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     4021 actcaactct atctcgggct attcttttga tttataaggg attttgccga tttcggtcta
     4081 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     4141 gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     4201 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     4261 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     4321 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     4381 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     4441 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     4501 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     4561 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     4621 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     4681 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     4741 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     4801 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     4861 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     4921 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     4981 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     5041 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     5101 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     5161 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     5221 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     5281 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     5341 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     5401 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5461 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5521 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5581 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5641 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     5701 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5761 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     5821 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5881 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5941 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6001 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6061 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6121 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     6181 acggttcctg gccttttgct ggccttttgc tcacatgt
//
