LOCUS       sequence-425141-        5735 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ChR2(H134R)-EYFP in striatal direct pathway medium 
            spiny neurons (D1 MSNs).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5735)
  AUTHORS   Hunker AC, Wirthlin ME, Gill G, Johansen NJ, Hooper M, Omstead V, 
            Vargas S, Lerma MN, Taskin N, Weed N, Laird WD, Bishaw YM, Bendrick 
            JL, Gore BB, Ben-Simon Y, Opitz-Araya X, Martinez RA, Way SW, 
            Thyagarajan B, Otto S, Sanchez REA, Alexander JR, Amaya A, Amster A,
            Arbuckle J, Ayala A, Baker PM, Barcelli T, Barta S, Bertagnolli D, 
            Bielstein C, Bishwakarma P, Bowlus J, Boyer G, Brouner K, Casian B, 
            Casper T, Chakka AB, Chakrabarty R, Chong P, Clark M, Colbert K, 
            Daniel S, Dawe T, Departee M, DiValentin P, Donadio NP, Dotson NI, 
            Dwivedi D, Egdorf T, Fliss T, Gary A, Goldy J, Grasso C, Groce EL, 
            Gudsnuk K, Han W, Haradon Z, Hastings S, Helback O, Ho WV, Huang C, 
            Johnson T, Jones DL, Juneau Z, Kenney J, Leibly M, Li S, Liang E, 
            Loeffler H, Lusk NA, Madigan Z, Malloy J, Malone J, McCue R, Melchor
            J, Mich JK, Moosman S, Morin E, Naidoo R, Newman D, Ngo K, Nguyen K,
            Oster AL, Ouellette B, Oyama AA, Pena N, Pham T, Phillips E, Pom C, 
            Potekhina L, Ransford S, Ray PL, Reding M, Rette DF, Reynoldson C, 
            Rimorin C, Sigler AR, Rocha DB, Ronellenfitch K, Ruiz A, Sawyer L, 
            Sevigny JP, Shapovalova NV, Shepard N, Shulga L, Soliman S, Staats 
            B, Taormina MJ, Tieu M, Wang Y, Wilkes J, Wood T, Zhou T, Williford 
            A, Dee N, Mollenkopf T, Ng L, Esposito L, Kalmbach BE, Yao S, Ariza 
            J, Collman F, Mufti S, Smith K, Waters J, Ersing I, Patrick M, Zeng 
            H, Lein ES, Kojima Y, Horwitz G, Owen SF, Levi BP, Daigle TL, Tasic 
            B, Bakken TE, Ting JT
  TITLE     Enhancer AAV toolbox for accessing and perturbing striatal cell 
            types and circuits.
  JOURNAL   Neuron. 2025 May 21;113(10):1507-1524.e17. doi: 
            10.1016/j.neuron.2025.04.035.
  PUBMED    40403704
REFERENCE   2  (bases 1 to 5735)
  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.2025.04"; journalName: "Neuron"; date: 
            "2025-05-21- 21"; volume: "113"; issue: "10"; pages: "1507-1524.e17"
COMMENT     Sequence Label: AiP13276:
            pAAV-AiE0779m_3xC2-minBG-ChR2(H134R)...
