LOCUS       sequence-462517-        4935 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2148m 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 4935)
  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 4935)
  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: AiP1377 - pAAV-AiE2148m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4935
                     /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      947..956
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(953..955,956..958,959..1669)
                     /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(998..1019)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1259..1278)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1606..1627
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1762..1782)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1969..2080
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2198..2338
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2198..2327
                     /label=AAV2 ITR
     rep_origin      2413..2868
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2500..2519)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2710..2731
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2885..2904)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3004..3026
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3064..3082)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3150..3254
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3255..3323,3324..4115)
                     /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(3473..3492)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4286..4874
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4775..4794
                     /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 gcgtcaccta agaccagagc cccagggtct ccctagccca
      181 acccaaaaga gaaaggatgc aggaaggatg tggagaggga ggcaggaatc gggccagaag
      241 caccctcaag tcctctcctc ccccctcctc cctcacttcc acttgaggag tcctctcttt
      301 tctctctccc tctcccctcc tccttccttc ccagtcctct ctcccctccc ccttcccttt
      361 tgtcctcccc ccttcagcmc cccctccaat tttctctcct acttctctct ccccttcccc
      421 tcccccctcc cccctcctcc ctgttctttc tccctgccca ggcccactgc ctcccctagc
      481 ccctgctccc agaaagccct ggaaaggcag ctttctttga gacaggaagg tgtcctcaag
      541 tctcaggaag gaaaaacagt cgagctgaac ccaagaccgt ctccaagggg ctggggctgg
      601 ctccgggcca gggctgacat cacctgggaa ccactgcctc tgcagctgca gcccgcctcc
      661 tggggggatg cttctgctat ttaaaaaata ataataatac taaaaaaact tttccagtct
      721 ggtgttttta aatgtgcttt atatagtggg ccctggggtg tgggagattc aatcccctcc
      781 cgtttgtcct cacagcccgg gtttctcaca ccgcactggg cagccttccg aatccacagc
      841 tacttcctgg gaagagctcg ggctgggcat aaaagtcagg gcagagccat ctattgctta
      901 catttgcttc tgggatccag atctttcgaa gctagcgcta ccggtcgcca ccatggtgag
      961 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     1021 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     1081 gaccctgaag ctgatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     1141 caccctgggc tacggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga
     1201 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     1261 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     1321 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     1381 gtacaactac aacagccaca acgtctatat caccgccgac aagcagaaga acggcatcaa
     1441 ggccaacttc aagatccgcc acaacatcga ggacggcggc gtgcagctcg ccgaccacta
     1501 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     1561 ctaccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     1621 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt aagtcgacgg
     1681 cgcgccgcgg ccgcgaattc gatatcataa tcaacctctg gattacaaaa tttgtgaaag
     1741 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     1801 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     1861 ctggttagtt cttgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     1921 ggctcggctg ttgggcactg acaattccgt ggctcgagag atcttcgact gtgccttcta
     1981 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     2041 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     2101 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     2161 gcaggcatga gatctcacgt gcggaccgag cggccgcagg aacccctagt gatggagttg
     2221 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
     2281 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg
     2341 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa
     2401 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     2461 gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt
     2521 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     2581 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt
     2641 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     2701 tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt
     2761 cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt
     2821 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc
     2881 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     2941 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     3001 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     3061 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     3121 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3181 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     3241 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     3301 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     3361 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     3421 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     3481 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     3541 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     3601 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     3661 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     3721 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     3781 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     3841 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     3901 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     3961 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     4021 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     4081 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     4141 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4201 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4261 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4321 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4381 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     4441 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4501 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4561 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4621 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4681 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4741 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     4801 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     4861 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     4921 cttttgctca catgt
//
