LOCUS       sequence-470464-        4701 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0482m 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 4701)
  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 4701)
  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: AiP12322 - pAAV-AiE0482m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4701
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   213..342
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(404..992)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(484..503)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1163..1954,1955..2023))
                     /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     1786..1805
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2024..2128)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2196..2214
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2252..2274)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2374..2393
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2410..2865)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2547..2568)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2759..2778
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2940..3080
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2951..3080
                     /label=AAV2 ITR
     polyA_signal    3198..3309
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3496..3516
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3609..4319,4320..4322,4323..4325))
                     /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(3651..3672)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4000..4019
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4259..4280
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4322..4331
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 aaagctgccc tcgccccagt cctcagcact gtgaatagat tgcttcaagt ggcaaaaggg
       61 actttgcaga tgtgactgag ggtctagact ctgaggtgga gagattatct gtgtggtccc
      121 gatctgatca catggaccct tgggagtgaa gaattattct tactataatt aaaacaagga
      181 taagagagag aagactgtac gcgtgcggcc gcaggaaccc ctagtgatgg agttggccac
      241 tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc
      301 gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggacatgtga gcaaaaggcc
      361 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
      421 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
      481 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
      541 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
      601 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
      661 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
      721 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
      781 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
      841 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
      901 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
      961 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     1021 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     1081 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     1141 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     1201 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     1261 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     1321 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     1381 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     1441 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     1501 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     1561 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     1621 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     1681 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     1741 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     1801 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     1861 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     1921 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     1981 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     2041 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     2101 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     2161 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     2221 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     2281 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     2341 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     2401 tgcaccataa aattgtaaac gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     2461 tcagctcatt ttttaaccaa tagaccgaaa tcggcaaaat cccttataaa tcaaaagaat
     2521 agcccgagat agagttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     2581 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     2641 catcacccaa atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     2701 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     2761 ggaagaaagc gaaaggagcg ggcgctaagg cgctggcaag tgtagcggtc acgctgcgcg
     2821 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtactat ggttgctttg
     2881 acgtatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgccc
     2941 ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg
     3001 ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa
     3061 ctccatcact aggggttcct gcggccgctc ggtccgcacg tgagatctca tgcctgctat
     3121 tgtcttccca atcctccccc ttgctgtcct gccccacccc accccccaga atagaatgac
     3181 acctactcag acaatgcgat gcaatttcct cattttatta ggaaaggaca gtgggagtgg
     3241 caccttccag ggtcaaggaa ggcacggggg aggggcaaac aacagatggc tggcaactag
     3301 aaggcacagt cgaagatctc tcgagccacg gaattgtcag tgcccaacag ccgagcccct
     3361 gtccagcagc gggcaaggca ggcggcgatg agttccgccg tggcaagaac taaccaggat
     3421 ttatacaagg aggagaaaat gaaagccata cgggaagcaa tagcatgata caaaggcatt
     3481 aaagcagcgt atccacatag cgtaaaagga gcaacatagt taagaatacc agtcaatctt
     3541 tcacaaattt tgtaatccag aggttgatta tgatatcgaa ttcgcggccg cggcgcgccg
     3601 tcgacttact tgtacagctc gtccatgccg agagtgatcc cggcggcggt cacgaactcc
     3661 agcaggacca tgtgatcgcg cttctcgttg gggtctttgc tcagcttgga ctggtagctc
     3721 aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag
     3781 tggtcggcga gctgcacgcc gccgtcctcg atgttgtggc ggatcttgaa gttggccttg
     3841 atgccgttct tctgcttgtc ggcggtgata tagacgttgt ggctgttgta gttgtactcc
     3901 agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg
     3961 ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc
     4021 ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca tggcggactt gaagaagtcg
     4081 tgctgcttca tgtggtcggg gtagcgggcg aagcactgca cgccgtagcc cagggtggtc
     4141 acgagggtgg gccagggcac gggcagcttg ccggtggtgc agatcagctt cagggtcagc
     4201 ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg
     4261 tcgccgtcca gctcgaccag gatgggcacc accccggtga acagctcctc gcccttgctc
     4321 accatggtgg cgaccggtag cgctagcttc gaaagatctg gatcccagaa gcaaatgtaa
     4381 gcaatagatg gctctgccct gacttttatg cccagcccga gctcatcagg gcctggagtt
     4441 tgggtcacag catctaaaag gcagagattc cattacctgg ggtggtagga gatgtgagag
     4501 aagcagattt ggggaaagac gttcaagagc tcagccttgg gcaaacgcat gttggagggg
     4561 tctgttaaat gaccacatgg accacatgga gatgtgcatg atggaatgcc cagttctgca
     4621 tttcagagga gctgacaggg aggaattagc tatttagaca cagtaatgat tgtgactgtg
     4681 ataggcatac taatgttctc c
//
