LOCUS       sequence-432597-        4888 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2475m 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 4888)
  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 4888)
  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: AiP1965 - pAAV-AiE2475m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4888
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   503..632
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(694..1282)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(774..793)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1453..2244,2245..2313))
                     /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     2076..2095
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2314..2418)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2486..2504
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2542..2564)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2664..2683
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2700..3155)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2837..2858)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3049..3068
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3230..3370
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3241..3370
                     /label=AAV2 ITR
     polyA_signal    3488..3599
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3786..3806
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3899..4609,4610..4612,4613..4615))
                     /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(3941..3962)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4290..4309
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4549..4570
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4612..4621
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caaggcatgt agccatctct tccctctttc tcttcttcct tctagatctt gtcttcctgc
       61 gccctcattg ctcgcgtttc tgattcatgt atctgaccac catcagcaga tctgaagagg
      121 acaggaagca cccagttcga actcttccaa gtcggggcct gcacacccca cccccacccc
      181 cttcctggag tctgtctggg aggggatccc caatgacgca ctcgggaagg cagtacagag
      241 ccccacgcca ggcccgggga tcatgaatga gggcagagag aaggccaggc gcttcctcct
      301 caccagccgg agctttggcc cccagcaggg tgagcagctg ccctcctgag gcagcccagg
      361 caggctctat gcccagccct tgttggccag aagacaagct ggctgggtcc ccgcgaaaga
      421 ggtcgtttca atagctttca ccctgcactc ccgcacacca ccccagcccc ggccccgcat
      481 gctgtaacct ccttgcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg
      541 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggcggcctc
      601 agtgagcgag cgagcgcgca gctgcctgca ggacatgtga gcaaaaggcc agcaaaaggc
      661 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
      721 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
      781 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
      841 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
      901 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
      961 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     1021 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     1081 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     1141 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     1201 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     1261 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     1321 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     1381 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     1441 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     1501 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     1561 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     1621 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     1681 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     1741 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     1801 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     1861 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     1921 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     1981 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     2041 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     2101 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     2161 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     2221 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     2281 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     2341 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     2401 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     2461 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     2521 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     2581 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     2641 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccataa
     2701 aattgtaaac gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     2761 ttttaaccaa tagaccgaaa tcggcaaaat cccttataaa tcaaaagaat agcccgagat
     2821 agagttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     2881 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccaa
     2941 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     3001 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     3061 gaaaggagcg ggcgctaagg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     3121 acccgccgcg cttaatgcgc cgctacaggg cgcgtactat ggttgctttg acgtatgcgg
     3181 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgccc ctgcaggcag
     3241 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
     3301 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
     3361 aggggttcct gcggccgctc ggtccgcacg tgagatctca tgcctgctat tgtcttccca
     3421 atcctccccc ttgctgtcct gccccacccc accccccaga atagaatgac acctactcag
     3481 acaatgcgat gcaatttcct cattttatta ggaaaggaca gtgggagtgg caccttccag
     3541 ggtcaaggaa ggcacggggg aggggcaaac aacagatggc tggcaactag aaggcacagt
     3601 cgaagatctc tcgagccacg gaattgtcag tgcccaacag ccgagcccct gtccagcagc
     3661 gggcaaggca ggcggcgatg agttccgccg tggcaagaac taaccaggat ttatacaagg
     3721 aggagaaaat gaaagccata cgggaagcaa tagcatgata caaaggcatt aaagcagcgt
     3781 atccacatag cgtaaaagga gcaacatagt taagaatacc agtcaatctt tcacaaattt
     3841 tgtaatccag aggttgatta tgatatcgaa ttcgcggccg cggcgcgccg tcgacttact
     3901 tgtacagctc gtccatgccg agagtgatcc cggcggcggt cacgaactcc agcaggacca
     3961 tgtgatcgcg cttctcgttg gggtctttgc tcagcttgga ctggtagctc aggtagtggt
     4021 tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga
     4081 gctgcacgcc gccgtcctcg atgttgtggc ggatcttgaa gttggccttg atgccgttct
     4141 tctgcttgtc ggcggtgata tagacgttgt ggctgttgta gttgtactcc agcttgtgcc
     4201 ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg
     4261 tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga
     4321 tggtgcgctc ctggacgtag ccttcgggca tggcggactt gaagaagtcg tgctgcttca
     4381 tgtggtcggg gtagcgggcg aagcactgca cgccgtagcc cagggtggtc acgagggtgg
     4441 gccagggcac gggcagcttg ccggtggtgc agatcagctt cagggtcagc ttgccgtagg
     4501 tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca
     4561 gctcgaccag gatgggcacc accccggtga acagctcctc gcccttgctc accatggtgg
     4621 cgaccggtag cgctagcttc gaaagatctg gatcccagaa gcaaatgtaa gcaatagatg
     4681 gctctgccct gacttttatg cccagcccga gctccggcta cgtgacagtt ccctagagaa
     4741 tggtacagct gcaagcctca acacccagca tgcaccgcag ctggggagtg ctgggagcct
     4801 ttgaagagcc cgggcaggat gccacaggca tctgcgggaa gcaggggata cacaaggcag
     4861 ggtcagcact gactgatttc aaccacta
//
