LOCUS       sequence-450138-        5837 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0452h_3xC2 is an enhancer sequence, designed to drive 
            AAV-mediated transgene expression in striatal D2 MSNs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5837)
  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 5837)
  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: AiP14870 -
            pAAV-AiE0452h_3xC2-minBG-ChR2(H134R...
FEATURES             Location/Qualifiers
     source          1..5837
                     /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      910..919
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             916..1842
                     /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      1852..1861
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1858..1860,1861..1863,1864..2577)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1903..1924)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2164..2183)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2511..2532
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2664..2684)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2871..2982
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3100..3240
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3100..3229
                     /label=AAV2 ITR
     rep_origin      3315..3770
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3402..3421)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3612..3633
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3787..3806)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3906..3928
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3966..3984)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4052..4156
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4157..4225,4226..5017)
                     /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(4375..4394)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5188..5776
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5677..5696
                     /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 gcgtgtgaaa aatacctagt cttgaacttt ggagttccaa
      181 attccacatg atcgacttcc tcaaggcttt tcacttgacg aacaaattgc atgccaagaa
      241 atcgtcaaaa ttctatcaag cttaattatg caatttctgt ctataaatga catacctcct
      301 gtaccctatg acacccacct atgcactaat aagatgtgac agctgcatca ccaacaaagc
      361 ctcccccgcc tggtgaaaaa tacctagtct tgaactttgg agttccaaat tccacatgat
      421 cgacttcctc aaggcttttc acttgacgaa caaattgcat gccaagaaat cgtcaaaatt
      481 ctatcaagct taattatgca atttctgtct ataaatgaca tacctcctgt accctatgac
      541 acccacctat gcactaataa gatgtgacag ctgcatcacc aacaaagcct cccccgcctg
      601 gtgaaaaata cctagtcttg aactttggag ttccaaattc cacatgatcg acttcctcaa
      661 ggcttttcac ttgacgaaca aattgcatgc caagaaatcg tcaaaattct atcaagctta
      721 attatgcaat ttctgtctat aaatgacata cctcctgtac cctatgacac ccacctatgc
      781 actaataaga tgtgacagct gcatcaccaa caaagcctcc cccgcctgga gctcgggctg
      841 ggcataaaag tcagggcaga gccatctatt gcttacattt gcttctggga tccagatctt
      901 tcgaagctag ccaccatgga ctatggcggc gctttgtctg ccgtcggacg cgaacttttg
      961 ttcgttacta atcctgtggt ggtgaacggg tccgtcctgg tccctgagga tcaatgttac
     1021 tgtgccggat ggattgaatc tcgcggcacg aacggcgctc agaccgcgtc aaatgtcctg
     1081 cagtggcttg cagcaggatt cagcattttg ctgctgatgt tctatgccta ccaaacctgg
     1141 aaatctacat gcggctggga ggagatctat gtgtgcgcca ttgaaatggt taaggtgatt
     1201 ctcgagttct tttttgagtt taagaatccc tctatgctct accttgccac aggacaccgg
     1261 gtgcagtggc tgcgctatgc agagtggctg ctcacttgtc ctgtcatcct tatccgcctg
     1321 agcaacctca ccggcctgag caacgactac agcaggagaa ccatgggact ccttgtctca
     1381 gacatcggga ctatcgtgtg gggggctacc agcgccatgg caaccggcta tgttaaagtc
     1441 atcttctttt gtcttggatt gtgctatggc gcgaacacat tttttcacgc cgccaaagca
     1501 tatatcgagg gttatcatac tgtgccaaag ggtcggtgcc gccaggtcgt gaccggcatg
     1561 gcatggctgt ttttcgtgag ctggggtatg ttcccaattc tcttcatttt ggggcccgaa
     1621 ggttttggcg tcctgagcgt ctatggctcc accgtaggtc acacgattat tgatctgatg
     1681 agtaaaaatt gttgggggtt gttgggacac tacctgcgcg tcctgatcca cgagcacata
     1741 ttgattcacg gagatatccg caaaaccacc aaactgaaca tcggcggaac ggagatcgag
     1801 gtcgagactc tcgtcgaaga cgaagccgag gccggagccg tgccagcggc cgccaccatg
     1861 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     1921 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     1981 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     2041 gtgaccacct tcggctacgg cctgcagtgc ttcgcccgct accccgacca catgaagcag
     2101 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     2161 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     2221 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     2281 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     2341 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     2401 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     2461 ctgagctacc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     2521 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaatag
     2581 ggcgcgccgc ggccgcgaat tcgatatcat aatcaacctc tggattacaa aatttgtgaa
     2641 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     2701 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     2761 tcctggttag ttcttgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     2821 ggggctcggc tgttgggcac tgacaattcc gtggctcgag agatcttcga ctgtgccttc
     2881 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     2941 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     3001 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     3061 tagcaggcat gagatctcac gtgcggaccg agcggccgca ggaaccccta gtgatggagt
     3121 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     3181 gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg
     3241 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc
     3301 aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     3361 gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg ctttcttccc
     3421 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     3481 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg
     3541 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     3601 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta tctcgggcta
     3661 ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa atgagctgat
     3721 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac
     3781 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     3841 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     3901 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     3961 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     4021 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4081 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4141 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     4201 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     4261 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     4321 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     4381 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     4441 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     4501 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     4561 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     4621 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     4681 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     4741 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     4801 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     4861 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     4921 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     4981 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     5041 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5101 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5161 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5221 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     5281 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     5341 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5401 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     5461 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     5521 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     5581 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     5641 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     5701 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     5761 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     5821 gccttttgct cacatgt
//
