LOCUS       sequence-430971-        5964 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of CoChR-EGFP in striatal cholinergic neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5964)
  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 5964)
  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: AiP15140:
            pAAV-AiE0873m_3xC2-minBG-CoChR-EGFP-...
FEATURES             Location/Qualifiers
     source          1..5964
                     /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             1125..1988
                     /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(2007..2009,2010..2723)
                     /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(2007..2009,2010..2720)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2049..2070)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2310..2329)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2657..2678
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2791..2811)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2998..3109
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3227..3367
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3227..3356
                     /label=AAV2 ITR
     rep_origin      3442..3897
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3529..3548)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3739..3760
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3914..3933)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4033..4055
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4093..4111)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4179..4283
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4284..4352,4353..5144)
                     /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(4502..4521)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5315..5903
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5804..5823
                     /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 gcgtcatgga tttctccaaa gctttgtagg gcagaactct
      181 gaatgtttat ttctctgatg aatccttaat gaaagtggcc agccatcctg gggagaacaa
      241 gccttagttc ccactgagaa ggatagcaag cagcataaga gtagctgatg tcaaaggcta
      301 ttaatgcaaa tgaaaattat aattcctcat caagatctgc tcccagacag gctaatcatt
      361 gtgccacctg tttacagaaa cacactgcta atcagtcagt aaaatctttt aagtttctgt
      421 gcttctctgt gtacgatttt tagcctccat ggatttctcc aaagctttgt agggcagaac
      481 tctgaatgtt tatttctctg atgaatcctt aatgaaagtg gccagccatc ctggggagaa
      541 caagccttag ttcccactga gaaggatagc aagcagcata agagtagctg atgtcaaagg
      601 ctattaatgc aaatgaaaat tataattcct catcaagatc tgctcccaga caggctaatc
      661 attgtgccac ctgtttacag aaacacactg ctaatcagtc agtaaaatct tttaagtttc
      721 tgtgcttctc tgtgtacgat ttttagcctc catggatttc tccaaagctt tgtagggcag
      781 aactctgaat gtttatttct ctgatgaatc cttaatgaaa gtggccagcc atcctgggga
      841 gaacaagcct tagttcccac tgagaaggat agcaagcagc ataagagtag ctgatgtcaa
      901 aggctattaa tgcaaatgaa aattataatt cctcatcaag atctgctccc agacaggcta
      961 atcattgtgc cacctgttta cagaaacaca ctgctaatca gtcagtaaaa tcttttaagt
     1021 ttctgtgctt ctctgtgtac gatttttagc ctcgagctcg ggctgggcat aaaagtcagg
     1081 gcagagccat ctattgctta catttgcttc tgggatccgc caccatgctg ggaaacggca
     1141 gcgccattgt gcctatcgac cagtgctttt gcctggcttg gaccgacagc ctgggaagcg
     1201 atacagagca gctggtggcc aacatcctcc agtggttcgc cttcggcttc agcatcctga
     1261 tcctgatgtt ctacgcctac cagacttgga gagccacttg cggttgggag gaggtctacg
     1321 tctgttgcgt cgagctgacc aaggtcatca tcgagttctt ccacgagttc gacgacccca
     1381 gcatgctgta cctggctaac ggacaccgag tccagtggct gagatacgca gagtggctgc
     1441 tgacttgtcc cgtcatcctg atccacctga gcaacctgac cggcctgaag gacgactaca
     1501 gcaagcggac catgaggctg ctggtgtcag acgtgggaac catcgtgtgg ggagctacaa
     1561 gcgccatgag cacaggctac gtcaaggtca tcttcttcgt gctgggttgc atctacggcg
     1621 ccaacacctt cttccacgcc gccaaggtgt atatcgagag ctaccacgtg gtgccaaagg
     1681 gcagacctag aaccgtcgtg cggatcatgg cttggctgtt cttcctgtct tggggcatgt
     1741 tccccgtgct gttcgtcgtg ggaccagaag gattcgacgc catcagcgtg tacggctcta
     1801 ccattggcca caccatcatc gacctcatga gcaagaattg ttggggcctg ctgggacact
     1861 atctgagagt gctgatccac cagcacatca tcatctacgg cgacatccgg aagaagacca
     1921 agatcaacgt ggccggcgag gagatggaag tggagaccat ggtggaccag gaggacgagg
     1981 agacagtggc ggcaccggta gtagcagtga gcaagggcga ggagctgttc accggggtgg
     2041 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     2101 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatttgc accaccggca
     2161 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
     2221 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     2281 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     2341 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     2401 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     2461 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
     2521 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     2581 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
     2641 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     2701 gcatggacga gctgtacaag taagaattcg atatcataat caacctctgg attacaaaat
     2761 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     2821 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     2881 gtataaatcc tggttagttc ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     2941 ctggacaggg gctcggctgt tgggcactga caattccgtg gctcgagaga tcttcgactg
     3001 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     3061 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     3121 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     3181 aagacaatag caggcatgag atctcacgtg cggaccgagc ggccgcagga acccctagtg
     3241 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag
     3301 gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc
     3361 ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     3421 atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     3481 tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt
     3541 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     3601 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg
     3661 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     3721 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct
     3781 cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg
     3841 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat
     3901 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     3961 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     4021 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     4081 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     4141 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     4201 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     4261 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     4321 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     4381 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     4441 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     4501 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     4561 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     4621 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     4681 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     4741 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     4801 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     4861 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     4921 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     4981 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     5041 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     5101 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     5161 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5221 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5281 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5341 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5401 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     5461 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5521 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5581 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     5641 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     5701 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     5761 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     5821 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     5881 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     5941 tttgctggcc ttttgctcac atgt
//
