LOCUS       sequence-450576-        5020 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0456m is an enhancer sequence, designed to drive AAV-mediated 
            transgene expression in cortical L5_ET neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5020)
  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 5020)
  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: AiP12828 - pAAV-AiE0456m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..5020
                     /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      1032..1041
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1038..1040,1041..1043,1044..1754)
                     /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(1083..1104)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1344..1363)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1691..1712
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1847..1867)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2054..2165
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2283..2423
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2283..2412
                     /label=AAV2 ITR
     rep_origin      2498..2953
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2585..2604)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2795..2816
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2970..2989)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3089..3111
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3149..3167)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3235..3339
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3340..3408,3409..4200)
                     /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(3558..3577)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4371..4959
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4860..4879
                     /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 gcgttggtta gcactatttt gaatggacac ttctataaaa
      181 tttgttttaa aaacaatatg gaagagtcat aagatgtctg ggcacagtgc cccagtcaac
      241 tcatcaatgg gattctggaa tccaaaggtt atatggtgta aaaacacaga gagcaaaatg
      301 cacaaggatc ttcatagaac cataattttt aaaaagcaaa cacttccccc aaaggagcat
      361 gtaagacaac ttgaaggtat taaaatggga ttgtggtttt attagttaga gaacatcatg
      421 caaattatat tatctaggca ttataatgca aactttaaaa ggaaatgctc attatgtaat
      481 gttaagtgaa aaaaatcagg ccataaaatg aaatgtgcat tatgcatgca tatgatgaaa
      541 agacaagaaa tagataagaa tatgcagaaa tacaaatagt tggctgtggt gtgggcttct
      601 ggcttgtttt ctgttttaaa gccattacat catctttcca atgacaaact acattttaag
      661 ttattgaaaa caaaatctcg ttagccttgt ccttgcataa attcccagag aacacattat
      721 tcttgttccc ttctacctct gctacttgtc tttgatttgg acctgctcca cctcctctta
      781 cttttcagtc ctgaactaga tggaaaaaag aaatagaccc tcaggaaaca aggaagcaaa
      841 accacccaac tgtgtggaag ccgtgcattc aagctgcaaa cgagtggagt ggcttcaata
      901 ccttaactta tccccattgt tgcaaaggga gacccagaga gctcgggctg ggcataaaag
      961 tcagggcaga gccatctatt gcttacattt gcttctggga tccagatctt tcgaagctag
     1021 cgctaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     1081 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     1141 agggcgatgc cacctacggc aagctgaccc tgaagctgat ctgcaccacc ggcaagctgc
     1201 ccgtgccctg gcccaccctc gtgaccaccc tgggctacgg cgtgcagtgc ttcgcccgct
     1261 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     1321 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     1381 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     1441 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcaccg
     1501 ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac atcgaggacg
     1561 gcggcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     1621 tgctgcccga caaccactac ctgagctacc agtccaagct gagcaaagac cccaacgaga
     1681 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     1741 acgagctgta caagtaagtc gacggcgcgc cgcggccgcg aattcgatat cataatcaac
     1801 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     1861 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     1921 tcattttctc ctccttgtat aaatcctggt tagttcttgc cacggcggaa ctcatcgccg
     1981 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggctc
     2041 gagagatctt cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     2101 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     2161 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     2221 aagggggagg attgggaaga caatagcagg catgagatct cacgtgcgga ccgagcggcc
     2281 gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
     2341 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga
     2401 gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg
     2461 cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta
     2521 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg
     2581 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     2641 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     2701 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     2761 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     2821 acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc
     2881 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     2941 acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     3001 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     3061 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     3121 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     3181 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     3241 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     3301 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     3361 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     3421 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     3481 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     3541 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     3601 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     3661 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     3721 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     3781 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     3841 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     3901 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     3961 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     4021 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     4081 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     4141 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     4201 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     4261 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     4321 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     4381 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     4441 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     4501 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     4561 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     4621 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     4681 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     4741 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     4801 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     4861 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     4921 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     4981 ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
