LOCUS       sequence-466192-        5839 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0128h is an enhancer sequence, designed to drive AAV-mediated 
            transgene expression in specific populations of brain cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5839)
  AUTHORS   Ben-Simon Y, Hooper M, Narayan S, Daigle T, Dwivedi D, Way SW, Oster
            A, Stafford DA, Mich JK, Taormina MJ, Martinez RA, Opitz-Araya X, 
            Roth JR, Allen S, Ayala A, Bakken TE, Barcelli T, Barta S, Bendrick 
            J, Bertagnolli D, Bowlus J, Boyer G, Brouner K, Casian B, Casper T, 
            Chakka AB, Chakrabarty R, Chance RK, Chavan S, Departee M, Donadio 
            N, Dotson N, Egdorf T, Gabitto M, Garcia J, Gary A, Gasperini M, 
            Goldy J, Gore BB, Graybuck L, Greisman N, Haeseleer F, Halterman C, 
            Helback O, Hockemeyer D, Huang C, Huff S, Hunker A, Johansen N, 
            Juneau Z, Kalmbach B, Khem S, Kussick E, Kutsal R, Larsen R, Lee C, 
            Lee AY, Leibly M, Lenz GH, Liang E, Lusk N, Malone J, Mollenkopf T, 
            Morin E, Newman D, Ng L, Ngo K, Omstead V, Oyama A, Pham T, Pom CA, 
            Potekhina L, Ransford S, Rette D, Rimorin C, Rocha D, Ruiz A, 
            Sanchez REA, Sedeno-Cortes A, Sevigny JP, Shapovalova N, Shulga L, 
            Sigler AR, Siverts LA, Somasundaram S, Stewart K, Szelenyi E, Tieu 
            M, Trader C, van Velthoven CTJ, Walker M, Weed N, Wirthlin M, Wood 
            T, Wynalda B, Yao Z, Zhou T, Ariza J, Dee N, Reding M, Ronellenfitch
            K, Mufti S, Sunkin SM, Smith KA, Esposito L, Waters J, Thyagarajan 
            B, Yao S, Lein ES, Zeng H, Levi BP, Ngai J, Ting J, Tasic B
  TITLE     A suite of enhancer AAVs and transgenic mouse lines for genetic 
            access to cortical cell types (preprint).
  JOURNAL   bioRxiv [Preprint]. 2024 Sep 26:2024.06.10.597244. doi: 
            10.1101/2024.06.10.597244.
  PUBMED    38915722
REFERENCE   2  (bases 1 to 5839)
  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"; journalName: "bioRxiv 
            [Preprint]. 2024 Sep 26:2024.06.10.597244. doi: 
            10.1101/2024.06.10.597244."
COMMENT     Sequence Label: AiP1233 - pAAV-AiE0128h-minBG-FlpO-WPRE-HGHpA
FEATURES             Location/Qualifiers
     source          1..5839
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   337..466
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(528..1116)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(608..627)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1287..2078,2079..2147))
                     /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     1910..1929
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2148..2252)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2320..2338
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2376..2398)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2498..2517
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2534..2989)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2671..2692)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2883..2902
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3064..3204
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3075..3204
                     /label=AAV2 ITR
     polyA_signal    3244..3720
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     3502..3521
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     misc_feature    3752..4340
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3858..3869
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     4267..4287
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(4365..5636,5637..5657,5658..5663))
                     /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"
     regulatory      5660..5669
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 ctttctgagc cttcacgcta tcagaagagg ctgtcaggtg gagcactgag ctgctggatg
       61 cagctggagg aggctggagg actgcttgtt caaaaatttg ttgaaaatat tcaggcagtt
      121 ggtggcagct ggatttgatg accttgtggt tattttcaac tcaacgattc atgtatcctc
      181 tgagggaggc tccattcctg ttactaattt tgtcatgttt cccatgggga ttctgttgca
      241 aagtatttat cccctgatct cctctcaggg gtccctcagc tggcttccct tccctgcttt
      301 gagatcctcc tcatgatact agacgcgtgc ggccgcagga acccctagtg atggagttgg
      361 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
      421 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggacat gtgagcaaaa
      481 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
      541 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
      601 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
      661 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
      721 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
      781 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
      841 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
      901 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
