LOCUS       sequence-432584-        5037 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2454m 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 5037)
  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 5037)
  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: AiP1918 - pAAV-AiE2454m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..5037
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(65..84)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     275..296
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(450..469)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     569..591
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(629..647)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        715..819
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(820..888,889..1680)
                     /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(1038..1057)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1851..2439
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2340..2359
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2501..2630
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      3549..3558
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3555..3557,3558..3560,3561..4271)
                     /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(3600..3621)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3861..3880)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4208..4229
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4364..4384)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    4571..4682
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4800..4940
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4800..4929
                     /label=AAV2 ITR
     rep_origin      join(5015..5037,1..433)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg
       61 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
      121 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
      181 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
      241 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
      301 acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc
      361 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
      421 acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
      481 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
      541 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
      601 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
      661 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
      721 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
      781 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
      841 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
      901 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
      961 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     1021 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     1081 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     1141 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     1201 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     1261 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     1321 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     1381 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     1441 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     1501 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     1561 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     1621 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     1681 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     1741 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     1801 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     1861 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     1921 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     1981 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     2041 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     2101 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     2161 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     2221 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     2281 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     2341 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     2401 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     2461 ttacggttcc tggccttttg ctggcctttt gctcacatgt cctgcaggca gctgcgcgct
     2521 cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag
     2581 cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct gcggccgcag
     2641 tatgctttct tatacacacc agctatggct gtcactgacc acattatgct gggccctccc
     2701 atatcaatca ttagtcaaga aaatgctcca cagatttgca tacatgccag tctgatggag
     2761 acattttctc tgttgatcct ccctttcccc agagtatttt agtttgtgtc aatttgatga
     2821 aaatctaacc agtacactgt gctttttatt caaactgtat tccttttcca tgcccagata
     2881 ctaacactga aaaatcctta gtctctgctt cacagggctg agccgacaga gtgaaactcc
     2941 tgaccatcca tccaggaggc agttttcaga gcaagtagca aggctgagtg ctagtggctt
     3001 ttctgttgct gagtcaactt caaaagagga aaggattgct taaagctttc aagccatgat
     3061 ttcttggtcc tggaagcaaa tagcagaagg ggcttgttaa tgttcacttc atattgtaga
     3121 ggaaacaagg agagaagggg gcactatgca ctgttttggg gagatacacc caagtcccta
     3181 accccatcct agcgagccct gcctcttaaa ggatccaaca tttttcaaca gcatacagac
     3241 tggtaagtga gtcttcaata cgcaggactt tgggaacatt tcagattcaa acaataccaa
     3301 ttgagaatga ataaacattt gctttgtttc ttagaggtct gaacagaggc agctgcatca
     3361 aagcgttgac gacatttcaa atcacagtgc aaagcagaca gaaagagctg ctttctgtag
     3421 acaaacatgc ttcagaagct gaacactgga gcccggagct cgggctgggc ataaaagtca
     3481 gggcagagcc atctattgct tacatttgct tctgggatcc agatctttcg aagctagcgc
     3541 taccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     3601 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     3661 gcgatgccac ctacggcaag ctgaccctga agctgatctg caccaccggc aagctgcccg
     3721 tgccctggcc caccctcgtg accaccctgg gctacggcgt gcagtgcttc gcccgctacc
     3781 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     3841 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     3901 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     3961 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcaccgccg
     4021 acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacatc gaggacggcg
     4081 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     4141 tgcccgacaa ccactacctg agctaccagt ccaagctgag caaagacccc aacgagaagc
     4201 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     4261 agctgtacaa gtaagtcgac ggcgcgccgc ggccgcgaat tcgatatcat aatcaacctc
     4321 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     4381 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     4441 ttttctcctc cttgtataaa tcctggttag ttcttgccac ggcggaactc atcgccgcct
     4501 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggctcgag
     4561 agatcttcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     4621 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     4681 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     4741 ggggaggatt gggaagacaa tagcaggcat gagatctcac gtgcggaccg agcggccgca
     4801 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     4861 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg
     4921 agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     4981 tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcatta
//
