LOCUS       sequence-466330-        4994 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2448m 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 4994)
  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 4994)
  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: AiP1922 - pAAV-AiE2448m-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4994
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     106..127
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(262..282)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    469..580
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   698..838
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   698..827
                     /label=AAV2 ITR
     rep_origin      913..1368
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1000..1019)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1210..1231
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1385..1404)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1504..1526
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1564..1582)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1650..1754
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1755..1823,1824..2615)
                     /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(1973..1992)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2786..3374
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3275..3294
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   3436..3565
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      4441..4450
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4447..4449,4450..4452,4453..4994,1..169)
                     /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(4492..4513)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4753..4772)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
       61 ctaccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
      121 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt aagtcgacgg
      181 cgcgccgcgg ccgcgaattc gatatcataa tcaacctctg gattacaaaa tttgtgaaag
      241 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
      301 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
      361 ctggttagtt cttgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
      421 ggctcggctg ttgggcactg acaattccgt ggctcgagag atcttcgact gtgccttcta
      481 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
      541 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
      601 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
      661 gcaggcatga gatctcacgt gcggaccgag cggccgcagg aacccctagt gatggagttg
      721 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
      781 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg
      841 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa
      901 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
      961 gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt
     1021 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     1081 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt
     1141 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     1201 tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt
     1261 cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt
     1321 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc
     1381 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     1441 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     1501 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     1561 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     1621 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     1681 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     1741 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     1801 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     1861 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     1921 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     1981 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     2041 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     2101 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     2161 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     2221 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     2281 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     2341 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     2401 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     2461 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     2521 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     2581 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     2641 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     2701 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     2761 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     2821 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     2881 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     2941 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     3001 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     3061 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     3121 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     3181 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     3241 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     3301 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     3361 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     3421 cttttgctca catgtcctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg
     3481 cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg
     3541 gccaactcca tcactagggg ttcctgcggc cgcacgcgtt gtaagagtaa taatcgtgtt
     3601 gcttcttcac catgctgtca gagctagtat gtacatgctg ggcaacgggt ggccaagcag
     3661 agtaacttgt cagtgtttaa taaagccctc ctcccttgca gagccaaatg cagctttcct
     3721 cctgagaact ctgcagacga tgacatctcc ctacttggtt tccatagcat ccgttcctga
     3781 agagccctag aaaggccctt ggtcagggcg atccgcccag ctcccctagg aagttggcga
     3841 gcgggttagc caaaagcagc agccaaggtt ccttagcttc ctctgtccgt ggtcctgaaa
     3901 tgctaaactg cgccgcgcgc actgacggga gcggtcaggt tttatggaag gtctcccccg
     3961 gcgctgccta aaccacagct gtgtgtctgt tattcactca cgttcgcaaa gatcagctaa
     4021 cattctcagt cttaattcca ctctgccact ctgccgcaga gcacaattac gccggcgcga
     4081 tgggaggaga ccatgttaag gcaggaagct aacagcagcc gctcacctga ggcctaggaa
     4141 gctcccaagg gttctgagag ctatgagctc tgatcagcaa agtcaccatt ttagacagta
     4201 gggttctttt gcttttttcc ttacacaagg gttcgatgac gtttctggtc aatctgatta
     4261 aaagacagcg gatttgattg cgataagact tcaaaaccgg gaatttacgt tgtttttcgg
     4321 tgaggtgact tccagaacag ggactcatca gcacccgccc aaataccacg gcactgcgcg
     4381 cgccctcggc caccggatcc tccccttcca atgagacttt gtgactgtgc gctaccggtc
     4441 gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     4501 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     4561 acctacggca agctgaccct gaagctgatc tgcaccaccg gcaagctgcc cgtgccctgg
     4621 cccaccctcg tgaccaccct gggctacggc gtgcagtgct tcgcccgcta ccccgaccac
     4681 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     4741 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     4801 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     4861 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcaccgc cgacaagcag
     4921 aagaacggca tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag
     4981 ctcgccgacc acta
//
