LOCUS       sequence-466358-        4703 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE3090m 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 4703)
  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 4703)
  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: AiP20047 - pAAV-AiE3090m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4703
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      246..255
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(252..254,255..257,258..968)
                     /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(297..318)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(558..577)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     905..926
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1061..1081)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1268..1379
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1497..1637
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1497..1626
                     /label=AAV2 ITR
     rep_origin      1712..2167
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1799..1818)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2009..2030
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2184..2203)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2303..2325
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2363..2381)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2449..2553
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2554..2622,2623..3414)
                     /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(2772..2791)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3585..4173
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4074..4093
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4235..4364
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 atgtaattat aattcttact aaggcgctgg cgagtgtcag aggtatttca acaactgccg
       61 agaatacgat aatcaaagcg ccagcattcc tttgttccaa gtcttgttga gggaggctct
      121 caggagatat gctttggagg gaaacgccgg cggagctcgg gctgggcata aaagtcaggg
      181 cagagccatc tattgcttac atttgcttct gggatccaga tctttcgaag ctagcgctac
      241 cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
      301 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
      361 atgccaccta cggcaagctg accctgaagc tgatctgcac caccggcaag ctgcccgtgc
      421 cctggcccac cctcgtgacc accctgggct acggcgtgca gtgcttcgcc cgctaccccg
      481 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
      541 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
      601 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
      661 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc accgccgaca
      721 agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag gacggcggcg
      781 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
      841 ccgacaacca ctacctgagc taccagtcca agctgagcaa agaccccaac gagaagcgcg
      901 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
      961 tgtacaagta agtcgacggc gcgccgcggc cgcgaattcg atatcataat caacctctgg
     1021 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     1081 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     1141 tctcctcctt gtataaatcc tggttagttc ttgccacggc ggaactcatc gccgcctgcc
     1201 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gctcgagaga
     1261 tcttcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     1321 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     1381 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     1441 gaggattggg aagacaatag caggcatgag atctcacgtg cggaccgagc ggccgcagga
     1501 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     1561 gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc
     1621 gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     1681 ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg
     1741 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct
     1801 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     1861 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     1921 cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     1981 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     2041 aactctatct cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg
     2101 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt
     2161 acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     2221 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     2281 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     2341 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     2401 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     2461 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     2521 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     2581 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     2641 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     2701 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     2761 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     2821 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     2881 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     2941 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     3001 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     3061 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     3121 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     3181 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     3241 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     3301 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     3361 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     3421 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     3481 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     3541 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     3601 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     3661 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     3721 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     3781 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     3841 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     3901 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     3961 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     4021 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     4081 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     4141 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     4201 ttcctggcct tttgctggcc ttttgctcac atgtcctgca ggcagctgcg cgctcgctcg
     4261 ctcactgagg ccgcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg
     4321 agcgcgcaga gagggagtgg ccaactccat cactaggggt tcctgcggcc gcacgcgttc
     4381 ctgggggagg gctttccagc caccaccagc ctgttctgcc tggctcttga tctgctcagg
     4441 atcaacggac aacctggctg ggagggagcc cgggcagcct gttctggggg caagtgggca
     4501 cctgtcagac agaggctcca agggaacctg tcagtaccac tcagtttgac ttcctggata
     4561 tagatagccc catcctgaat gactcccacc tacagggctg cccccatctc ccaaggccct
     4621 atttgtgacc cagagggagg gggaaaatcc atcaactctg cagaagcagc taatcagttt
     4681 acttggttgc ctggggtctt cca
//