      961 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     1021 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     1081 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     1141 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     1201 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     1261 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     1321 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     1381 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     1441 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     1501 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     1561 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     1621 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     1681 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     1741 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     1801 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     1861 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     1921 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     1981 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     2041 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     2101 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     2161 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     2221 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     2281 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     2341 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     2401 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     2461 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     2521 agagtgcacc ataaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt
     2581 taaatcagct cattttttaa ccaatagacc gaaatcggca aaatccctta taaatcaaaa
     2641 gaatagcccg agatagagtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     2701 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     2761 gaaccatcac ccaaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     2821 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     2881 gaagggaaga aagcgaaagg agcgggcgct aaggcgctgg caagtgtagc ggtcacgctg
     2941 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgta ctatggttgc
     3001 tttgacgtat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
     3061 gcccctgcag gcagctgcgc gctcgctcgc tcactgaggc cgcccgggca aagcccgggc
     3121 gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg
     3181 ccaactccat cactaggggt tcctgcggcc gctcggtccg cacgtggtta cctacaaaat
     3241 cagaaggaca gggaagggag cagtggttca cgcctgtaat cccagcaatt tgggaggcca
     3301 aggtgggtag atcacctgag attaggagtt ggagaccagc ctggccaata tggtgaaacc
     3361 ccgtctctac caaaaaaaca aaaattagct gagcctggtc atgcatgcct ggaatcccaa
     3421 caactcggga ggctgaggca ggagaatcgc ttgaacccag gaggcggaga ttgcagtgag
     3481 ccaagattgt gccactgcac tccagcttgg ttcccaatag accccgcagg ccctacaggt
     3541 tgtcttccca acttgcccct tgctccatac cacccccctc caccccataa tattatagaa
     3601 ggacacctag tcagacaaaa tgatgcaact taattttatt aggacaaggc tggtgggcac
     3661 tggagtggca acttccaggg ccaggagagg cactggggag gggtcacagg gatgccaccc
     3721 gtagatctct cgagcagcgc tcggtatcga tgcggggagg cggcccaaag ggagatccga
     3781 ctcgtctgag ggcgaaggcg aagacgcgga agaggccgca gagccggcag caggccgcgg
     3841 gaaggaaggt ccgctggatt gagggccgaa gggacgtagc agaaggacgt cccgcgcaga
     3901 atccaggtgg caacataggc gagcagccaa ggaaaggacg atgatttccc cgacaacacc
     3961 acggaattgt cagtgcccaa cagccgagcc cctgtccagc agcgggcaag gcaggcggcg
     4021 atgagttccg ccgtggcaat agggaggggg aaagcgaaag tcccggaaag gagctgacag
     4081 gtggtggcaa tgccccaacc agtgggggtt gcgtcagcaa acacagtgca caccacgcca
     4141 cgttgcctga caacgggcca caactcctca taaagagaca gcaaccagga tttatacaag
     4201 gaggagaaaa tgaaagccat acgggaagca atagcatgat acaaaggcat taaagcagcg
     4261 tatccacata gcgtaaaagg agcaacatag ttaagaatac cagtcaatct ttcacaaatt
     4321 ttgtaatcca gaggttgatt atcgataagc ttgatatcga attctcagat ccgcctgttg
     4381 atgtagctgc tcaggtagtc cagcacctcc tggctgatga tgccgttcca ggcggggtat
     4441 ctgatgctgc cctcggcgct gcccttcagc tgctcgatgt gctgccactc ctcgatgggg
     4501 ttggtctcgt ccttcagggc gatcatctcc ttgctgatgg ggtcgtaggc gtagtacctg
     4561 gacaccaggg cgaagtagtg gtcggggatg gcggtgatct ggtgggtgta ggtggtcctg
     4621 gccacggcgg aggccctctt gtcgctccag ttgcccacca cgtttgtcag ctcggtcagg
     4681 cccttcatgc tcagaaagct ggtcatcagg tgcctgccga tgtggctctt agggccgttc
     4741 ttgatagcga agatggggta gggggcgttc ttcttcaggg ccttgttgta gctgcgcacc
     4801 aggttgtcct tcagcagctg gtactcctgc ttgttgctgc tgctgttgcc ggtcctgttc
     4861 actctcttca gcacgggctc gctgttcctc aggaactcgt ccaggtacac cagggggtcg
     4921 atcctgcctc tggcgctgaa aaagtagatg tgcctggaca cgcttgtctt ggtctcggtc
     4981 accaggcact gaatgatcac gcccaggtac ttgttctgca ccagcttgaa gctcttgggg
     5041 tccacgttct tgatgtcgct gaacctgccg cagttgatga atgtggccag gaacaggaac
     5101 tggtacaggg tcttggtctt ggtgaacctg ctggtgtact cgaagctgtt caggatcttc
     5161 tcggtgatct cccagatgct ctcgccctcg gacagcaggg ccttcagcat cttcttgctg
     5221 tggctgttgc ccttgtcggc ctcctcgctg ctctcgaact gcagctgcag gctggacacg
     5281 atgtcggtga tgtcgctctg gtgcttctgg ccgttgtaag ggatgatggt gaactcccag
     5341 gcggggatca gcttcttcag gctggcctcc aggatggtgg ccttctgggt cttgtacttg
     5401 aactgcaggc tcttgttcac gatgtcgaag ctcaggctgt tgctgatgat ggtgttgtag
     5461 ctcatgaagg tggccctctt gatggcggtg ccgttgtggg tgatcatcca gcacaggtag
     5521 gtcagctcgg cggcacagct ggcgatcttc tcgccgctgg gcctctcgaa tctctccacg
     5581 aactgccgca ccagcacctt ggggggggtc ttgcacagga tgtcgaactg gctcatcacc
     5641 ttcctcttct tcttaggagc catggtggcc acgccagaag caaatgtaag caatagatgg
     5701 ctctgccctg acttttatgc ccagcccgag ctcctataaa tataccacct ggatggagga
     5761 gtgtgttaga tccagacccc agtatgctgc cctctgccca ctagaggcct gccccaggat
     5821 gtgctgggcc ccacagggg
//
