LOCUS       sequence-459479-        5379 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0452h 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 5379)
  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 5379)
  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: AiP14573: pAAV-AiE0452h-minBG-FlpO-WPRE3-BGHpA...
FEATURES             Location/Qualifiers
     source          1..5379
                     /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      812..821
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(818..823,824..844,845..2116)
                     /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"
     primer_bind     complement(2206..2226)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2413..2524
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2642..2782
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2642..2771
                     /label=AAV2 ITR
     rep_origin      2857..3312
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2944..2963)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3154..3175
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3329..3348)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3448..3470
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3508..3526)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3594..3698
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3699..3767,3768..4559)
                     /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(3917..3936)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4730..5318
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5219..5238
                     /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 gcgtagggag gaactaaagt ccattcatat gtccatattc
      181 tcaatttggg aaaaaatgtg gctttgtctt catcctcttt tcctgagtgt ataaggccag
      241 aagccaaagg caagagcagg acattccagg agcctttatg attgctctaa gtcacatctg
      301 ctctcgagct cccattccac tgtgaaaaat acctagtctt gaactttgga gttccaaatt
      361 ccacatgatc gacttcctca aggcttttca cttgacgaac aaattgcatg ccaagaaatc
      421 gtcaaaattc tatcaagctt aattatgcaa tttctgtcta taaatgacat acctcctgta
      481 ccctatgaca cccacctatg cactaataag atgtgacagc tgcatcacca acaaagcctc
      541 ccccgcctgc cattctcttc ggaggtgagc acctcctgct cgggaggaac tgacagcccc
      601 tttcagaaat gtgaaggcag tgaagtccca aaccagaaac agtgtgacca cctaggctca
      661 cccaaggacc gtcctgtgct tctgggcaac caatggatgt agggggttag gaggcagtgg
      721 ggattcgtca gagctcgggc tgggcataaa agtcagggca gagccatcta ttgcttacat
      781 ttgcttctgg gatccagatc tttcgaagct agccaccatg gctcctaaga agaagaggaa
      841 ggtgatgagc cagttcgaca tcctgtgcaa gacccccccc aaggtgctgg tgcggcagtt
      901 cgtggagaga ttcgagaggc ccagcggcga gaagatcgcc agctgtgccg ccgagctgac
      961 ctacctgtgc tggatgatca cccacaacgg caccgccatc aagagggcca ccttcatgag
     1021 ctacaacacc atcatcagca acagcctgag cttcgacatc gtgaacaaga gcctgcagtt
     1081 caagtacaag acccagaagg ccaccatcct ggaggccagc ctgaagaagc tgatccccgc
     1141 ctgggagttc accatcatcc cttacaacgg ccagaagcac cagagcgaca tcaccgacat
     1201 cgtgtccagc ctgcagctgc agttcgagag cagcgaggag gccgacaagg gcaacagcca
     1261 cagcaagaag atgctgaagg ccctgctgtc cgagggcgag agcatctggg agatcaccga
     1321 gaagatcctg aacagcttcg agtacaccag caggttcacc aagaccaaga ccctgtacca
     1381 gttcctgttc ctggccacat tcatcaactg cggcaggttc agcgacatca agaacgtgga
     1441 ccccaagagc ttcaagctgg tgcagaacaa gtacctgggc gtgatcattc agtgcctggt
     1501 gaccgagacc aagacaagcg tgtccaggca catctacttt ttcagcgcca gaggcaggat
     1561 cgaccccctg gtgtacctgg acgagttcct gaggaacagc gagcccgtgc tgaagagagt
     1621 gaacaggacc ggcaacagca gcagcaacaa gcaggagtac cagctgctga aggacaacct
     1681 ggtgcgcagc tacaacaagg ccctgaagaa gaacgccccc taccccatct tcgctatcaa
     1741 gaacggccct aagagccaca tcggcaggca cctgatgacc agctttctga gcatgaaggg
     1801 cctgaccgag ctgacaaacg tggtgggcaa ctggagcgac aagagggcct ccgccgtggc
     1861 caggaccacc tacacccacc agatcaccgc catccccgac cactacttcg ccctggtgtc
     1921 caggtactac gcctacgacc ccatcagcaa ggagatgatc gccctgaagg acgagaccaa
     1981 ccccatcgag gagtggcagc acatcgagca gctgaagggc agcgccgagg gcagcatcag
     2041 ataccccgcc tggaacggca tcatcagcca ggaggtgctg gactacctga gcagctacat
     2101 caacaggcgg atctgagtcg acggcgcgcc gcggccgcga attcgatatc ataatcaacc
     2161 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     2221 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     2281 cattttctcc tccttgtata aatcctggtt agttcttgcc acggcggaac tcatcgccgc
     2341 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggctcg
     2401 agagatcttc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     2461 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     2521 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     2581 agggggagga ttgggaagac aatagcaggc atgagatctc acgtgcggac cgagcggccg
     2641 caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     2701 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag
     2761 cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc
     2821 ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa
     2881 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc
     2941 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     3001 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     3061 aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     3121 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     3181 cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct
     3241 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     3301 cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     3361 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     3421 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     3481 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     3541 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     3601 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     3661 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     3721 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     3781 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     3841 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     3901 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     3961 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     4021 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     4081 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     4141 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     4201 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     4261 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     4321 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     4381 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     4441 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     4501 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     4561 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     4621 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     4681 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     4741 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     4801 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     4861 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     4921 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     4981 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     5041 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     5101 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     5161 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     5221 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     5281 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     5341 tacggttcct ggccttttgc tggccttttg ctcacatgt
//
