LOCUS       sequence-425143-        5740 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ChR2(L132C, H134R, T159C)-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 5740)
  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 5740)
  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: AiP13278:
            pAAV-AiE0779m_3xC2-minBG-ChR2(CRC)-E...
FEATURES             Location/Qualifiers
     source          1..5740
                     /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      816..825
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             822..1748
                     /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
                     PSMLYLATGHRVQWLRYAEWLLTCPVICIRLSNLTGLSNDYSRRTMGLLVSDIGCIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     regulatory      1758..1767
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1764..1766,1767..1769,1770..2483)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1809..1830)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2070..2089)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2417..2438
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2567..2587)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2774..2885
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3003..3143
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3003..3132
                     /label=AAV2 ITR
     rep_origin      3218..3673
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3305..3324)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3515..3536
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3690..3709)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3809..3831
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3869..3887)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3955..4059
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4060..4128,4129..4920)
                     /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(4278..4297)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5091..5679
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5580..5599
                     /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 gaagctagcg cggccgccac catggactat ggcggcgctt
      841 tgtctgccgt cggacgcgaa cttttgttcg ttactaatcc tgtggtggtg aacgggtccg
      901 tcctggtccc tgaggatcaa tgttactgtg ccggatggat tgaatctcgc ggcacgaacg
      961 gcgctcagac cgcgtcaaat gtcctgcagt ggcttgcagc aggattcagc attttgctgc
     1021 tgatgttcta tgcctaccaa acctggaaat ctacatgcgg ctgggaggag atctatgtgt
     1081 gcgccattga aatggttaag gtgattctcg agttcttttt tgagtttaag aatccctcta
     1141 tgctctacct tgccacagga caccgggtgc agtggctgcg ctatgcagag tggctgctca
     1201 cttgtcctgt catctgcatc cgcctgagca acctcaccgg cctgagcaac gactacagca
     1261 ggagaaccat gggactcctt gtctcagaca tcgggtgtat cgtgtggggg gctaccagcg
     1321 ccatggcaac cggctatgtt aaagtcatct tcttttgtct tggattgtgc tatggcgcga
     1381 acacattttt tcacgccgcc aaagcatata tcgagggtta tcatactgtg ccaaagggtc
     1441 ggtgccgcca ggtcgtgacc ggcatggcat ggctgttttt cgtgagctgg ggtatgttcc
     1501 caattctctt cattttgggg cccgaaggtt ttggcgtcct gagcgtctat ggctccaccg
     1561 taggtcacac gattattgat ctgatgagta aaaattgttg ggggttgttg ggacactacc
     1621 tgcgcgtcct gatccacgag cacatattga ttcacggaga tatccgcaaa accaccaaac
     1681 tgaacatcgg cggaacggag atcgaggtcg agactctcgt cgaagacgaa gccgaggccg
     1741 gagccgtgcc agcggccgcc accatggtga gcaagggcga ggagctgttc accggggtgg
     1801 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     1861 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     1921 agctgcccgt gccctggccc accctcgtga ccaccttcgg ctacggcctg cagtgcttcg
     1981 cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     2041 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     2101 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     2161 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     2221 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
     2281 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     2341 ccgtgctgct gcccgacaac cactacctga gctaccagtc cgccctgagc aaagacccca
     2401 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     2461 gcatggacga gctgtacaag taaggcgcgc cgcggccgcg aattcgatat cataatcaac
     2521 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     2581 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     2641 tcattttctc ctccttgtat aaatcctggt tagttcttgc cacggcggaa ctcatcgccg
     2701 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggctc
     2761 gagagatctt cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     2821 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     2881 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     2941 aagggggagg attgggaaga caatagcagg catgagatct cacgtgcgga ccgagcggcc
     3001 gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
     3061 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga
     3121 gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg
     3181 cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta
     3241 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg
     3301 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     3361 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     3421 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     3481 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     3541 acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc
     3601 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     3661 acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     3721 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     3781 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     3841 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     3901 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     3961 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4021 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     4081 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     4141 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     4201 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     4261 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     4321 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     4381 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     4441 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     4501 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     4561 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     4621 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     4681 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     4741 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     4801 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     4861 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     4921 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     4981 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5041 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5101 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5161 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5221 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     5281 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5341 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     5401 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     5461 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     5521 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     5581 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     5641 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     5701 ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
