LOCUS       sequence-426042-        5750 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ChR2(H134R)-EYFP in striatal medium spiny neurons .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5750)
  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 5750)
  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: AiP14823:
            pAAV-AiE0441h_3xC2-minBG-ChR2(H134R)...
FEATURES             Location/Qualifiers
     source          1..5750
                     /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      823..832
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             829..1755
                     /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      1765..1774
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1771..1773,1774..1776,1777..2490)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1816..1837)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2077..2096)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2424..2445
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2577..2597)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2784..2895
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3013..3153
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3013..3142
                     /label=AAV2 ITR
     rep_origin      3228..3683
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3315..3334)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3525..3546
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3700..3719)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3819..3841
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3879..3897)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3965..4069
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4070..4138,4139..4930)
                     /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(4288..4307)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5101..5689
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5590..5609
                     /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 gcgtcccaca cagcccccac cacctggccg cccacctgaa
      181 aaacttttcc ccaagttctt gagaagtcgc gtaggatcca ggagagaaaa atcaaagtaa
      241 taaaacctca tttaattcct aacggagggc agagctgcga gctgcaatta tcttaatgct
      301 gagctatttt tactctctct ccatctctgt cttctgcctt tctcccacac agcccccacc
      361 acctggccgc ccacctgaaa aacttttccc caagttcttg agaagtcgcg taggatccag
      421 gagagaaaaa tcaaagtaat aaaacctcat ttaattccta acggagggca gagctgcgag
      481 ctgcaattat cttaatgctg agctattttt actctctctc catctctgtc ttctgccttt
      541 ctcccacaca gcccccacca cctggccgcc cacctgaaaa acttttcccc aagttcttga
      601 gaagtcgcgt aggatccagg agagaaaaat caaagtaata aaacctcatt taattcctaa
      661 cggagggcag agctgcgagc tgcaattatc ttaatgctga gctattttta ctctctctcc
      721 atctctgtct tctgcctttc tgagctcggg ctgggcataa aagtcagggc agagccatct
      781 attgcttaca tttgcttctg ggatccagat ctttcgaagc tagccaccat ggactatggc
      841 ggcgctttgt ctgccgtcgg acgcgaactt ttgttcgtta ctaatcctgt ggtggtgaac
      901 gggtccgtcc tggtccctga ggatcaatgt tactgtgccg gatggattga atctcgcggc
      961 acgaacggcg ctcagaccgc gtcaaatgtc ctgcagtggc ttgcagcagg attcagcatt
     1021 ttgctgctga tgttctatgc ctaccaaacc tggaaatcta catgcggctg ggaggagatc
     1081 tatgtgtgcg ccattgaaat ggttaaggtg attctcgagt tcttttttga gtttaagaat
     1141 ccctctatgc tctaccttgc cacaggacac cgggtgcagt ggctgcgcta tgcagagtgg
     1201 ctgctcactt gtcctgtcat ccttatccgc ctgagcaacc tcaccggcct gagcaacgac
     1261 tacagcagga gaaccatggg actccttgtc tcagacatcg ggactatcgt gtggggggct
     1321 accagcgcca tggcaaccgg ctatgttaaa gtcatcttct tttgtcttgg attgtgctat
     1381 ggcgcgaaca cattttttca cgccgccaaa gcatatatcg agggttatca tactgtgcca
     1441 aagggtcggt gccgccaggt cgtgaccggc atggcatggc tgtttttcgt gagctggggt
     1501 atgttcccaa ttctcttcat tttggggccc gaaggttttg gcgtcctgag cgtctatggc
     1561 tccaccgtag gtcacacgat tattgatctg atgagtaaaa attgttgggg gttgttggga
     1621 cactacctgc gcgtcctgat ccacgagcac atattgattc acggagatat ccgcaaaacc
     1681 accaaactga acatcggcgg aacggagatc gaggtcgaga ctctcgtcga agacgaagcc
     1741 gaggccggag ccgtgccagc ggccgccacc atggtgagca agggcgagga gctgttcacc
     1801 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     1861 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     1921 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccttcggcta cggcctgcag
     1981 tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     2041 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     2101 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     2161 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     2221 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     2281 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     2341 gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccgc cctgagcaaa
     2401 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     2461 actctcggca tggacgagct gtacaagtaa tagggcgcgc cgcggccgcg aattcgatat
     2521 cataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     2581 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     2641 cgtatggctt tcattttctc ctccttgtat aaatcctggt tagttcttgc cacggcggaa
     2701 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     2761 tccgtggctc gagagatctt cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     2821 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     2881 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     2941 gcaggacagc aagggggagg attgggaaga caatagcagg catgagatct cacgtgcgga
     3001 ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     3061 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     3121 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta
     3181 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag
     3241 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3301 cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3361 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     3421 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata
     3481 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3541 aactggaaca acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc
     3601 gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     3661 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc
     3721 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     3781 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     3841 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     3901 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     3961 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     4021 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     4081 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     4141 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     4201 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     4261 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     4321 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     4381 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     4441 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     4501 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     4561 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     4621 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     4681 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     4741 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     4801 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     4861 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     4921 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     4981 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     5041 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5101 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5161 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     5221 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     5281 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     5341 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     5401 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5461 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     5521 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     5581 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     5641 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     5701 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
