LOCUS       sequence-421890-        6807 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV construct expressing eYFP-tagged chimeric opsin variant C1V1 
            driven by cholinergic neuron-targeting enhancer. Alias: 
            pAAV_S9E27_C1V1_eYFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6807)
  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 6807)
  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_BiCHATe27_C1V1_eYFP
FEATURES             Location/Qualifiers
     source          1..6807
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          1147..1279
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     CDS             join(2409..2411,2412..3125)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2451..2472)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2712..2731)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3059..3080
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3150..3738
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3203..3223)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3621..3632)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3907..3988
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3944..3963)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3998..4017
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   4067..4207
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4067..4196
                     /label=AAV2 ITR
     rep_origin      4282..4737
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4369..4388)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4579..4600
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4754..4773)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4873..4895
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4933..4951)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5019..5123
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5124..5192,5193..5984)
                     /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(5342..5361)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6155..6743
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6644..6663
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(6805..6807,1..127)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg ggcgtcgggc gacctttggt
       61 cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg
      121 ggttcctgcg gccgcacgcg tatcgatcct cgggtaattt gtaaagacaa tttttctttt
      181 ccccctctga acctaggcct taggctggct aggcaagtct ccactacagc cccgcccact
      241 ccatctcggc tgtggctcgc gggaaaggca gccccctaca gttagtctcc ccagccatct
      301 tctccggttc ccctgatgct ctcatggccc cagccatcct agagctggtg cgccatcgct
      361 cacctttagc attcggcctg ctgggggtga gcttttgggt tagaaggaac aaggcatcag
      421 atggggtcgt gtttttttac attccaatca gttttcggag gggaagaaca gacatttttg
      481 tcagctaatt gcatttaatt acagagacaa tcgctgtttc cgaacgttgt tgtgtccact
      541 ccctcatcct tcaatcagtt caagcacatc cggccccagc tgggagcctt ttaattgcat
      601 aatttggatt tcacttctgg aaagaaactt gagaaagact ccgtgagggt ctccattaac
      661 acaatcagct ccccaggcac ccggggctgg ccctaattga cccatctcag ctaatgcctc
      721 tttccaaaga acgcccgcgt catttctttg gcttgtttgt gctattgtat cctgttttag
      781 tgtcttgatc ttaaacatct ttaaagaggg cactcagtgg cttgtttaac tggaagtggt
      841 ccagctggtt tggaaggaca gtgcatcagg tgatgtaagt ccaagggtag gaatttctgg
      901 cctctccttc ccctgcccat tgcaccgctc tccccaacag gaagtagcta cacatgataa
      961 ggtgcactgt ggcttctagg acttatcgat gtttaaactt cctgcagccc gggctgggca
     1021 taaaagtcag ggcagagcca tctattgctt acatttgctt ctagcctgca agtcgaggag
     1081 cgcagccttc cagaagcaga gcgcggcgcc ttaagctgca gaagttggtc gtgaggcact
     1141 gggcacgtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa actgggcttg
     1201 tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac tgacatccac
     1261 tttgcctttc tctccacagc gagctcgctg ggtaccggta ggccaccatg tcgcggaggc
     1321 catggcttct tgccctagcg ctggcagtgg cgctggcggc cggcagcgca ggagcctcga
     1381 ctggcagtga cgcgacggtg ccggtcgcga ctcaggatgg ccccgactac gttttccacc
     1441 gtgcccacga gcgcatgctc ttccaaacct catacactct tgagaacaat ggttctgtta
     1501 tttgcatccc gaacaacggc cagtgcttct gcttggcttg gcttaaatcc aacggaacaa
     1561 atgccgagaa gttggctgcc aacattctgc agtggattac ttttgcgctt tcagcgctct
     1621 gcctgatgtt ctacggctac cagacctgga agtctacttg cggctgggag accatttacg
     1681 tggccacgat cgagatgatc aagttcatca tcgagtattt ccatgagttt gacgaacctg
     1741 cggtgatcta ctcatccaac ggcaacaaga ccgtgtggct tcgttacgcg acctggctgc
     1801 tcacctgccc ggtccttctg atccatctga gcaacctcac aggcctgaag gacgattata
     1861 gcaaaaggac tatgggcctg ttggtttctg atgtgggatg catcgtgtgg ggcgcaacca
     1921 gcgccatgtg tacggggtgg acgaagatcc tgttcttcct catctcattg agctatggta
     1981 tgtataccta ttttcatgct gctaaagttt atatcgaagc attccacaca gttccaaaag
     2041 ggatttgtcg agaactggtc cgagtgatgg cctggacatt ctttgtggct tggggaatgt
     2101 ttccagtcct gtttctgctg ggcacggaag gattcggtca tatcagccct tatggatctg
     2161 ccattgggca ctccatcctc gacctgattg caaagaacat gtggggtgtg ctggggaatt
     2221 acctgcgcgt caaaatccac gagcacatcc tgttgtatgg cgacatcaga aagaagcaga
     2281 aaattacgat cgccggccaa gagatggagg ttgagacact ggtggctgaa gaggaggacg
     2341 cggccgccaa gagcaggatc accagcgagg gcgagtacat ccccctggac cagatcgaca
     2401 tcaacgtggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     2461 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     2521 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     2581 ccaccctcgt gaccaccttc ggctacggcc tgcagtgctt cgcccgctac cccgaccaca
     2641 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     2701 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     2761 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     2821 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     2881 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     2941 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     3001 accactacct gagctaccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     3061 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     3121 agtaagaatt cgatatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa
     3181 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     3241 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     3301 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     3361 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     3421 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     3481 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     3541 ggaaatcatc gtcctttcct tggctgctcg cctatgttgc cacctggatt ctgcgcggga
     3601 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     3661 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     3721 tttgggccgc ctccccgcat cgataccgag cgctgctcga gagatctgtg atagcggcca
     3781 tcaagctggc cgcgactcta gatcataatc agccatacca catttgtaga ggttttactt
     3841 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     3901 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     3961 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     4021 gtatcagctt atcgataccg catgcacgtg cggaccgagc ggccgcagga acccctagtg
     4081 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag
     4141 gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc
     4201 ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     4261 atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     4321 tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt
     4381 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     4441 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg
     4501 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     4561 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct
     4621 cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg
     4681 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat
     4741 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     4801 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     4861 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     4921 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     4981 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     5041 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     5101 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     5161 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     5221 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     5281 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     5341 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     5401 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     5461 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     5521 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     5581 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     5641 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     5701 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     5761 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     5821 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     5881 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     5941 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     6001 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     6061 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     6121 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     6181 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     6241 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     6301 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6361 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6421 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6481 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6541 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6601 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6661 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6721 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     6781 tttgctggcc ttttgctcac atgtcct
//
