LOCUS       sequence-459485-        5613 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0140h_3xC2 is an enhancer sequence, designed to drive 
            AAV-mediated transgene expression in cortical and striatal Pvalb 
            interneurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5613)
  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 5613)
  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: AiP15100:
            pAAV-AiE0140h_3xC2-minBG-FlpO-4X2C-W...
FEATURES             Location/Qualifiers
     source          1..5613
                     /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      826..835
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(832..837,838..858,859..2130)
                     /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(2440..2460)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2647..2758
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2876..3016
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2876..3005
                     /label=AAV2 ITR
     rep_origin      3091..3546
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3178..3197)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3388..3409
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3563..3582)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3682..3704
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3742..3760)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3828..3932
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3933..4001,4002..4793)
                     /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(4151..4170)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4964..5552
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5453..5472
                     /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 agctagccac catggctcct
      841 aagaagaaga ggaaggtgat gagccagttc gacatcctgt gcaagacccc ccccaaggtg
      901 ctggtgcggc agttcgtgga gagattcgag aggcccagcg gcgagaagat cgccagctgt
      961 gccgccgagc tgacctacct gtgctggatg atcacccaca acggcaccgc catcaagagg
     1021 gccaccttca tgagctacaa caccatcatc agcaacagcc tgagcttcga catcgtgaac
     1081 aagagcctgc agttcaagta caagacccag aaggccacca tcctggaggc cagcctgaag
     1141 aagctgatcc ccgcctggga gttcaccatc atcccttaca acggccagaa gcaccagagc
     1201 gacatcaccg acatcgtgtc cagcctgcag ctgcagttcg agagcagcga ggaggccgac
     1261 aagggcaaca gccacagcaa gaagatgctg aaggccctgc tgtccgaggg cgagagcatc
     1321 tgggagatca ccgagaagat cctgaacagc ttcgagtaca ccagcaggtt caccaagacc
     1381 aagaccctgt accagttcct gttcctggcc acattcatca actgcggcag gttcagcgac
     1441 atcaagaacg tggaccccaa gagcttcaag ctggtgcaga acaagtacct gggcgtgatc
     1501 attcagtgcc tggtgaccga gaccaagaca agcgtgtcca ggcacatcta ctttttcagc
     1561 gccagaggca ggatcgaccc cctggtgtac ctggacgagt tcctgaggaa cagcgagccc
     1621 gtgctgaaga gagtgaacag gaccggcaac agcagcagca acaagcagga gtaccagctg
     1681 ctgaaggaca acctggtgcg cagctacaac aaggccctga agaagaacgc cccctacccc
     1741 atcttcgcta tcaagaacgg ccctaagagc cacatcggca ggcacctgat gaccagcttt
     1801 ctgagcatga agggcctgac cgagctgaca aacgtggtgg gcaactggag cgacaagagg
     1861 gcctccgccg tggccaggac cacctacacc caccagatca ccgccatccc cgaccactac
     1921 ttcgccctgg tgtccaggta ctacgcctac gaccccatca gcaaggagat gatcgccctg
     1981 aaggacgaga ccaaccccat cgaggagtgg cagcacatcg agcagctgaa gggcagcgcc
     2041 gagggcagca tcagataccc cgcctggaac ggcatcatca gccaggaggt gctggactac
     2101 ctgagcagct acatcaacag gcggatctga gtcgacggcg cgccgcggcc ttaaagagac
     2161 cggttcactg tgacagtaaa agagaccggt tcactgtgag aatgaaagag accggttcac
     2221 tgtgatcgga aaagagaccg gttcactgtg agcggccttg aaacccagca gacaatgtag
     2281 ctcagtagaa acccagcaga caatgtagct gaatggaaac ccagcagaca atgtagcttc
     2341 ggagaaaccc agcagacaat gtagctgtcg acgaattcga tatcataatc aacctctgga
     2401 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     2461 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     2521 ctcctccttg tataaatcct ggttagttct tgccacggcg gaactcatcg ccgcctgcct
     2581 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg ctcgagagat
     2641 cttcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     2701 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     2761 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     2821 aggattggga agacaatagc aggcatgaga tctcacgtgc ggaccgagcg gccgcaggaa
     2881 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg
     2941 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg
     3001 cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     3061 tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg
     3121 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc
     3181 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3241 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     3301 ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     3361 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3421 actctatctc gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt
     3481 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     3541 caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     3601 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     3661 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     3721 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     3781 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     3841 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     3901 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     3961 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     4021 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     4081 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     4141 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     4201 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     4261 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     4321 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     4381 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     4441 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     4501 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     4561 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     4621 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     4681 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     4741 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     4801 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     4861 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     4921 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     4981 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     5041 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     5101 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     5161 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     5221 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     5281 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     5341 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     5401 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     5461 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     5521 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     5581 tcctggcctt ttgctggcct tttgctcaca tgt
//
