LOCUS       sequence-432583-        4622 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2525m 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 4622)
  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 4622)
  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: AiP1941 - pAAV-AiE2525m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4622
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   160..300
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   171..300
                     /label=AAV2 ITR
     polyA_signal    418..529
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     716..736
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(829..1539,1540..1542,1543..1545))
                     /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(871..892)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1220..1239
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1479..1500
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1542..1551
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   2055..2184
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2246..2834)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2326..2345)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3005..3796,3797..3865))
                     /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     3628..3647
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3866..3970)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4038..4056
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4094..4116)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4216..4235
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(join(4252..4622,1..85))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4389..4410)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4601..4620
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ggcgctaagg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
       61 cttaatgcgc cgctacaggg cgcgtactat ggttgctttg acgtatgcgg tgtgaaatac
      121 cgcacagatg cgtaaggaga aaataccgca tcaggcgccc ctgcaggcag ctgcgcgctc
      181 gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt ggtcgcccgg
      241 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
      301 gcggccgctc ggtccgcacg tgagatctca tgcctgctat tgtcttccca atcctccccc
      361 ttgctgtcct gccccacccc accccccaga atagaatgac acctactcag acaatgcgat
      421 gcaatttcct cattttatta ggaaaggaca gtgggagtgg caccttccag ggtcaaggaa
      481 ggcacggggg aggggcaaac aacagatggc tggcaactag aaggcacagt cgaagatctc
      541 tcgagccacg gaattgtcag tgcccaacag ccgagcccct gtccagcagc gggcaaggca
      601 ggcggcgatg agttccgccg tggcaagaac taaccaggat ttatacaagg aggagaaaat
      661 gaaagccata cgggaagcaa tagcatgata caaaggcatt aaagcagcgt atccacatag
      721 cgtaaaagga gcaacatagt taagaatacc agtcaatctt tcacaaattt tgtaatccag
      781 aggttgatta tgatatcgaa ttcgcggccg cggcgcgccg tcgacttact tgtacagctc
      841 gtccatgccg agagtgatcc cggcggcggt cacgaactcc agcaggacca tgtgatcgcg
      901 cttctcgttg gggtctttgc tcagcttgga ctggtagctc aggtagtggt tgtcgggcag
      961 cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga gctgcacgcc
     1021 gccgtcctcg atgttgtggc ggatcttgaa gttggccttg atgccgttct tctgcttgtc
     1081 ggcggtgata tagacgttgt ggctgttgta gttgtactcc agcttgtgcc ccaggatgtt
     1141 gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc
     1201 gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc
     1261 ctggacgtag ccttcgggca tggcggactt gaagaagtcg tgctgcttca tgtggtcggg
     1321 gtagcgggcg aagcactgca cgccgtagcc cagggtggtc acgagggtgg gccagggcac
     1381 gggcagcttg ccggtggtgc agatcagctt cagggtcagc ttgccgtagg tggcatcgcc
     1441 ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag
     1501 gatgggcacc accccggtga acagctcctc gcccttgctc accatggtgg cgaccggtag
     1561 cgctagcttc gaaagatctg gatcccagaa gcaaatgtaa gcaatagatg gctctgccct
     1621 gacttttatg cccagcccga gctccgcaat ttgtcctaag tgtccttcca cccctgggat
     1681 agaggtcatg cctcctcaaa ctgcagttcc ctgagaacta cagagaactc agccctggat
     1741 gggggtgaat gccagcaggt ctcctgctcc cccctagaca ggcagaggcg ttgcttggcc
     1801 tagtgcagcc tgagctcaga acacagtgtt tggtttccag ggctgcaggc ctggtaatgc
     1861 aaatgtcgat tctgtttcca acaaacagtc agcagtccca gtctgcaatc tgcctggagg
     1921 agcccagaaa tcctggtgag acagccatgc ccattgtctc ccaagagtca gctgcagggc
     1981 agggtgccca ccccctttgc ccccagagac acctgtggac ccacgattat tactcttaca
     2041 acgcgtgcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
     2101 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggcggcc tcagtgagcg
     2161 agcgagcgcg cagctgcctg caggacatgt gagcaaaagg ccagcaaaag gccaggaacc
     2221 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     2281 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     2341 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     2401 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     2461 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     2521 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     2581 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     2641 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     2701 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     2761 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     2821 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     2881 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     2941 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     3001 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     3061 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     3121 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     3181 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     3241 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     3301 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     3361 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     3421 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     3481 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     3541 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     3601 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     3661 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     3721 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     3781 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     3841 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     3901 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     3961 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     4021 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     4081 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     4141 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     4201 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat aaaattgtaa
     4261 acgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     4321 aatagaccga aatcggcaaa atcccttata aatcaaaaga atagcccgag atagagttga
     4381 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     4441 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc aaatcaagtt
     4501 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     4561 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     4621 cg
//
