LOCUS       sequence-460661-        5085 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0459m_3xC2 is an enhancer sequence, designed to drive 
            AAV-mediated transgene expression in specific brain cell types.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5085)
  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 5085)
  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: AiP14187 -
            pAAV-AiE0459m_3xC2-minBG-iCre(R297T...
FEATURES             Location/Qualifiers
     source          1..5085
                     /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      1021..1030
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1033..1053
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2119..2230
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2348..2488
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2348..2477
                     /label=AAV2 ITR
     rep_origin      2563..3018
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2650..2669)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2860..2881
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3035..3054)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3154..3176
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3214..3232)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3300..3404
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3405..3473,3474..4265)
                     /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(3623..3642)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4436..5024
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4925..4944
                     /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 gcgtctatga atagttacaa gtgagaaaga gaaatggaga
      181 ttttaccatt taatcatctc cgaagagaag gtgagaagaa attgagaaag gtaatttgca
      241 taccatgcat aggcattaga gaaagcattt tttttcttaa ctctccattt gtttacagat
      301 atttccatat ggccttttaa aaacttcaac agagaaattc gaacacaact aactggaatt
      361 tcaattttca tataaaacca gcatcataga tatatatgta gctactctcc tatgaatagt
      421 tacaagtgag aaagagaaat ggagatttta ccatttaatc atctccgaag agaaggtgag
      481 aagaaattga gaaaggtaat ttgcatacca tgcataggca ttagagaaag catttttttt
      541 cttaactctc catttgttta cagatatttc catatggcct tttaaaaact tcaacagaga
      601 aattcgaaca caactaactg gaatttcaat tttcatataa aaccagcatc atagatatat
      661 atgtagctac tctcctatga atagttacaa gtgagaaaga gaaatggaga ttttaccatt
      721 taatcatctc cgaagagaag gtgagaagaa attgagaaag gtaatttgca taccatgcat
      781 aggcattaga gaaagcattt tttttcttaa ctctccattt gtttacagat atttccatat
      841 ggccttttaa aaacttcaac agagaaattc gaacacaact aactggaatt tcaattttca
      901 tataaaacca gcatcataga tatatatgta gctactctcg agctcgggct gggcataaaa
      961 gtcagggcag agccatctat tgcttacatt tgcttctggg atccagatct ttcgaagcta
     1021 gccaccatgg tgcccaagaa gaagaggaaa gtctccaacc tgctgactgt gcaccaaaac
     1081 ctgcctgccc tccctgtgga tgccacctct gatgaagtca ggaagaacct gatggacatg
     1141 ttcagggaca ggcaggcctt ctctgaacac acctggaaga tgctcctgtc tgtgtgcaga
     1201 tcctgggctg cctggtgcaa gctgaacaac aggaaatggt tccctgctga acctgaggat
     1261 gtgagggact acctcctgta cctgcaagcc agaggcctgg ctgtgaagac catccaacag
     1321 cacctgggcc agctcaacat gctgcacagg agatctggcc tgcctcgccc ttctgactcc
     1381 aatgctgtgt ccctggtgat gaggagaatc agaaaggaga atgtggatgc tggggagaga
     1441 gccaagcagg ccctggcctt tgaacgcact gactttgacc aagtcagatc cctgatggag
     1501 aactctgaca gatgccagga catcaggaac ctggccttcc tgggcattgc ctacaacacc
     1561 ctgctgcgca ttgccgaaat tgccagaatc agagtgaagg acatctcccg caccgatggt
     1621 gggagaatgc tgatccacat tggcaggacc aagaccctgg tgtccacagc tggtgtggag
     1681 aaggccctgt ccctgggggt taccaagctg gtggagagat ggatctctgt gtctggtgtg
     1741 gctgatgacc ccaacaacta cctgttctgc cgggtcagaa agaatggtgt ggctgcccct
     1801 tctgccacct cccaactgtc cacccgggcc ctggaaggga tctttgaggc cacccaccgc
     1861 ctgatctatg gtgccaagga tgactctggg cagagatacc tggcctggtc tggccactct
     1921 gccagagtgg gtgctgccac cgacatggcc agggctggtg tgtccatccc tgaaatcatg
     1981 caggctggtg gctggaccaa tgtgaacatt gtgatgaact acatcagaaa cctggactct
     2041 gagactgggg ccatggtgag gctgctcgaa gatggggact gaggcgcgcc gaattcaagc
     2101 ttctcgagag atcttcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     2161 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     2221 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     2281 acagcaaggg ggaggattgg gaagacaata gcaggcatga gatctcacgt gcggaccgag
     2341 cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc
     2401 actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg
     2461 agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat
     2521 ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg
     2581 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct
     2641 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     2701 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     2761 accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     2821 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     2881 gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt
     2941 cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     3001 tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     3061 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     3121 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     3181 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     3241 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     3301 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     3361 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     3421 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     3481 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     3541 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     3601 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     3661 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     3721 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     3781 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     3841 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     3901 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     3961 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     4021 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     4081 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     4141 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     4201 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     4261 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     4321 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     4381 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     4441 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     4501 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     4561 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     4621 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     4681 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     4741 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     4801 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     4861 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     4921 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     4981 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     5041 cctttttacg gttcctggcc ttttgctggc cttttgctca catgt
//
