LOCUS       sequence-446920-        5430 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0951h is an enhancer sequence, designed to drive AAV-mediated 
            transgene expression in striatal Pvalb+/Pthlh+ neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5430)
  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 5430)
  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: AiP13792 -
            pAAV-AiE0951h-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5430
                     /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      896..905
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(902..904,905..907,908..1618)
                     /codon_start=1
                     /product="monomeric optimized variant of EYFP, identical to
                     mVenus but with L68V (Kremers et al., 2006)"
                     /label=SYFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1208..1227)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1555..1576
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1628..1684
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1697..1762
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1793..2170
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(2257..2277)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2464..2575
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2693..2833
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2693..2822
                     /label=AAV2 ITR
     rep_origin      2908..3363
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2995..3014)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3205..3226
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3380..3399)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3499..3521
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3559..3577)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3645..3749
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3750..3818,3819..4610)
                     /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(3968..3987)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4781..5369
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5270..5289
                     /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 gcgttcattt tctttcccac agggtggtca gaaccaccct
      181 gggttcagcc cagctccctc tctgggacac atgtatcctt ggaggcctgt agccaccact
      241 cactcctctc agtgccctgc agactataca caaagcagga gtcaatgcac aataatggag
      301 atggggtgat taattgaggt gagggtgggt ggaacaagag gtcccaggtc ttgtgcttgg
      361 cacgaactct ctggttctgc ctctgccaat gtgtgctgca gacagctaag ctgacactgg
      421 catcacctgc actgagtctc tgtgtcatga ggtagtcatg gaaactcact ccttcctaag
      481 gtaatgagga gacaattagg tagaaatgag aaatcgagtt aattgcttgt catcccctct
      541 ctgctctctg tgtatttatt caaattaagc tttttcagtt ttctagtgtc agcagatacc
      601 aatgatcgga gcacaggctt gggagccagg acctccagga aacagccagt actgcatgaa
      661 tcacttgccc ttcctgcctc tttgtttagt cctgacccaa gcattcagga aaacaaatct
      721 ctttgcctga cctcccttgg cttctgaccg aaagtctctg aactgactta attttaacca
      781 tctctggctc cacatgcatt cagagctcgg gctgggcata aaagtcaggg cagagccatc
      841 tattgcttac atttgcttct gggatccaga tctttcgaag ctagcgctac cggtcgccac
      901 catggtctcc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
      961 cggcgatgtc aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     1021 cggcaagctg accctgaagc tgatctgcac caccggcaag ctgcccgtgc cctggcccac
     1081 cctcgtgacc accctgggct acggcgtgca gtgcttcgcc cgctaccccg accacatgaa
     1141 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     1201 cttcaaagac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     1261 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     1321 caagctggag tacaactaca acagccacaa cgtctatatc accgccgaca agcagaagaa
     1381 cggcatcaag gccaacttca agatccgcca caacatcgag gacggcggcg tgcagctcgc
     1441 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     1501 ctacctgagc taccagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt
     1561 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaaggg
     1621 ctctggtgct accaacttct cactgttgaa acaggcaggg gatgtagagg agaatccagg
     1681 gcctggtgct agtggagact acaaagacca tgacggagat tataaagatc atgacatcga
     1741 ttacaaggat gacgatgaca agtccggact cagatctgga ggctccggag gcccagagcc
     1801 agcgaagtct gctcccgccc cgaaaaaggg ctccaagaag gcggtgacta aggcgcagaa
     1861 gaaaggcggc aagaagcgca agcgcagccg caaggagagc tattccatct atgtgtacaa
     1921 ggttctgaag caggtccacc ctgacaccgg catttcgtcc aaggccatgg gcatcatgaa
     1981 ttcgtttgtg aacgacattt tcgagcgcat cgcaggagag gcttcccgcc tggcgcatta
     2041 caacaagcgc tcgaccatca cctcccggga gatccagacg gccgtgcgcc tgctgctgcc
     2101 tggggagttg gccaagcacg ccgtgtccga gggtactaag gccatcacca agtacaccag
     2161 cgctaagtaa tgaggcgcgc cgcggccgcg aattcgatat cataatcaac ctctggatta
     2221 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     2281 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     2341 ctccttgtat aaatcctggt tagttcttgc cacggcggaa ctcatcgccg cctgccttgc
     2401 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggctc gagagatctt
     2461 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     2521 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     2581 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     2641 attgggaaga caatagcagg catgagatct cacgtgcgga ccgagcggcc gcaggaaccc
     2701 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
     2761 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc
     2821 agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     2881 caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg
     2941 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt
     3001 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     3061 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     3121 atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     3181 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact
     3241 ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa
     3301 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
     3361 ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     3421 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     3481 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     3541 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     3601 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     3661 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     3721 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     3781 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     3841 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     3901 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     3961 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     4021 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     4081 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     4141 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     4201 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     4261 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     4321 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     4381 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     4441 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     4501 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     4561 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     4621 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     4681 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     4741 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     4801 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     4861 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     4921 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     4981 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     5041 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     5101 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     5161 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     5221 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     5281 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     5341 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     5401 tggccttttg ctggcctttt gctcacatgt
//
