LOCUS       sequence-472646-        5320 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0140h is an enhancer sequence, designed to drive AAV-mediated 
            transgene expression in cortical and hippocampal Pvalb neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5320)
  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 5320)
  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: AiP13401: pAAV-AiE0140h-minBG-FlpO-4X2C-WPRE3-...
FEATURES             Location/Qualifiers
     source          1..5320
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1497..1517)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1704..1815
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1933..2073
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1933..2062
                     /label=AAV2 ITR
     rep_origin      2148..2603
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2235..2254)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2445..2466
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2620..2639)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2739..2761
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2799..2817)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2885..2989
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2990..3058,3059..3850)
                     /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(3208..3227)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4021..4609
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4510..4529
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4671..4800
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      5203..5212
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5209..5214,5215..5235,5236..5320,1..1187)
                     /codon_start=1
                     /product="nuclear-targeted site-specific recombinase"
                     /label=FLPo
                     /note="FLPo is a mouse codon-optimized fusion between the 
                     SV40 nuclear localization signal and a thermostable version
                     of the Saccharomyces cerevisiae site-specific recombinase 
                     FLP.  Recombination occurs at FRT sequences."
                     /translation="MAPKKKRKVMSQFDILCKTPPKVLVRQFVERFERPSGEKIASCAA
                     ELTYLCWMITHNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKK
                     LIPAWEFTIIPYNGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESI
                     WEITEKILNSFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGV
                     IIQCLVTETKTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEY
                     QLLKDNLVRSYNKALKKNAPYPIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWS
                     DKRASAVARTTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQL
                     KGSAEGSIRYPAWNGIISQEVLDYLSSYINRRI"
ORIGIN
        1 agaagatcgc cagctgtgcc gccgagctga cctacctgtg ctggatgatc acccacaacg
       61 gcaccgccat caagagggcc accttcatga gctacaacac catcatcagc aacagcctga
      121 gcttcgacat cgtgaacaag agcctgcagt tcaagtacaa gacccagaag gccaccatcc
      181 tggaggccag cctgaagaag ctgatccccg cctgggagtt caccatcatc ccttacaacg
      241 gccagaagca ccagagcgac atcaccgaca tcgtgtccag cctgcagctg cagttcgaga
      301 gcagcgagga ggccgacaag ggcaacagcc acagcaagaa gatgctgaag gccctgctgt
      361 ccgagggcga gagcatctgg gagatcaccg agaagatcct gaacagcttc gagtacacca
      421 gcaggttcac caagaccaag accctgtacc agttcctgtt cctggccaca ttcatcaact
      481 gcggcaggtt cagcgacatc aagaacgtgg accccaagag cttcaagctg gtgcagaaca
      541 agtacctggg cgtgatcatt cagtgcctgg tgaccgagac caagacaagc gtgtccaggc
      601 acatctactt tttcagcgcc agaggcagga tcgaccccct ggtgtacctg gacgagttcc
      661 tgaggaacag cgagcccgtg ctgaagagag tgaacaggac cggcaacagc agcagcaaca
      721 agcaggagta ccagctgctg aaggacaacc tggtgcgcag ctacaacaag gccctgaaga
      781 agaacgcccc ctaccccatc ttcgctatca agaacggccc taagagccac atcggcaggc
      841 acctgatgac cagctttctg agcatgaagg gcctgaccga gctgacaaac gtggtgggca
      901 actggagcga caagagggcc tccgccgtgg ccaggaccac ctacacccac cagatcaccg
      961 ccatccccga ccactacttc gccctggtgt ccaggtacta cgcctacgac cccatcagca
     1021 aggagatgat cgccctgaag gacgagacca accccatcga ggagtggcag cacatcgagc
     1081 agctgaaggg cagcgccgag ggcagcatca gataccccgc ctggaacggc atcatcagcc
     1141 aggaggtgct ggactacctg agcagctaca tcaacaggcg gatctgagtc gacggcgcgc
     1201 cgcggcctta aagagaccgg ttcactgtga cagtaaaaga gaccggttca ctgtgagaat
     1261 gaaagagacc ggttcactgt gatcggaaaa gagaccggtt cactgtgagc ggccttgaaa
     1321 cccagcagac aatgtagctc agtagaaacc cagcagacaa tgtagctgaa tggaaaccca
     1381 gcagacaatg tagcttcgga gaaacccagc agacaatgta gctgtcgacg aattcgatat
     1441 cataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     1501 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     1561 cgtatggctt tcattttctc ctccttgtat aaatcctggt tagttcttgc cacggcggaa
     1621 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     1681 tccgtggctc gagagatctt cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     1741 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     1801 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     1861 gcaggacagc aagggggagg attgggaaga caatagcagg catgagatct cacgtgcgga
     1921 ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     1981 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     2041 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta
     2101 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag
     2161 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     2221 cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     2281 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     2341 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata
     2401 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     2461 aactggaaca acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc
     2521 gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     2581 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc
     2641 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     2701 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     2761 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     2821 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     2881 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     2941 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     3001 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     3061 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     3121 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     3181 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     3241 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     3301 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     3361 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     3421 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     3481 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     3541 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     3601 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     3661 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     3721 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     3781 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     3841 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     3901 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     3961 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     4021 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     4081 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     4141 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     4201 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     4261 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     4321 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     4381 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     4441 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     4501 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     4561 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     4621 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt cctgcaggca
     4681 gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt ggtcgcccgg
     4741 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
     4801 gcggccgcac gcgtggttgt gtgtgaagat taagcccttt cttgctctat gtatttgagc
     4861 ctagaaaaag acacactccc aagaaattag aataaaggga aaaatgactt tgagatgtac
     4921 tttcttaaaa taactgaagt aatgaccgca tgagccagct gatgggtttt taaatggata
     4981 cgtttctatc agcctgctgc ttgatgccag agccaaatat atatggagtg tattatatca
     5041 tatcctgtag gcaggagact gttggggcat tgggggactt agagaggtga agtggcaggc
     5101 ttggcacagg aattaacagc agagctcggg ctgggcataa aagtcagggc agagccatct
     5161 attgcttaca tttgcttctg ggatccagat ctttcgaagc tagccaccat ggctcctaag
     5221 aagaagagga aggtgatgag ccagttcgac atcctgtgca agaccccccc caaggtgctg
     5281 gtgcggcagt tcgtggagag attcgagagg cccagcggcg
//
