LOCUS       sequence-424977-        5900 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ChR2(H134R)-EYFP in striatal cholinergic neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5900)
  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 5900)
  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: AiP13755:
            pAAV-AiE0743m_3xC2-minBG-ChR2(H134R)...
FEATURES             Location/Qualifiers
     source          1..5900
                     /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      973..982
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             979..1905
                     /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      1915..1924
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1921..1923,1924..1926,1927..2640)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1966..1987)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2227..2246)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2574..2595
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2727..2747)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2934..3045
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3163..3303
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3163..3292
                     /label=AAV2 ITR
     rep_origin      3378..3833
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3465..3484)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3675..3696
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3850..3869)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3969..3991
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4029..4047)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4115..4219
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4220..4288,4289..5080)
                     /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(4438..4457)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5251..5839
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5740..5759
                     /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 gcgtagcagc agctagtagt acatacaaag aaatagaaaa
      181 catgaagacc aggggagttt aattatatgg ttgcaaaaat ggttcaacat tgcaaagttg
      241 ttaatactca gtggatgggg agggagctgt gcaattatct ccacaggtac agaaaaggca
      301 cttagtaaaa tgtagccaac tctttggctt ctaaaaaagg agaaaatggt ttacaaaaac
      361 aagcctccct ggatgcttcc ttaatttgtt cttagcagca gctagtagta catacaaaga
      421 aatagaaaac atgaagacca ggggagttta attatatggt tgcaaaaatg gttcaacatt
      481 gcaaagttgt taatactcag tggatgggga gggagctgtg caattatctc cacaggtaca
      541 gaaaaggcac ttagtaaaat gtagccaact ctttggcttc taaaaaagga gaaaatggtt
      601 tacaaaaaca agcctccctg gatgcttcct taatttgttc ttagcagcag ctagtagtac
      661 atacaaagaa atagaaaaca tgaagaccag gggagtttaa ttatatggtt gcaaaaatgg
      721 ttcaacattg caaagttgtt aatactcagt ggatggggag ggagctgtgc aattatctcc
      781 acaggtacag aaaaggcact tagtaaaatg tagccaactc tttggcttct aaaaaaggag
      841 aaaatggttt acaaaaacaa gcctccctgg atgcttcctt aatttgttct tgagctcggg
      901 ctgggcataa aagtcagggc agagccatct attgcttaca tttgcttctg ggatccagat
      961 ctttcgaagc tagccaccat ggactatggc ggcgctttgt ctgccgtcgg acgcgaactt
     1021 ttgttcgtta ctaatcctgt ggtggtgaac gggtccgtcc tggtccctga ggatcaatgt
     1081 tactgtgccg gatggattga atctcgcggc acgaacggcg ctcagaccgc gtcaaatgtc
     1141 ctgcagtggc ttgcagcagg attcagcatt ttgctgctga tgttctatgc ctaccaaacc
     1201 tggaaatcta catgcggctg ggaggagatc tatgtgtgcg ccattgaaat ggttaaggtg
     1261 attctcgagt tcttttttga gtttaagaat ccctctatgc tctaccttgc cacaggacac
     1321 cgggtgcagt ggctgcgcta tgcagagtgg ctgctcactt gtcctgtcat ccttatccgc
     1381 ctgagcaacc tcaccggcct gagcaacgac tacagcagga gaaccatggg actccttgtc
     1441 tcagacatcg ggactatcgt gtggggggct accagcgcca tggcaaccgg ctatgttaaa
     1501 gtcatcttct tttgtcttgg attgtgctat ggcgcgaaca cattttttca cgccgccaaa
     1561 gcatatatcg agggttatca tactgtgcca aagggtcggt gccgccaggt cgtgaccggc
     1621 atggcatggc tgtttttcgt gagctggggt atgttcccaa ttctcttcat tttggggccc
     1681 gaaggttttg gcgtcctgag cgtctatggc tccaccgtag gtcacacgat tattgatctg
     1741 atgagtaaaa attgttgggg gttgttggga cactacctgc gcgtcctgat ccacgagcac
     1801 atattgattc acggagatat ccgcaaaacc accaaactga acatcggcgg aacggagatc
     1861 gaggtcgaga ctctcgtcga agacgaagcc gaggccggag ccgtgccagc ggccgccacc
     1921 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     1981 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     2041 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     2101 ctcgtgacca ccttcggcta cggcctgcag tgcttcgccc gctaccccga ccacatgaag
     2161 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     2221 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     2281 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     2341 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     2401 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     2461 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     2521 tacctgagct accagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     2581 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
     2641 tagggcgcgc cgcggccgcg aattcgatat cataatcaac ctctggatta caaaatttgt
     2701 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     2761 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     2821 aaatcctggt tagttcttgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     2881 acaggggctc ggctgttggg cactgacaat tccgtggctc gagagatctt cgactgtgcc
     2941 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     3001 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     3061 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     3121 caatagcagg catgagatct cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg
     3181 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
     3241 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc
     3301 aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
     3361 gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     3421 tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt
     3481 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     3541 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga
     3601 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     3661 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg
     3721 ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct
     3781 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg
     3841 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     3901 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     3961 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     4021 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     4081 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     4141 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     4201 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     4261 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     4321 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     4381 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     4441 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     4501 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     4561 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     4621 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     4681 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     4741 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     4801 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     4861 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     4921 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     4981 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     5041 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     5101 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     5161 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     5221 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5281 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5341 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     5401 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     5461 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5521 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     5581 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     5641 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     5701 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     5761 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     5821 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     5881 ctggcctttt gctcacatgt
//
