LOCUS       sequence-300263-        7055 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector to drive the expression of dChr2-GFP-P2A-GFP in PV 
            cortical interneuronsunder the control of the E11 regulatory 
            element.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7055)
  AUTHORS   Vormstein-Schneider D, Lin JD, Pelkey KA, Chittajallu R, Guo B, 
            Arias-Garcia MA, Allaway K, Sakopoulos S, Schneider G, Stevenson O, 
            Vergara J, Sharma J, Zhang Q, Franken TP, Smith J, Ibrahim LA, M 
            Astro KJ, Sabri E, Huang S, Favuzzi E, Burbridge T, Xu Q, Guo L, 
            Vogel I, Sanchez V, Saldi GA, Gorissen BL, Yuan X, Zaghloul KA, 
            Devinsky O, Sabatini BL, Batista-Brito R, Reynolds J, Feng G, Fu Z, 
            McBain CJ, Fishell G, Dimidschstein J
  TITLE     Viral manipulation of functionally distinct interneurons in mice, 
            non-human primates and humans.
  JOURNAL   Nat Neurosci. 2020 Dec;23(12):1629-1636. doi: 
            10.1038/s41593-020-0692-9. Epub 2020 Aug 17.
  PUBMED    32807948
REFERENCE   2  (bases 1 to 7055)
  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: 
            "10.1038/s41593-020-0692-9"; journalName: "Nat Neurosci"; date: 
            "2020-12"; volume: "23"; issue: "12"; pages: "1629-1636"
COMMENT     Sequence Label: pAAV-E11-ChR2GFP2x
FEATURES             Location/Qualifiers
     source          1..7055
                     /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          824..956
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     939..958
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      997..1006
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1003..1929
                     /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             join(1945..1947,1948..2658)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(1945..1947,1948..2658)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             2680..2736
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2749..2769
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(2770..2772,2773..3486)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(2770..2772,2773..3483)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     misc_feature    3496..4084
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3549..3569)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3967..3978)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4092..4316
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4318..4458
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4318..4447
                     /label=AAV2 ITR
     rep_origin      4533..4988
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4620..4639)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4830..4851
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5005..5024)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5124..5146
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5184..5202)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5270..5374
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5375..5443,5444..6235)
                     /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(5593..5612)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6406..6994
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6895..6914
                     /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 gctagactgc agagggcccg cggccgcacg cgtactgttt gagcgggcag
      181 agcaagtgcg aaaagattaa aaagtgttct cctcatcctt tctgctcata tgaccagcgc
      241 tgcagtgtcg cgcgccaggc gcacgcccgc tcggcctggc tccgcgccag ggggcgcccg
      301 gctcagcgca cagctcagca gctcagtccc gccgcaccgc caagctgagc gggccacatg
      361 cccatggggt ccgggcattc aggaggctcg cccaggaccc cgccgccagg ggccatctcc
      421 tgatctaaag ggccaccagc agagactgga gcgttctgcc gctgtcaagg gcatacggga
      481 ggcagagagg ctccgatctg ggcatctcgg acaatgtcct ctcgcgtggg gaaaaaatga
      541 tgctgaccag cctccacccc gatgtggccc ctgggctatg cactctgtct ctccacgcct
      601 ccagtaccct cttgtcttct gtcctggatg caaatttatg ccgcgaaaaa tctgaatgtt
      661 aagaactgtt taaacttcct gcagcccggg ctgggcataa aagtcagggc agagccatct
      721 attgcttaca tttgcttcta gcctgcaagt cgaggagcgc agccttccag aagcagagcg
      781 cggcgcctta agctgcagaa gttggtcgtg aggcactggg cacgtaagta tcaaggttac
      841 aagacaggtt taaggagacc aatagaaact gggcttgtcg agacagagaa gactcttgcg
      901 tttctgatag gcacctattg gtcttactga catccacttt gcctttctct ccacaggtgt
      961 ccactcccag ttcaattaca gctcttaaga aagctagcca ccatggacta tggcggcgct
     1021 ttgtctgccg tcggacgcga acttttgttc gttactaatc ctgtggtggt gaacgggtcc
     1081 gtcctggtcc ctgaggatca atgttactgt gccggatgga ttgaatctcg cggcacgaac
     1141 ggcgctcaga ccgcgtcaaa tgtcctgcag tggcttgcag caggattcag cattttgctg
     1201 ctgatgttct atgcctacca aacctggaaa tctacatgcg gctgggagga gatctatgtg
     1261 tgcgccattg aaatggttaa ggtgattctc gagttctttt ttgagtttaa gaatccctct
     1321 atgctctacc ttgccacagg acaccgggtg cagtggctgc gctatgcaga gtggctgctc
     1381 acttgtcctg tcatccttat ccgcctgagc aacctcaccg gcctgagcaa cgactacagc
     1441 aggagaacca tgggactcct tgtctcagac atcgggacta tcgtgtgggg ggctaccagc
     1501 gccatggcaa ccggctatgt taaagtcatc ttcttttgtc ttggattgtg ctatggcgcg
     1561 aacacatttt ttcacgccgc caaagcatat atcgagggtt atcatactgt gccaaagggt
     1621 cggtgccgcc aggtcgtgac cggcatggca tggctgtttt tcgtgagctg gggtatgttc
     1681 ccaattctct tcattttggg gcccgaaggt tttggcgtcc tgagcgtcta tggctccacc
     1741 gtaggtcaca cgattattga tctgatgagt aaaaattgtt gggggttgtt gggacactac
     1801 ctgcgcgtcc tgatccacga gcacatattg attcacggag atatccgcaa aaccaccaaa
     1861 ctgaacatcg gcggaacgga gatcgaggtc gagactctcg tcgaagacga agccgaggcc
     1921 ggagccgtgc cagctgcagc cacagtgagc aagggcgagg agctgttcac cggggtggtg
     1981 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     2041 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     2101 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     2161 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     2221 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     2281 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     2341 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     2401 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     2461 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     2521 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     2581 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     2641 atggacgagc tgtacaagtc cggagataac ggaagcggag ccactaactt ctccctgttg
     2701 aaacaagcag gggatgtcga agagaatccc gggccatcta gaatggcccc caagaagaag
     2761 aggaaggtgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     2821 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     2881 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     2941 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
     3001 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     3061 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     3121 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     3181 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag
     3241 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
     3301 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     3361 aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
     3421 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     3481 aagtaatggc gcgccaatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     3541 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     3601 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     3661 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     3721 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     3781 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     3841 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc
     3901 tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     3961 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     4021 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     4081 ccgcctggat cctgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     4141 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     4201 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     4261 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcagg
     4321 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
     4381 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag
     4441 cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     4501 tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc
     4561 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     4621 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     4681 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     4741 acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     4801 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     4861 caaccctatc tcgggctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     4921 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt
     4981 tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     5041 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     5101 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     5161 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     5221 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     5281 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     5341 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     5401 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     5461 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     5521 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     5581 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     5641 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     5701 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     5761 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     5821 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     5881 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     5941 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     6001 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     6061 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     6121 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     6181 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     6241 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     6301 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     6361 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     6421 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     6481 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     6541 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     6601 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     6661 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     6721 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     6781 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     6841 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     6901 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     6961 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     7021 gttcctggcc ttttgctggc cttttgctca catgt
//
