LOCUS       sequence-300337-        7179 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 7179)
  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 7179)
  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..7179
                     /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          948..1080
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1063..1082
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1121..1130
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1127..2053
                     /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(2069..2071,2072..2782)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(2069..2071,2072..2782)
                     /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             2804..2860
                     /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             2873..2893
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(2894..2896,2897..3610)
                     /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(2894..2896,2897..3607)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     misc_feature    3620..4208
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3673..3693)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4091..4102)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4216..4440
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4442..4582
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4442..4571
                     /label=AAV2 ITR
     rep_origin      4657..5112
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4744..4763)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4954..4975
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5129..5148)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5248..5270
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5308..5326)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5394..5498
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5499..5567,5568..6359)
                     /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(5717..5736)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6530..7118
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7019..7038
                     /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 cgtatagtgg cgctatgtgg
      181 gagatgtctg ggtggcagag agaagcatca gggaagatga tggcaaggga gcactccttc
      241 tcggagggag cactccttct cggagggtcc tgtcatagga ctggctcctc gctccccttt
      301 ctgggacaga tggatattac cttcattcct atgctgccac tcaggaaatg ggctggagga
      361 ggagtgactt ttccaaggtc acacagctgg ggctgagctc agacatccct cctccctctc
      421 ctctgctccc cctcctccca ggcacccgat gtcagctgct ctgaagtgag tggctatttt
      481 tattctggtg cgagggacct gaaacagcag agccatgtag gttagaagac tctaggacac
      541 cggcctcagc aggcaaggaa aagctcactg caggcacagg accctgctaa gcttgcctgg
      601 gggaactctt gttccttaaa gactaccctg tcctgggctt ctgggtctcc aaacagttct
      661 tcattagagg tcagaataga gcacagaacc agaataagcc gtgttcatag ccctcatcct
      721 gtctccagct cgaacactaa gcttgaaatt gactaagact tgatcccatg ccaaggcaaa
      781 gcctctactc tgtttaaact tcctgcagcc cgggctgggc ataaaagtca gggcagagcc
      841 atctattgct tacatttgct tctagcctgc aagtcgagga gcgcagcctt ccagaagcag
      901 agcgcggcgc cttaagctgc agaagttggt cgtgaggcac tgggcacgta agtatcaagg
      961 ttacaagaca ggtttaagga gaccaataga aactgggctt gtcgagacag agaagactct
     1021 tgcgtttctg ataggcacct attggtctta ctgacatcca ctttgccttt ctctccacag
     1081 gtgtccactc ccagttcaat tacagctctt aagaaagcta gccaccatgg actatggcgg
     1141 cgctttgtct gccgtcggac gcgaactttt gttcgttact aatcctgtgg tggtgaacgg
     1201 gtccgtcctg gtccctgagg atcaatgtta ctgtgccgga tggattgaat ctcgcggcac
     1261 gaacggcgct cagaccgcgt caaatgtcct gcagtggctt gcagcaggat tcagcatttt
     1321 gctgctgatg ttctatgcct accaaacctg gaaatctaca tgcggctggg aggagatcta
     1381 tgtgtgcgcc attgaaatgg ttaaggtgat tctcgagttc ttttttgagt ttaagaatcc
     1441 ctctatgctc taccttgcca caggacaccg ggtgcagtgg ctgcgctatg cagagtggct
     1501 gctcacttgt cctgtcatcc ttatccgcct gagcaacctc accggcctga gcaacgacta
     1561 cagcaggaga accatgggac tccttgtctc agacatcggg actatcgtgt ggggggctac
     1621 cagcgccatg gcaaccggct atgttaaagt catcttcttt tgtcttggat tgtgctatgg
     1681 cgcgaacaca ttttttcacg ccgccaaagc atatatcgag ggttatcata ctgtgccaaa
     1741 gggtcggtgc cgccaggtcg tgaccggcat ggcatggctg tttttcgtga gctggggtat
     1801 gttcccaatt ctcttcattt tggggcccga aggttttggc gtcctgagcg tctatggctc
     1861 caccgtaggt cacacgatta ttgatctgat gagtaaaaat tgttgggggt tgttgggaca
     1921 ctacctgcgc gtcctgatcc acgagcacat attgattcac ggagatatcc gcaaaaccac
     1981 caaactgaac atcggcggaa cggagatcga ggtcgagact ctcgtcgaag acgaagccga
     2041 ggccggagcc gtgccagctg cagccacagt gagcaagggc gaggagctgt tcaccggggt
     2101 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     2161 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
     2221 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt
     2281 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     2341 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     2401 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     2461 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
     2521 tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat
     2581 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     2641 ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc
     2701 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     2761 cggcatggac gagctgtaca agtccggaga taacggaagc ggagccacta acttctccct
     2821 gttgaaacaa gcaggggatg tcgaagagaa tcccgggcca tctagaatgg cccccaagaa
     2881 gaagaggaag gtggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     2941 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     3001 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     3061 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
     3121 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     3181 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     3241 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     3301 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     3361 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     3421 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     3481 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
     3541 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     3601 gtacaagtaa tggcgcgcca atcaacctct ggattacaaa atttgtgaaa gattgactgg
     3661 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     3721 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     3781 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     3841 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     3901 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     3961 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac
     4021 gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     4081 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     4141 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     4201 ctccccgcct ggatcctgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     4261 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     4321 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     4381 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     4441 caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     4501 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag
     4561 cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc
     4621 ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa
     4681 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     4741 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     4801 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     4861 aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     4921 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     4981 cactcaaccc tatctcgggc tattcttttg atttataagg gattttgccg atttcggcct
     5041 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     5101 cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     5161 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     5221 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     5281 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     5341 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     5401 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     5461 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     5521 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     5581 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     5641 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     5701 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     5761 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     5821 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     5881 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     5941 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     6001 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     6061 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     6121 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     6181 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     6241 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     6301 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     6361 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     6421 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     6481 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     6541 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     6601 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     6661 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     6721 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     6781 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     6841 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     6901 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     6961 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     7021 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     7081 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     7141 tacggttcct ggccttttgc tggccttttg ctcacatgt
//
