LOCUS       sequence-470551-        4646 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0524m 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 4646)
  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 4646)
  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: AiP12873 - pAAV-AiE0524m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4646
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      217..226
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(223..225,226..228,229..939)
                     /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(268..289)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(529..548)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     876..897
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1032..1052)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1239..1350
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1468..1608
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1468..1597
                     /label=AAV2 ITR
     rep_origin      1683..2138
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1770..1789)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1980..2001
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2155..2174)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2274..2296
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2334..2352)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2420..2524
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2525..2593,2594..3385)
                     /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(2743..2762)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3556..4144
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4045..4064
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4206..4335
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 gaagactgca tgcaaatctt aatagaactt tgataattta atgagtgtgg tgaagaaaaa
       61 tggcgatcca gtaccagaac caatactaga acacaatttt ccactctgat gtttaacccc
      121 agggagctcg ggctgggcat aaaagtcagg gcagagccat ctattgctta catttgcttc
      181 tgggatccag atctttcgaa gctagcgcta ccggtcgcca ccatggtgag caagggcgag
      241 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
      301 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
      361 ctgatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgggc
      421 tacggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag
      481 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
      541 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
      601 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
      661 aacagccaca acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc
      721 aagatccgcc acaacatcga ggacggcggc gtgcagctcg ccgaccacta ccagcagaac
      781 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc
      841 aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
      901 gccgccggga tcactctcgg catggacgag ctgtacaagt aagtcgacgg cgcgccgcgg
      961 ccgcgaattc gatatcataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     1021 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     1081 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttagtt
     1141 cttgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     1201 ttgggcactg acaattccgt ggctcgagag atcttcgact gtgccttcta gttgccagcc
     1261 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     1321 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     1381 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatga
     1441 gatctcacgt gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct
     1501 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct
     1561 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc
     1621 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata
     1681 gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     1741 cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     1801 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     1861 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg
     1921 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     1981 tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt
     2041 ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt
     2101 taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat
     2161 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     2221 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     2281 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     2341 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     2401 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     2461 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     2521 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     2581 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     2641 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     2701 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     2761 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     2821 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     2881 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     2941 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     3001 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     3061 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     3121 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     3181 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     3241 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     3301 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     3361 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     3421 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     3481 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     3541 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3601 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3661 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     3721 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3781 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3841 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     3901 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     3961 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     4021 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4081 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4141 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4201 catgtcctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg cgtcgggcga
     4261 cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca
     4321 tcactagggg ttcctgcggc cgcacgcgta gccctgatcc cttggatgaa aatttgtctg
     4381 gtagggttcc cacaggccac ccctcctctc tcaactgata cttagtgctg tgcctcacac
     4441 tcagttgagg ctttgtcact tactggggaa agagatgcca ttagcttcac aattccccaa
     4501 agagacctgc gtggggagtt gtgcgtggca gcctggcaga ggaaacacca ctcgcatcac
     4561 agatatgaaa aactgtaaat ccacaaatga cagattcgtg acttccctgg agaataaata
     4621 gcgagaacag atgggtcctg gggctg
//
