LOCUS       sequence-462680-        4905 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0096h 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 4905)
  AUTHORS   Ben-Simon Y, Hooper M, Narayan S, Daigle TL, Dwivedi D, Way SW, 
            Oster A, Stafford DA, Mich JK, Taormina MJ, Martinez RA, Opitz-Araya
            X, Roth JR, Alexander JR, Allen S, Amster A, Arbuckle J, Ayala A, 
            Baker PM, Bakken TE, Barcelli T, Barta S, Bendrick J, Bertagnolli D,
            Bielstein C, Bishwakarma P, Bowlus J, Boyer G, Brouner K, Casian B, 
            Casper T, Chakka AB, Chakrabarty R, Chance RK, Chavan S, Clark M, 
            Colbert K, Collman F, Daniel S, Departee M, DiValentin P, Donadio N,
            Dotson N, Egdorf T, Fliss T, Gabitto M, Garcia J, Gary A, Gasperini 
            M, Gloe J, Goldy J, Gore BB, Graybuck L, Greisman N, Haeseleer F, 
            Halterman C, Haradon Z, Hastings SD, Helback O, Ho W, Hockemeyer D, 
            Huang C, Huff S, Hunker A, Johansen N, Jones D, Juneau Z, Kalmbach 
            B, Kannan M, Khem S, Kussick E, Kutsal R, Larsen R, Lee C, Lee AY, 
            Leibly M, Lenz GH, Li S, Liang E, Lusk N, Madigan Z, Malloy J, 
            Malone J, McCue R, Melchor J, Mollenkopf T, Moosman S, Morin E, 
            Newman D, Ng L, Ngo K, Omstead V, Otto S, Oyama A, Pena N, Pham T, 
            Phillips E, Pom CA, Potekhina L, Ransford S, Ray PL, Rette D, 
            Reynoldson C, Rimorin C, Rocha D, Ruiz A, Sanchez REA, Sawyer L, 
            Sedeno-Cortes A, Sevigny JP, Shapovalova N, Shepard N, Shulga L, 
            Sigler AR, Siverts L, Soliman S, Somasundaram S, Staats B, Stewart 
            K, Szelenyi E, Tieu M, Trader C, Tran A, van Velthoven CTJ, Walker 
            M, Wang Y, 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 JT, Tasic B
  TITLE     A suite of enhancer AAVs and transgenic mouse lines for genetic 
            access to cortical cell types.
  JOURNAL   Cell. 2025 May 16:S0092-8674(25)00513-6. doi: 
            10.1016/j.cell.2025.05.002.
  PUBMED    40403729
REFERENCE   2  (bases 1 to 4905)
  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: "Cell. 2025 
            May 16:S0092-8674(25)00513-6. doi: 10.1016/j.cell.2025.05.002."
COMMENT     Sequence Label: AiP11542 -
            pAAV-hsA2-AiE0096h-minRho-SYFP2-WPR...
FEATURES             Location/Qualifiers
     source          1..4905
                     /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      942..951
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(948..950,951..953,954..1664)
                     /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(993..1014)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1254..1273)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1601..1622
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1732..1752)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1939..2050
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2168..2308
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2168..2297
                     /label=AAV2 ITR
     rep_origin      2383..2838
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2470..2489)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2680..2701
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2855..2874)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2974..2996
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3034..3052)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3120..3224
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3225..3293,3294..4085)
                     /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(3443..3462)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4256..4844
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4745..4764
                     /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 gcggccgcaa cgcgtttaga acaatggctg gcccatagta aatgccgtgt
      181 tagtgtgtta gttgctgttc ttccacgtca gaagaggcac agacaaatta ccaccaggtg
      241 gcgctcagag tctgcggagg catcacaaca gccctgaatt tgaatcctgc tctgccactg
      301 cctagttgag accttttact acctgactag ctgtttgtgt attttaggtg tttgttttca
      361 gagacccagt gactgatgtt gtcccagaaa gtagaattct tgtgatgcat aaatgatagc
      421 tgatccagcc aacacaactg ttggaggaat tatagcttat agagttgctt ggcagatgaa
      481 atcacttgac taattagaaa ataatgcatt atgctgtgct agaaaaatca cagagcagca
      541 gaagcggaac tgtgttctgg gttgctgtgg gctgccagtc acaattttga gcctgtgcct
      601 tgcattcttt accttttttt ttcccactat ccctacagcc cagaaattga aacagaccct
      661 ggagccttgt gatactgaat atccagcatt cgtctctgag cgcaccatca aggagactac
      721 agggaatatt gcttgtgaag actgctccag aattcgatat cataatcaac cataggtacc
      781 gagctcggga ttcagccggg agcttaggga ggggaggtca cttcataagg gcctgggggg
      841 ggagttggag ccacgagtcg tccagccgga gccccgtgtg gctgagctcc ggcctcagaa
      901 gcatccccgg gttggatcct tcgaagctag cgctaccggt cgccaccatg gtgagcaagg
      961 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     1021 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     1081 tgaagctgat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     1141 tgggctacgg cgtgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct
     1201 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     1261 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     1321 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     1381 actacaacag ccacaacgtc tatatcaccg ccgacaagca gaagaacggc atcaaggcca
     1441 acttcaagat ccgccacaac atcgaggacg gcggcgtgca gctcgccgac cactaccagc
     1501 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagctacc
     1561 agtccaagct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     1621 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaagtc gacatcataa
     1681 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     1741 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     1801 ggctttcatt ttctcctcct tgtataaatc ctggttagtt cttgccacgg cggaactcat
     1861 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     1921 ggctcgagag atcttcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     1981 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     2041 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     2101 acagcaaggg ggaggattgg gaagacaata gcaggcatga gatctcacgt gcggaccgag
     2161 cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc
     2221 actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg
     2281 agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat
     2341 ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg
     2401 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct
     2461 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     2521 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     2581 accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     2641 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     2701 gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt
     2761 cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgggaat tttaacaaaa
     2821 tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     2881 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     2941 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     3001 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     3061 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     3121 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     3181 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     3241 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     3301 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     3361 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     3421 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     3481 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     3541 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     3601 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     3661 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     3721 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     3781 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     3841 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     3901 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     3961 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     4021 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     4081 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     4141 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     4201 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     4261 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     4321 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     4381 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     4441 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     4501 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     4561 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     4621 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     4681 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     4741 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     4801 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     4861 cctttttacg gttcctggcc ttttgctggc cttttgctca catgt
//
