LOCUS       sequence-421845-        6577 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV construct expressing mCherry-tagged channelrhodopsin-2 driven by
            SST interneuron-targeting enhancer. Alias: pAAV_S9E10_ChR2_mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6577)
  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 6577)
  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_BiSSTe10_ChR2_mCherry
FEATURES             Location/Qualifiers
     source          1..6577
                     /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          1071..1203
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     regulatory      1225..1234
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1231..2157
                     /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             2170..2877
                     /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(2312..2330)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2580..2599
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2923..3511
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2976..2996)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3394..3405)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3680..3761
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3717..3736)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3771..3790
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   3840..3980
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3840..3969
                     /label=AAV2 ITR
     rep_origin      4055..4510
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4142..4161)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4352..4373
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4527..4546)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4646..4668
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4706..4724)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4792..4896
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4897..4965,4966..5757)
                     /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(5115..5134)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5928..6516
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6417..6436
                     /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 ctaggttcct gtctgcaagc atagcagggt
      181 atcattaata gtgtcaggag ttggctctct cccataggat gggtctcaag ttggggcaat
      241 cattggttgg ctgatccctt ggtctctgtc ccatctttat tcctgcacat cttgtaggca
      301 ggacaaattt tggtttgttc caaagtgatt tttaaagcat taccattttg tgttttgttg
      361 gtttaaaaca ctttagctat tctggtgggt gcgtgatcct gcctcactac ggtctcagtc
      421 tccatctcga tggtgactaa ggatgcctcc tatgcactta tttaccatct gaatggttcc
      481 tttatagaat gcatgactat tcaatttttt tgctcattgg acagcatttt aatgtgtgaa
      541 aggattatac atccttcaag atatggaaga aaaagcacaa agccctaaat tctgtgaatt
      601 cagatgtgta aaaaaagaca aggaagtgca aactatatca gcttggtgtc tcattctggg
      661 tggctgtcag agccctgtga gctgacttaa gaattgggtc cctgcaatgg gtgctggcac
      721 ttgcaagggg atcatgagct caaggccata ctgagctgca taatgagacc ttgtctcagt
      781 aactagactg gggttggagg tcagtactaa agggctagcc tagcctaaat tcccgagttc
      841 cacccccagc actataaaag gaaaaggagg aagaggtgga gggaaataaa gaaaaggaaa
      901 gccagtgtat cgatgtttaa acttcctgca gcccgggctg ggcataaaag tcagggcaga
      961 gccatctatt gcttacattt gcttctagcc tgcaagtcga ggagcgcagc cttccagaag
     1021 cagagcgcgg cgccttaagc tgcagaagtt ggtcgtgagg cactgggcac gtaagtatca
     1081 aggttacaag acaggtttaa ggagaccaat agaaactggg cttgtcgaga cagagaagac
     1141 tcttgcgttt ctgataggca cctattggtc ttactgacat ccactttgcc tttctctcca
     1201 cagcgagctc gctagcgcga tcgcgccacc atggactatg gcggcgcttt gtctgccgtc
     1261 ggacgcgaac ttttgttcgt tactaatcct gtggtggtga acgggtccgt cctggtccct
     1321 gaggatcaat gttactgtgc cggatggatt gaatctcgcg gcacgaacgg cgctcagacc
     1381 gcgtcaaatg tcctgcagtg gcttgcagca ggattcagca ttttgctgct gatgttctat
     1441 gcctaccaaa cctggaaatc tacatgcggc tgggaggaga tctatgtgtg cgccattgaa
     1501 atggttaagg tgattctcga gttctttttt gagtttaaga atccctctat gctctacctt
     1561 gccacaggac accgggtgca gtggctgcgc tatgcagagt ggctgctcac ttgtcctgtc
     1621 atccttatcc gcctgagcaa cctcaccggc ctgagcaacg actacagcag gagaaccatg
     1681 ggactccttg tctcagacat cgggactatc gtgtgggggg ctaccagcgc catggcaacc
     1741 ggctatgtta aagtcatctt cttttgtctt ggattgtgct atggcgcgaa cacatttttt
     1801 cacgccgcca aagcatatat cgagggttat catactgtgc caaagggtcg gtgccgccag
     1861 gtcgtgaccg gcatggcatg gctgtttttc gtgagctggg gtatgttccc aattctcttc
     1921 attttggggc ccgaaggttt tggcgtcctg agcgtctatg gctccaccgt aggtcacacg
     1981 attattgatc tgatgagtaa aaattgttgg gggttgttgg gacactacct gcgcgtcctg
     2041 atccacgagc acatattgat tcacggagat atccgcaaaa ccaccaaact gaacatcggc
     2101 ggaacggaga tcgaggtcga gactctcgtc gaagacgaag ccgaggccgg agccgtgcca
     2161 gcggccgccg tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc
     2221 ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
     2281 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc
     2341 ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg
     2401 aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg
     2461 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg
     2521 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc
     2581 cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag
     2641 gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac
     2701 gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac
     2761 aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag
     2821 tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtaaggc
     2881 gcgccccacc cctgcaggga attcgatatc aagcttatcg ataatcaacc tctggattac
     2941 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     3001 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     3061 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     3121 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     3181 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     3241 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     3301 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctatgt tgccacctgg
     3361 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     3421 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     3481 agtcggatct ccctttgggc cgcctccccg catcgatacc gagcgctgct cgagagatct
     3541 gtgatagcgg ccatcaagct ggccgcgact ctagatcata atcagccata ccacatttgt
     3601 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     3661 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     3721 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     3781 caaactcatc aatgtatcag cttatcgata ccgcatgcac gtgcggaccg agcggccgca
     3841 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     3901 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg
     3961 agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     4021 tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc
     4081 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc
     4141 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     4201 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     4261 aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     4321 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     4381 ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat
     4441 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     4501 tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     4561 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     4621 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     4681 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     4741 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     4801 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     4861 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     4921 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     4981 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     5041 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     5101 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     5161 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     5221 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     5281 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     5341 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     5401 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     5461 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     5521 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     5581 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     5641 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     5701 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     5761 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     5821 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5881 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     5941 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     6001 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     6061 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     6121 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     6181 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     6241 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     6301 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     6361 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     6421 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     6481 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     6541 cggttcctgg ccttttgctg gccttttgct cacatgt
//
