LOCUS       sequence-425169-        5802 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 5802)
  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 5802)
  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: AiP14036:
            pAAV-AiE0743m_3xC2-minBG-CoChR-EGFP-...
FEATURES             Location/Qualifiers
     source          1..5802
                     /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             963..1826
                     /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(1845..1847,1848..2561)
                     /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(1845..1847,1848..2558)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1887..1908)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2148..2167)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2495..2516
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2629..2649)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2836..2947
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3065..3205
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3065..3194
                     /label=AAV2 ITR
     rep_origin      3280..3735
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3367..3386)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3577..3598
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3752..3771)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3871..3893
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3931..3949)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4017..4121
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4122..4190,4191..4982)
                     /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(4340..4359)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5153..5741
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5642..5661
                     /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 ggatccgcca
      961 ccatgctggg aaacggcagc gccattgtgc ctatcgacca gtgcttttgc ctggcttgga
     1021 ccgacagcct gggaagcgat acagagcagc tggtggccaa catcctccag tggttcgcct
     1081 tcggcttcag catcctgatc ctgatgttct acgcctacca gacttggaga gccacttgcg
     1141 gttgggagga ggtctacgtc tgttgcgtcg agctgaccaa ggtcatcatc gagttcttcc
     1201 acgagttcga cgaccccagc atgctgtacc tggctaacgg acaccgagtc cagtggctga
     1261 gatacgcaga gtggctgctg acttgtcccg tcatcctgat ccacctgagc aacctgaccg
     1321 gcctgaagga cgactacagc aagcggacca tgaggctgct ggtgtcagac gtgggaacca
     1381 tcgtgtgggg agctacaagc gccatgagca caggctacgt caaggtcatc ttcttcgtgc
     1441 tgggttgcat ctacggcgcc aacaccttct tccacgccgc caaggtgtat atcgagagct
     1501 accacgtggt gccaaagggc agacctagaa ccgtcgtgcg gatcatggct tggctgttct
     1561 tcctgtcttg gggcatgttc cccgtgctgt tcgtcgtggg accagaagga ttcgacgcca
     1621 tcagcgtgta cggctctacc attggccaca ccatcatcga cctcatgagc aagaattgtt
     1681 ggggcctgct gggacactat ctgagagtgc tgatccacca gcacatcatc atctacggcg
     1741 acatccggaa gaagaccaag atcaacgtgg ccggcgagga gatggaagtg gagaccatgg
     1801 tggaccagga ggacgaggag acagtggcgg caccggtagt agcagtgagc aagggcgagg
     1861 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     1921 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     1981 tcatttgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     2041 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     2101 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     2161 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     2221 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     2281 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     2341 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     2401 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     2461 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     2521 ccgccgggat cactctcggc atggacgagc tgtacaagta agaattcgat atcataatca
     2581 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     2641 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     2701 tttcattttc tcctccttgt ataaatcctg gttagttctt gccacggcgg aactcatcgc
     2761 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggc
     2821 tcgagagatc ttcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     2881 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     2941 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     3001 gcaaggggga ggattgggaa gacaatagca ggcatgagat ctcacgtgcg gaccgagcgg
     3061 ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     3121 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
     3181 gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg
     3241 tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat
     3301 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag
     3361 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     3421 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     3481 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     3541 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     3601 caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
     3661 tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     3721 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     3781 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     3841 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     3901 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     3961 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     4021 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     4081 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     4141 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     4201 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     4261 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     4321 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     4381 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     4441 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     4501 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     4561 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     4621 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     4681 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     4741 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     4801 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     4861 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     4921 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     4981 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     5041 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     5101 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     5161 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     5221 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     5281 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     5341 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5401 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     5461 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     5521 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     5581 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     5641 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     5701 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     5761 ttttacggtt cctggccttt tgctggcctt ttgctcacat gt
//
