LOCUS       sequence-461971-        4815 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2547m 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 4815)
  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 4815)
  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: AiP1954 - pAAV-AiE2547m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4815
                     /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      827..836
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(833..835,836..838,839..1549)
                     /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(878..899)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1139..1158)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1486..1507
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1642..1662)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1849..1960
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2078..2218
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2078..2207
                     /label=AAV2 ITR
     rep_origin      2293..2748
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2380..2399)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2590..2611
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2765..2784)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2884..2906
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2944..2962)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3030..3134
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3135..3203,3204..3995)
                     /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(3353..3372)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4166..4754
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4655..4674
                     /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 gcgttgtaag agtaataatc gccaaaacat taaaactaga
      181 ttgacaaatc acactgtctc aagccacctt tccagagtcc actgccagac cccggagcct
      241 gcaggctact tttctctgac atcacctggc tgccattgtt ttgctgataa aggacactcc
      301 agggagactg aaaagctaga agagtgagaa gggcttctgt ttcctgtttg cttgctgtgg
      361 gtgttagcat gactcagtaa caatgctgca tgccaatctc tcatttcctg caccaccccc
      421 aaccccgaac tgccctcggg ttctcagtgc agaggttcaa cttggttctt ttgagctcat
      481 cttcctcact tccagggcag cagcagctcc actaaagcag gttccctccc ggactctcag
      541 ctccttaggg ccctttctct gagctgcagg tatcaggatc aggtgagagg cacccagcac
      601 tcagaggtgg agcccctccc ctttgcttcc tggcccccag ggaaggagtt atttcagcag
      661 ctacattctc tgtactttct ttatcctttc ttttttagtt ctccaaatcc tgttttccca
      721 gctcctatat ccggagctcg ggctgggcat aaaagtcagg gcagagccat ctattgctta
      781 catttgcttc tgggatccag atctttcgaa gctagcgcta ccggtcgcca ccatggtgag
      841 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
      901 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
      961 gaccctgaag ctgatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     1021 caccctgggc tacggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga
     1081 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     1141 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     1201 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     1261 gtacaactac aacagccaca acgtctatat caccgccgac aagcagaaga acggcatcaa
     1321 ggccaacttc aagatccgcc acaacatcga ggacggcggc gtgcagctcg ccgaccacta
     1381 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     1441 ctaccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     1501 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt aagtcgacgg
     1561 cgcgccgcgg ccgcgaattc gatatcataa tcaacctctg gattacaaaa tttgtgaaag
     1621 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     1681 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     1741 ctggttagtt cttgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     1801 ggctcggctg ttgggcactg acaattccgt ggctcgagag atcttcgact gtgccttcta
     1861 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     1921 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     1981 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     2041 gcaggcatga gatctcacgt gcggaccgag cggccgcagg aacccctagt gatggagttg
     2101 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
     2161 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg
     2221 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa
     2281 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     2341 gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt
     2401 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     2461 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt
     2521 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     2581 tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt
     2641 cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt
     2701 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc
     2761 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     2821 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     2881 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     2941 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     3001 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3061 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     3121 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     3181 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     3241 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     3301 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     3361 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     3421 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     3481 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     3541 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     3601 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     3661 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     3721 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     3781 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     3841 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     3901 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     3961 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     4021 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4081 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4141 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4201 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4261 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     4321 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4381 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4441 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4501 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4561 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4621 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     4681 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     4741 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     4801 cttttgctca catgt
//
