LOCUS       sequence-300331-        7067 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 7067)
  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 7067)
  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..7067
                     /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          836..968
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     951..970
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1009..1018
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1015..1941
                     /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(1957..1959,1960..2670)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(1957..1959,1960..2670)
                     /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             2692..2748
                     /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             2761..2781
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(2782..2784,2785..3498)
                     /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(2782..2784,2785..3495)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     misc_feature    3508..4096
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3561..3581)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3979..3990)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4104..4328
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4330..4470
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4330..4459
                     /label=AAV2 ITR
     rep_origin      4545..5000
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4632..4651)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4842..4863
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5017..5036)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5136..5158
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5196..5214)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5282..5386
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5387..5455,5456..6247)
                     /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(5605..5624)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6418..7006
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6907..6926
                     /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 ctgtttgagc
      181 gggcagagca agtgcgaaaa gattaaaaag tgttctcctc atcctttctg ctcatatgac
      241 cagcgctgca gtgtcgcgcg ccaggcgcac gcccgctcgg cctggctccg cgccaggggg
      301 cgcccggctc agcgcacagc tcagcagctc agtcccgccg caccgccaag ctgagcgggc
      361 cacatgccca tggggtccgg gcattcagga ggctcgccca ggaccccgcc gccaggggcc
      421 atctcctgat ctaaagggcc accagcagag actggagcgt tctgccgctg tcaagggcat
      481 acgggaggca gagaggctcc gatctgggca tctcggacaa tgtcctctcg cgtggggaaa
      541 aaatgatgct gaccagcctc caccccgatg tggcccctgg gctatgcact ctgtctctcc
      601 acgcctccag taccctcttg tcttctgtcc tggatgcaaa tttatgccgc gaaaaatctg
      661 aatgttaaga actatcgatg tttaaacttc ctgcagcccg ggctgggcat aaaagtcagg
      721 gcagagccat ctattgctta catttgcttc tagcctgcaa gtcgaggagc gcagccttcc
      781 agaagcagag cgcggcgcct taagctgcag aagttggtcg tgaggcactg ggcacgtaag
      841 tatcaaggtt acaagacagg tttaaggaga ccaatagaaa ctgggcttgt cgagacagag
      901 aagactcttg cgtttctgat aggcacctat tggtcttact gacatccact ttgcctttct
      961 ctccacaggt gtccactccc agttcaatta cagctcttaa gaaagctagc caccatggac
     1021 tatggcggcg ctttgtctgc cgtcggacgc gaacttttgt tcgttactaa tcctgtggtg
     1081 gtgaacgggt ccgtcctggt ccctgaggat caatgttact gtgccggatg gattgaatct
     1141 cgcggcacga acggcgctca gaccgcgtca aatgtcctgc agtggcttgc agcaggattc
     1201 agcattttgc tgctgatgtt ctatgcctac caaacctgga aatctacatg cggctgggag
     1261 gagatctatg tgtgcgccat tgaaatggtt aaggtgattc tcgagttctt ttttgagttt
     1321 aagaatccct ctatgctcta ccttgccaca ggacaccggg tgcagtggct gcgctatgca
     1381 gagtggctgc tcacttgtcc tgtcatcctt atccgcctga gcaacctcac cggcctgagc
     1441 aacgactaca gcaggagaac catgggactc cttgtctcag acatcgggac tatcgtgtgg
     1501 ggggctacca gcgccatggc aaccggctat gttaaagtca tcttcttttg tcttggattg
     1561 tgctatggcg cgaacacatt ttttcacgcc gccaaagcat atatcgaggg ttatcatact
     1621 gtgccaaagg gtcggtgccg ccaggtcgtg accggcatgg catggctgtt tttcgtgagc
     1681 tggggtatgt tcccaattct cttcattttg gggcccgaag gttttggcgt cctgagcgtc
     1741 tatggctcca ccgtaggtca cacgattatt gatctgatga gtaaaaattg ttgggggttg
     1801 ttgggacact acctgcgcgt cctgatccac gagcacatat tgattcacgg agatatccgc
     1861 aaaaccacca aactgaacat cggcggaacg gagatcgagg tcgagactct cgtcgaagac
     1921 gaagccgagg ccggagccgt gccagctgca gccacagtga gcaagggcga ggagctgttc
     1981 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     2041 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     2101 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     2161 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     2221 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     2281 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     2341 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     2401 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     2461 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     2521 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     2581 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     2641 atcactctcg gcatggacga gctgtacaag tccggagata acggaagcgg agccactaac
     2701 ttctccctgt tgaaacaagc aggggatgtc gaagagaatc ccgggccatc tagaatggcc
     2761 cccaagaaga agaggaaggt ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     2821 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
     2881 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     2941 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
     3001 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     3061 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     3121 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     3181 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
     3241 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
     3301 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     3361 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag
     3421 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     3481 gacgagctgt acaagtaatg gcgcgccaat caacctctgg attacaaaat ttgtgaaaga
     3541 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     3601 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     3661 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     3721 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     3781 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     3841 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     3901 aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     3961 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     4021 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     4081 tgggccgcct ccccgcctgg atcctgtgcc ttctagttgc cagccatctg ttgtttgccc
     4141 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     4201 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     4261 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     4321 ctctatggca ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
     4381 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag
     4441 tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc
     4501 atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg
     4561 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4621 cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4681 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     4741 cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4801 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4861 tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat
     4921 ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     4981 aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata
     5041 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     5101 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     5161 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     5221 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     5281 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     5341 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     5401 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     5461 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     5521 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     5581 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     5641 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     5701 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     5761 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     5821 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     5881 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     5941 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     6001 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     6061 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     6121 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     6181 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     6241 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     6301 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     6361 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     6421 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     6481 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     6541 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     6601 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     6661 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     6721 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     6781 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     6841 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     6901 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     6961 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     7021 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgt
//
