LOCUS       sequence-466347-        4816 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE3047m 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 4816)
  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 4816)
  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: AiP20015 - pAAV-AiE3047m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4816
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      302..311
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(308..310,311..313,314..1024)
                     /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(353..374)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(614..633)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     961..982
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1117..1137)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1324..1435
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1553..1693
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1553..1682
                     /label=AAV2 ITR
     rep_origin      1768..2223
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1855..1874)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2065..2086
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2240..2259)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2359..2381
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2419..2437)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2505..2609
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2610..2678,2679..3470)
                     /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(2828..2847)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3641..4229
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4130..4149
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4291..4420
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 tcgttaaaat gactagtaag tctcaaagac caagtagctg cctggggttt cattccattt
       61 ctgtgaggcc catatgcatc tcaaaacacc ctgttctcat taatctgtct tttgccagtt
      121 caattagcag ggtcctcagc cactgcaact gagaaggtgg aagagaaagc ttttgttctt
      181 ctttggcaga ccattactta cgccggcgga gctcgggctg ggcataaaag tcagggcaga
      241 gccatctatt gcttacattt gcttctggga tccagatctt tcgaagctag cgctaccggt
      301 cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
      361 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
      421 cacctacggc aagctgaccc tgaagctgat ctgcaccacc ggcaagctgc ccgtgccctg
      481 gcccaccctc gtgaccaccc tgggctacgg cgtgcagtgc ttcgcccgct accccgacca
      541 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
      601 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
      661 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
      721 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcaccg ccgacaagca
      781 gaagaacggc atcaaggcca acttcaagat ccgccacaac atcgaggacg gcggcgtgca
      841 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
      901 caaccactac ctgagctacc agtccaagct gagcaaagac cccaacgaga agcgcgatca
      961 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     1021 caagtaagtc gacggcgcgc cgcggccgcg aattcgatat cataatcaac ctctggatta
     1081 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     1141 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     1201 ctccttgtat aaatcctggt tagttcttgc cacggcggaa ctcatcgccg cctgccttgc
     1261 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggctc gagagatctt
     1321 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     1381 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     1441 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     1501 attgggaaga caatagcagg catgagatct cacgtgcgga ccgagcggcc gcaggaaccc
     1561 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
     1621 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc
     1681 agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     1741 caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg
     1801 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt
     1861 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     1921 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     1981 atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     2041 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact
     2101 ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa
     2161 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
     2221 ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     2281 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     2341 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     2401 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     2461 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     2521 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     2581 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     2641 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     2701 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     2761 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     2821 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     2881 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     2941 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     3001 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     3061 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     3121 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     3181 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     3241 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     3301 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     3361 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     3421 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     3481 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     3541 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     3601 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     3661 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     3721 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     3781 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     3841 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     3901 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     3961 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     4021 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     4081 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     4141 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     4201 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     4261 tggccttttg ctggcctttt gctcacatgt cctgcaggca gctgcgcgct cgctcgctca
     4321 ctgaggccgc ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg
     4381 cgcagagagg gagtggccaa ctccatcact aggggttcct gcggccgcac gcgttgagcc
     4441 atgcaaatga gctgaaggca tcagaatgtg ctaccactca gccaccttcg cataaaaact
     4501 gctttgagct ggaagcaacc aacaccaaca aaagctccct atgctgccct tgtttgccaa
     4561 aaagtagagc ctgatttcac tttgtgaggt gttgctccct ctgcctctct cgtgccaggg
     4621 aaggcagaat gaaaagccac tctaggctgt ttgtccagag gcggcactgc aaggcacagc
     4681 gtcactaaca tggctgaaac agtccttctc ctccagtacc ttcccccatg tattcagctt
     4741 cccaccagcc actgcccttg gacgccagga aaccttttcc tctgccttgt catataccca
     4801 taaaattgtt cccccc
//
