LOCUS       sequence-446902-        5758 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0140h_3xC2 is an enhancer sequence, designed to drive 
            AAV-mediated transgene expression in striatal Pvalb+ neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5758)
  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 5758)
  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: AiP12577 -
            pAAV-AiE0140h_3xC2-minBG-ChR2(CRC)-...
FEATURES             Location/Qualifiers
     source          1..5758
                     /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      834..843
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             840..1766
                     /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
                     PSMLYLATGHRVQWLRYAEWLLTCPVICIRLSNLTGLSNDYSRRTMGLLVSDIGCIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     regulatory      1776..1785
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1782..1784,1785..1787,1788..2501)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1827..1848)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2088..2107)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2435..2456
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2585..2605)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2792..2903
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3021..3161
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3021..3150
                     /label=AAV2 ITR
     rep_origin      3236..3691
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3323..3342)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3533..3554
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3708..3727)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3827..3849
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3887..3905)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3973..4077
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4078..4146,4147..4938)
                     /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(4296..4315)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5109..5697
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5598..5617
                     /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 gcgtccaaga aattagaata aagggaaaaa tgactttgag
      181 atgtactttc ttaaaataac tgaagtaatg accgcatgag ccagctgatg ggtttttaaa
      241 tggatacgtt tctatcagcc tgctgcttga tgccagagcc aaatatatat ggagtgtatt
      301 atatcatatc ctgtaggcag gagactgttg gggcattggg ggacccaaga aattagaata
      361 aagggaaaaa tgactttgag atgtactttc ttaaaataac tgaagtaatg accgcatgag
      421 ccagctgatg ggtttttaaa tggatacgtt tctatcagcc tgctgcttga tgccagagcc
      481 aaatatatat ggagtgtatt atatcatatc ctgtaggcag gagactgttg gggcattggg
      541 ggacccaaga aattagaata aagggaaaaa tgactttgag atgtactttc ttaaaataac
      601 tgaagtaatg accgcatgag ccagctgatg ggtttttaaa tggatacgtt tctatcagcc
      661 tgctgcttga tgccagagcc aaatatatat ggagtgtatt atatcatatc ctgtaggcag
      721 gagactgttg gggcattggg ggacgagctc gggctgggca taaaagtcag ggcagagcca
      781 tctattgctt acatttgctt ctgggatcca gatctttcga agctagcgcg gccgccacca
      841 tggactatgg cggcgctttg tctgccgtcg gacgcgaact tttgttcgtt actaatcctg
      901 tggtggtgaa cgggtccgtc ctggtccctg aggatcaatg ttactgtgcc ggatggattg
      961 aatctcgcgg cacgaacggc gctcagaccg cgtcaaatgt cctgcagtgg cttgcagcag
     1021 gattcagcat tttgctgctg atgttctatg cctaccaaac ctggaaatct acatgcggct
     1081 gggaggagat ctatgtgtgc gccattgaaa tggttaaggt gattctcgag ttcttttttg
     1141 agtttaagaa tccctctatg ctctaccttg ccacaggaca ccgggtgcag tggctgcgct
     1201 atgcagagtg gctgctcact tgtcctgtca tctgcatccg cctgagcaac ctcaccggcc
     1261 tgagcaacga ctacagcagg agaaccatgg gactccttgt ctcagacatc gggtgtatcg
     1321 tgtggggggc taccagcgcc atggcaaccg gctatgttaa agtcatcttc ttttgtcttg
     1381 gattgtgcta tggcgcgaac acattttttc acgccgccaa agcatatatc gagggttatc
     1441 atactgtgcc aaagggtcgg tgccgccagg tcgtgaccgg catggcatgg ctgtttttcg
     1501 tgagctgggg tatgttccca attctcttca ttttggggcc cgaaggtttt ggcgtcctga
     1561 gcgtctatgg ctccaccgta ggtcacacga ttattgatct gatgagtaaa aattgttggg
     1621 ggttgttggg acactacctg cgcgtcctga tccacgagca catattgatt cacggagata
     1681 tccgcaaaac caccaaactg aacatcggcg gaacggagat cgaggtcgag actctcgtcg
     1741 aagacgaagc cgaggccgga gccgtgccag cggccgccac catggtgagc aagggcgagg
     1801 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     1861 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     1921 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accttcggct
     1981 acggcctgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     2041 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     2101 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     2161 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     2221 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     2281 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     2341 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc taccagtccg
     2401 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     2461 ccgccgggat cactctcggc atggacgagc tgtacaagta aggcgcgccg cggccgcgaa
     2521 ttcgatatca taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     2581 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     2641 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggtta gttcttgcca
     2701 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     2761 ctgacaattc cgtggctcga gagatcttcg actgtgcctt ctagttgcca gccatctgtt
     2821 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     2881 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     2941 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgagatctca
     3001 cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg
     3061 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
     3121 ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt
     3181 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg
     3241 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3301 cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3361 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     3421 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg
     3481 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3541 ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga tttataaggg
     3601 attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     3661 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct
     3721 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     3781 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     3841 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     3901 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     3961 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4021 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     4081 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     4141 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     4201 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     4261 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     4321 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     4381 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     4441 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     4501 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     4561 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     4621 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     4681 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     4741 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     4801 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     4861 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     4921 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     4981 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5041 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5101 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5161 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     5221 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     5281 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     5341 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     5401 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     5461 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     5521 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     5581 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     5641 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     5701 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt
//
