LOCUS       sequence-459328-        5277 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0445h_3xC2 is an enhancer sequence, designed to drive 
            AAV-mediated transgene expression in striatal indirect pathway MSNs 
            (D2-MSNs).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5277)
  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 5277)
  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: AiP14034:
            pAAV-AiE0445h_3xC2-minBG-CoChR-EGFP-...
FEATURES             Location/Qualifiers
     source          1..5277
                     /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             438..1301
                     /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(1320..1322,1323..2036)
                     /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(1320..1322,1323..2033)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1362..1383)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1623..1642)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1970..1991
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2104..2124)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2311..2422
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2540..2680
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2540..2669
                     /label=AAV2 ITR
     rep_origin      2755..3210
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2842..2861)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3052..3073
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3227..3246)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3346..3368
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3406..3424)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3492..3596
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3597..3665,3666..4457)
                     /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(3815..3834)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4628..5216
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5117..5136
                     /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 gcgtgtggag gactttgagc ttaaccctat taaatttaat
      181 ctaatatatt cagcagcctg tcacaatctt gttaaatctt tattccgtca cccaccttat
      241 tagggggccc tcccacttta gtcatcagca gattagtaag catgccattt agatgtcatc
      301 taagttaatg aacctgactg agccagagtc acatggcaga gtcacagaaa ctccttctga
      361 gctggcgagc tcgggctggg cataaaagtc agggcagagc catctattgc ttacatttgc
      421 ttctgggatc cgccaccatg ctgggaaacg gcagcgccat tgtgcctatc gaccagtgct
      481 tttgcctggc ttggaccgac agcctgggaa gcgatacaga gcagctggtg gccaacatcc
      541 tccagtggtt cgccttcggc ttcagcatcc tgatcctgat gttctacgcc taccagactt
      601 ggagagccac ttgcggttgg gaggaggtct acgtctgttg cgtcgagctg accaaggtca
      661 tcatcgagtt cttccacgag ttcgacgacc ccagcatgct gtacctggct aacggacacc
      721 gagtccagtg gctgagatac gcagagtggc tgctgacttg tcccgtcatc ctgatccacc
      781 tgagcaacct gaccggcctg aaggacgact acagcaagcg gaccatgagg ctgctggtgt
      841 cagacgtggg aaccatcgtg tggggagcta caagcgccat gagcacaggc tacgtcaagg
      901 tcatcttctt cgtgctgggt tgcatctacg gcgccaacac cttcttccac gccgccaagg
      961 tgtatatcga gagctaccac gtggtgccaa agggcagacc tagaaccgtc gtgcggatca
     1021 tggcttggct gttcttcctg tcttggggca tgttccccgt gctgttcgtc gtgggaccag
     1081 aaggattcga cgccatcagc gtgtacggct ctaccattgg ccacaccatc atcgacctca
     1141 tgagcaagaa ttgttggggc ctgctgggac actatctgag agtgctgatc caccagcaca
     1201 tcatcatcta cggcgacatc cggaagaaga ccaagatcaa cgtggccggc gaggagatgg
     1261 aagtggagac catggtggac caggaggacg aggagacagt ggcggcaccg gtagtagcag
     1321 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     1381 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     1441 agctgaccct gaagttcatt tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     1501 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     1561 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     1621 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     1681 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     1741 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     1801 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     1861 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     1921 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     1981 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaagaat
     2041 tcgatatcat aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     2101 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     2161 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttag ttcttgccac
     2221 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     2281 tgacaattcc gtggctcgag agatcttcga ctgtgccttc tagttgccag ccatctgttg
     2341 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     2401 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     2461 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gagatctcac
     2521 gtgcggaccg agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc
     2581 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg
     2641 gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt
     2701 ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc
     2761 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     2821 ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     2881 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     2941 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     3001 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     3061 tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga
     3121 ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     3181 attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg
     3241 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     3301 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     3361 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     3421 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     3481 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3541 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     3601 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     3661 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     3721 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     3781 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     3841 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     3901 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     3961 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     4021 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     4081 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     4141 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     4201 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     4261 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     4321 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     4381 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     4441 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     4501 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4561 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4621 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4681 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4741 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     4801 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4861 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4921 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4981 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     5041 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     5101 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     5161 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     5221 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgt
//
