LOCUS       sequence-466337-        4977 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2453m 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 4977)
  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 4977)
  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: AiP2025 - pAAV-AiE2453m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4977
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   223..352
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(414..1002)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(494..513)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1173..1964,1965..2033))
                     /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     1796..1815
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2034..2138)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2206..2224
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2262..2284)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2384..2403
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2420..2875)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2557..2578)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2769..2788
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2950..3090
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2961..3090
                     /label=AAV2 ITR
     polyA_signal    3208..3319
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3506..3526
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3619..4329,4330..4332,4333..4335))
                     /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(3661..3682)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4010..4029
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4269..4290
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4332..4341
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 tatgcttttc tatcctagga ctaaaacctg aaaagagagg tgtatattag aggaaaaggc
       61 aaaactacta tatgaagcca tggggaatgc actaaaaata cccctcacca ataattcaca
      121 aagagaatta gttgccagat ggtttcagtg ccattccaaa atgtatatac tgaactccaa
      181 gcttcagtat gatcacctta ggagggggac gcgtgcggcc gcaggaaccc ctagtgatgg
      241 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
      301 cccgacgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggacatgtga
      361 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
      421 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
      481 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
      541 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
      601 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
      661 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
      721 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
      781 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
      841 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
      901 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
      961 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     1021 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     1081 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     1141 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     1201 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     1261 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     1321 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     1381 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     1441 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     1501 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     1561 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     1621 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     1681 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     1741 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     1801 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     1861 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     1921 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     1981 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     2041 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     2101 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     2161 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     2221 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     2281 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     2341 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     2401 gtactgagag tgcaccataa aattgtaaac gttaatattt tgttaaaatt cgcgttaaat
     2461 ttttgttaaa tcagctcatt ttttaaccaa tagaccgaaa tcggcaaaat cccttataaa
     2521 tcaaaagaat agcccgagat agagttgagt gttgttccag tttggaacaa gagtccacta
     2581 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     2641 ctacgtgaac catcacccaa atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     2701 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     2761 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaagg cgctggcaag tgtagcggtc
     2821 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtactat
     2881 ggttgctttg acgtatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     2941 tcaggcgccc ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc
     3001 ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg
     3061 gagtggccaa ctccatcact aggggttcct gcggccgctc ggtccgcacg tgagatctca
     3121 tgcctgctat tgtcttccca atcctccccc ttgctgtcct gccccacccc accccccaga
     3181 atagaatgac acctactcag acaatgcgat gcaatttcct cattttatta ggaaaggaca
     3241 gtgggagtgg caccttccag ggtcaaggaa ggcacggggg aggggcaaac aacagatggc
     3301 tggcaactag aaggcacagt cgaagatctc tcgagccacg gaattgtcag tgcccaacag
     3361 ccgagcccct gtccagcagc gggcaaggca ggcggcgatg agttccgccg tggcaagaac
     3421 taaccaggat ttatacaagg aggagaaaat gaaagccata cgggaagcaa tagcatgata
     3481 caaaggcatt aaagcagcgt atccacatag cgtaaaagga gcaacatagt taagaatacc
     3541 agtcaatctt tcacaaattt tgtaatccag aggttgatta tgatatcgaa ttcgcggccg
     3601 cggcgcgccg tcgacttact tgtacagctc gtccatgccg agagtgatcc cggcggcggt
     3661 cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttgc tcagcttgga
     3721 ctggtagctc aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt
     3781 ctgctggtag tggtcggcga gctgcacgcc gccgtcctcg atgttgtggc ggatcttgaa
     3841 gttggccttg atgccgttct tctgcttgtc ggcggtgata tagacgttgt ggctgttgta
     3901 gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag
     3961 ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt
     4021 gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca tggcggactt
     4081 gaagaagtcg tgctgcttca tgtggtcggg gtagcgggcg aagcactgca cgccgtagcc
     4141 cagggtggtc acgagggtgg gccagggcac gggcagcttg ccggtggtgc agatcagctt
     4201 cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg
     4261 gccgtttacg tcgccgtcca gctcgaccag gatgggcacc accccggtga acagctcctc
     4321 gcccttgctc accatggtgg cgaccggtag cgctagcttc gaaagatctg gatcccagaa
     4381 gcaaatgtaa gcaatagatg gctctgccct gacttttatg cccagcccga gctccggcaa
     4441 atatggttta tgaaaaaact tcctcatatt ctgcagcaat cattggtaat tttctaacat
     4501 aatttaaaat cttaatgaga aatgatttaa agcagaaatt tcttttgatt ctgatcatta
     4561 cctatgcaac ctgttaagga tcatatgcat gatattagtt cttaatggtt tgtttctaga
     4621 tgggcctcgt actgaagttt cattctttta ttaaatctag gaagctggtc catgtgaaag
     4681 gggctattgt tccctgataa ttacccagct gctctcttca tggcttgcct ctctgccatg
     4741 ttcagcaatt taaagttttt tttttcttta aataggaaca gtaaataaat gcaggtggga
     4801 aatttctaaa tataaaattg ttttgtttgt aacttagaaa cataatttgt atacaacctt
     4861 agtacttcta tccagcattg atgaaaatgt ggccaggcat ttcaatcttt gcttttgcaa
     4921 gaaccgaaga gactgttctt ctctttttat agtaacaaca gataatttaa gagttaa
//