FEATURES             Location/Qualifiers
     source          1..5735
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      808..817
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             814..1740
                     /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"
     regulatory      1750..1759
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1756..1758,1759..1761,1762..2475)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1801..1822)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2062..2081)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2409..2430
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2562..2582)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2769..2880
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2998..3138
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2998..3127
                     /label=AAV2 ITR
     rep_origin      3213..3668
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3300..3319)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3510..3531
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3685..3704)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3804..3826
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3864..3882)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3950..4054
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4055..4123,4124..4915)
                     /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(4273..4292)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5086..5674
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5575..5594
                     /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 gcgtaagttt gagacaaatt aaggaaaagc gggttgggag
      181 gagtctgagg ctgtgggtgg ctgcctcgag aggctgctca caacctgaca tgtttcagat
      241 cttttgacac ggtcatgcaa gcaacagaaa cttatatggt ttttagtctc ccaggtagca
      301 gggaaatgat gatgtgggag gattaggggg gggaaaataa gtttgagaca aattaaggaa
      361 aagcgggttg ggaggagtct gaggctgtgg gtggctgcct cgagaggctg ctcacaacct
      421 gacatgtttc agatcttttg acacggtcat gcaagcaaca gaaacttata tggtttttag
      481 tctcccaggt agcagggaaa tgatgatgtg ggaggattag gggggggaaa ataagtttga
      541 gacaaattaa ggaaaagcgg gttgggagga gtctgaggct gtgggtggct gcctcgagag
      601 gctgctcaca acctgacatg tttcagatct tttgacacgg tcatgcaagc aacagaaact
      661 tatatggttt ttagtctccc aggtagcagg gaaatgatga tgtgggagga ttaggggggg
      721 gaaaatgagc tcgggctggg cataaaagtc agggcagagc catctattgc ttacatttgc
      781 ttctgggatc cagatctttc gaagctagcc accatggact atggcggcgc tttgtctgcc
      841 gtcggacgcg aacttttgtt cgttactaat cctgtggtgg tgaacgggtc cgtcctggtc
      901 cctgaggatc aatgttactg tgccggatgg attgaatctc gcggcacgaa cggcgctcag
      961 accgcgtcaa atgtcctgca gtggcttgca gcaggattca gcattttgct gctgatgttc
     1021 tatgcctacc aaacctggaa atctacatgc ggctgggagg agatctatgt gtgcgccatt
     1081 gaaatggtta aggtgattct cgagttcttt tttgagttta agaatccctc tatgctctac
     1141 cttgccacag gacaccgggt gcagtggctg cgctatgcag agtggctgct cacttgtcct
     1201 gtcatcctta tccgcctgag caacctcacc ggcctgagca acgactacag caggagaacc
     1261 atgggactcc ttgtctcaga catcgggact atcgtgtggg gggctaccag cgccatggca
     1321 accggctatg ttaaagtcat cttcttttgt cttggattgt gctatggcgc gaacacattt
     1381 tttcacgccg ccaaagcata tatcgagggt tatcatactg tgccaaaggg tcggtgccgc
     1441 caggtcgtga ccggcatggc atggctgttt ttcgtgagct ggggtatgtt cccaattctc
     1501 ttcattttgg ggcccgaagg ttttggcgtc ctgagcgtct atggctccac cgtaggtcac
     1561 acgattattg atctgatgag taaaaattgt tgggggttgt tgggacacta cctgcgcgtc
     1621 ctgatccacg agcacatatt gattcacgga gatatccgca aaaccaccaa actgaacatc
     1681 ggcggaacgg agatcgaggt cgagactctc gtcgaagacg aagccgaggc cggagccgtg
     1741 ccagcggccg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     1801 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     1861 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     1921 gtgccctggc ccaccctcgt gaccaccttc ggctacggcc tgcagtgctt cgcccgctac
     1981 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     2041 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     2101 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     2161 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     2221 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     2281 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     2341 ctgcccgaca accactacct gagctaccag tccgccctga gcaaagaccc caacgagaag
     2401 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     2461 gagctgtaca agtaataggg cgcgccgcgg ccgcgaattc gatatcataa tcaacctctg
     2521 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     2581 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     2641 ttctcctcct tgtataaatc ctggttagtt cttgccacgg cggaactcat cgccgcctgc
     2701 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggctcgagag
     2761 atcttcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     2821 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     2881 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     2941 ggaggattgg gaagacaata gcaggcatga gatctcacgt gcggaccgag cggccgcagg
     3001 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
     3061 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag
     3121 cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     3181 tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc
     3241 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc
     3301 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     3361 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     3421 acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3481 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3541 caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg
     3601 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt
     3661 tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     3721 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     3781 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     3841 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     3901 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     3961 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     4021 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     4081 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     4141 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     4201 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     4261 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     4321 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     4381 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     4441 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     4501 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     4561 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     4621 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     4681 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     4741 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     4801 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     4861 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     4921 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     4981 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     5041 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     5101 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     5161 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5221 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     5281 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5341 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5401 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5461 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     5521 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     5581 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     5641 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     5701 gttcctggcc ttttgctggc cttttgctca catgt
//
