LOCUS       sequence-425084-        5637 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of CoChR-EGFP 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 5637)
  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 5637)
  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: AiP14033:
            pAAV-AiE0779m_3xC2-minBG-CoChR-EGFP-...
FEATURES             Location/Qualifiers
     source          1..5637
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             798..1661
                     /codon_start=1
                     /product="high-photocurrent channelrhodopsin from 
                     Chloromonas oogama (Shemesh et al., 2017)"
                     /label=CoChR
                     /translation="MLGNGSAIVPIDQCFCLAWTDSLGSDTEQLVANILQWFAFGFSIL
                     ILMFYAYQTWRATCGWEEVYVCCVELTKVIIEFFHEFDDPSMLYLANGHRVQWLRYAEW
                     LLTCPVILIHLSNLTGLKDDYSKRTMRLLVSDVGTIVWGATSAMSTGYVKVIFFVLGCI
                     YGANTFFHAAKVYIESYHVVPKGRPRTVVRIMAWLFFLSWGMFPVLFVVGPEGFDAISV
                     YGSTIGHTIIDLMSKNCWGLLGHYLRVLIHQHIIIYGDIRKKTKINVAGEEMEVETMVD
                     QEDEETV"
     CDS             join(1680..1682,1683..2396)
                     /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(1680..1682,1683..2393)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1722..1743)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1983..2002)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2330..2351
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2464..2484)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2671..2782
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2900..3040
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2900..3029
                     /label=AAV2 ITR
     rep_origin      3115..3570
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3202..3221)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3412..3433
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3587..3606)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3706..3728
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3766..3784)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3852..3956
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3957..4025,4026..4817)
                     /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(4175..4194)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4988..5576
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5477..5496
                     /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 cgccaccatg ctgggaaacg gcagcgccat tgtgcctatc gaccagtgct
      841 tttgcctggc ttggaccgac agcctgggaa gcgatacaga gcagctggtg gccaacatcc
      901 tccagtggtt cgccttcggc ttcagcatcc tgatcctgat gttctacgcc taccagactt
      961 ggagagccac ttgcggttgg gaggaggtct acgtctgttg cgtcgagctg accaaggtca
     1021 tcatcgagtt cttccacgag ttcgacgacc ccagcatgct gtacctggct aacggacacc
     1081 gagtccagtg gctgagatac gcagagtggc tgctgacttg tcccgtcatc ctgatccacc
     1141 tgagcaacct gaccggcctg aaggacgact acagcaagcg gaccatgagg ctgctggtgt
     1201 cagacgtggg aaccatcgtg tggggagcta caagcgccat gagcacaggc tacgtcaagg
     1261 tcatcttctt cgtgctgggt tgcatctacg gcgccaacac cttcttccac gccgccaagg
     1321 tgtatatcga gagctaccac gtggtgccaa agggcagacc tagaaccgtc gtgcggatca
     1381 tggcttggct gttcttcctg tcttggggca tgttccccgt gctgttcgtc gtgggaccag
     1441 aaggattcga cgccatcagc gtgtacggct ctaccattgg ccacaccatc atcgacctca
     1501 tgagcaagaa ttgttggggc ctgctgggac actatctgag agtgctgatc caccagcaca
     1561 tcatcatcta cggcgacatc cggaagaaga ccaagatcaa cgtggccggc gaggagatgg
     1621 aagtggagac catggtggac caggaggacg aggagacagt ggcggcaccg gtagtagcag
     1681 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     1741 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     1801 agctgaccct gaagttcatt tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     1861 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     1921 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     1981 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     2041 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     2101 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     2161 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     2221 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     2281 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     2341 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaagaat
     2401 tcgatatcat aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     2461 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     2521 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttag ttcttgccac
     2581 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     2641 tgacaattcc gtggctcgag agatcttcga ctgtgccttc tagttgccag ccatctgttg
     2701 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     2761 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     2821 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gagatctcac
     2881 gtgcggaccg agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc
     2941 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg
     3001 gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt
     3061 ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc
     3121 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3181 ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3241 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     3301 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     3361 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     3421 tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga
     3481 ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     3541 attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg
     3601 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     3661 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     3721 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     3781 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     3841 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3901 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     3961 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     4021 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     4081 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     4141 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     4201 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     4261 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     4321 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     4381 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     4441 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     4501 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     4561 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     4621 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     4681 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     4741 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     4801 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     4861 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4921 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4981 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     5041 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     5101 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     5161 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     5221 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     5281 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     5341 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     5401 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     5461 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     5521 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     5581 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgt
//
