LOCUS       sequence-300338-        7191 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 E29 regulatory 
            element.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7191)
  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 7191)
  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-E29-ChR2GFP2x
FEATURES             Location/Qualifiers
     source          1..7191
                     /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          960..1092
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1075..1094
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1133..1142
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1139..2065
                     /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(2081..2083,2084..2794)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(2081..2083,2084..2794)
                     /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             2816..2872
                     /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             2885..2905
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(2906..2908,2909..3622)
                     /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(2906..2908,2909..3619)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     misc_feature    3632..4220
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3685..3705)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4103..4114)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4228..4452
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4454..4594
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4454..4583
                     /label=AAV2 ITR
     rep_origin      4669..5124
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4756..4775)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4966..4987
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5141..5160)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5260..5282
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5320..5338)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5406..5510
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5511..5579,5580..6371)
                     /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(5729..5748)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6542..7130
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7031..7050
                     /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 cgtatcgata tagtggcgct
      181 atgtgggaga tgtctgggtg gcagagagaa gcatcaggga agatgatggc aagggagcac
      241 tccttctcgg agggagcact ccttctcgga gggtcctgtc ataggactgg ctcctcgctc
      301 ccctttctgg gacagatgga tattaccttc attcctatgc tgccactcag gaaatgggct
      361 ggaggaggag tgacttttcc aaggtcacac agctggggct gagctcagac atccctcctc
      421 cctctcctct gctccccctc ctcccaggca cccgatgtca gctgctctga agtgagtggc
      481 tatttttatt ctggtgcgag ggacctgaaa cagcagagcc atgtaggtta gaagactcta
      541 ggacaccggc ctcagcaggc aaggaaaagc tcactgcagg cacaggaccc tgctaagctt
      601 gcctggggga actcttgttc cttaaagact accctgtcct gggcttctgg gtctccaaac
      661 agttcttcat tagaggtcag aatagagcac agaaccagaa taagccgtgt tcatagccct
      721 catcctgtct ccagctcgaa cactaagctt gaaattgact aagacttgat cccatgccaa
      781 ggcaaagcct ctactctatc gatgtttaaa cttcctgcag cccgggctgg gcataaaagt
      841 cagggcagag ccatctattg cttacatttg cttctagcct gcaagtcgag gagcgcagcc
      901 ttccagaagc agagcgcggc gccttaagct gcagaagttg gtcgtgaggc actgggcacg
      961 taagtatcaa ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac
     1021 agagaagact cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct
     1081 ttctctccac aggtgtccac tcccagttca attacagctc ttaagaaagc tagccaccat
     1141 ggactatggc ggcgctttgt ctgccgtcgg acgcgaactt ttgttcgtta ctaatcctgt
     1201 ggtggtgaac gggtccgtcc tggtccctga ggatcaatgt tactgtgccg gatggattga
     1261 atctcgcggc acgaacggcg ctcagaccgc gtcaaatgtc ctgcagtggc ttgcagcagg
     1321 attcagcatt ttgctgctga tgttctatgc ctaccaaacc tggaaatcta catgcggctg
     1381 ggaggagatc tatgtgtgcg ccattgaaat ggttaaggtg attctcgagt tcttttttga
     1441 gtttaagaat ccctctatgc tctaccttgc cacaggacac cgggtgcagt ggctgcgcta
     1501 tgcagagtgg ctgctcactt gtcctgtcat ccttatccgc ctgagcaacc tcaccggcct
     1561 gagcaacgac tacagcagga gaaccatggg actccttgtc tcagacatcg ggactatcgt
     1621 gtggggggct accagcgcca tggcaaccgg ctatgttaaa gtcatcttct tttgtcttgg
     1681 attgtgctat ggcgcgaaca cattttttca cgccgccaaa gcatatatcg agggttatca
     1741 tactgtgcca aagggtcggt gccgccaggt cgtgaccggc atggcatggc tgtttttcgt
     1801 gagctggggt atgttcccaa ttctcttcat tttggggccc gaaggttttg gcgtcctgag
     1861 cgtctatggc tccaccgtag gtcacacgat tattgatctg atgagtaaaa attgttgggg
     1921 gttgttggga cactacctgc gcgtcctgat ccacgagcac atattgattc acggagatat
     1981 ccgcaaaacc accaaactga acatcggcgg aacggagatc gaggtcgaga ctctcgtcga
     2041 agacgaagcc gaggccggag ccgtgccagc tgcagccaca gtgagcaagg gcgaggagct
     2101 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     2161 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     2221 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
     2281 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     2341 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     2401 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     2461 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     2521 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     2581 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     2641 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct
     2701 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     2761 cgggatcact ctcggcatgg acgagctgta caagtccgga gataacggaa gcggagccac
     2821 taacttctcc ctgttgaaac aagcagggga tgtcgaagag aatcccgggc catctagaat
     2881 ggcccccaag aagaagagga aggtggtgag caagggcgag gagctgttca ccggggtggt
     2941 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     3001 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     3061 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     3121 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     3181 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     3241 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     3301 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     3361 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     3421 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     3481 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     3541 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     3601 catggacgag ctgtacaagt aatggcgcgc caatcaacct ctggattaca aaatttgtga
     3661 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     3721 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     3781 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     3841 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     3901 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     3961 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     4021 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     4081 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     4141 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     4201 cctttgggcc gcctccccgc ctggatcctg tgccttctag ttgccagcca tctgttgttt
     4261 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     4321 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     4381 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     4441 tgggctctat ggcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     4501 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     4561 tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt
     4621 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
     4681 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     4741 gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     4801 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     4861 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     4921 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     4981 aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc
     5041 cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     5101 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
     5161 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     5221 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     5281 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     5341 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     5401 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     5461 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     5521 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     5581 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     5641 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     5701 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     5761 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     5821 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     5881 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     5941 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     6001 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     6061 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     6121 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     6181 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     6241 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     6301 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     6361 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     6421 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     6481 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     6541 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     6601 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     6661 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     6721 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     6781 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     6841 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     6901 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     6961 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     7021 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     7081 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     7141 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg t
//
