LOCUS       sequence-471051-        4810 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2569m 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 4810)
  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 4810)
  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: AiP20070 - pAAV-AiE2569m-minBG-iCre(R297T)-BGHpA
FEATURES             Location/Qualifiers
     source          1..4810
                     /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      746..755
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             758..778
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    1844..1955
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2073..2213
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2073..2202
                     /label=AAV2 ITR
     rep_origin      2288..2743
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2375..2394)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2585..2606
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2760..2779)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2879..2901
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2939..2957)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3025..3129
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3130..3198,3199..3990)
                     /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(3348..3367)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4161..4749
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4650..4669
                     /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 gcggccgcag ttattatggc ctgtatgtga gaggcagagg accatgagta
      181 gaaatccatt ctatccattc tctccttcct ccataggaat actagggatc aagctcacat
      241 cactcagtgt cacattgcca gtgctggctc tgttacttca aagcagagga gttcttacta
      301 aatcaagagg attaaaagta taaatacatg tgcacgtagg tgtggaagta tatatatata
      361 tgtacatatt tatgtgtata tacaattata accatgtgtg atggtgactc ttgattgtca
      421 tcataactgg atttagaatc acctaggaga tatgtctctg tgaccagaca acacacactt
      481 ctttctgtcg ccaacgctct aattgagctg ctgtagtcta acttgtcacg acggactaac
      541 tctccaactg caagccagaa gaagtccttc cttccttaag ttgcttttct gcacaaagtg
      601 agaaaaaagt aaccaataag ccaaggtgga actgaagtgc taccaagcag tgctcgatgt
      661 ttcggagctc gggctgggca taaaagtcag ggcagagcca tctattgctt acatttgctt
      721 ctgggatcca gatctttcga agctagccac catggtgccc aagaagaaga ggaaagtctc
      781 caacctgctg actgtgcacc aaaacctgcc tgccctccct gtggatgcca cctctgatga
      841 agtcaggaag aacctgatgg acatgttcag ggacaggcag gccttctctg aacacacctg
      901 gaagatgctc ctgtctgtgt gcagatcctg ggctgcctgg tgcaagctga acaacaggaa
      961 atggttccct gctgaacctg aggatgtgag ggactacctc ctgtacctgc aagccagagg
     1021 cctggctgtg aagaccatcc aacagcacct gggccagctc aacatgctgc acaggagatc
     1081 tggcctgcct cgcccttctg actccaatgc tgtgtccctg gtgatgagga gaatcagaaa
     1141 ggagaatgtg gatgctgggg agagagccaa gcaggccctg gcctttgaac gcactgactt
     1201 tgaccaagtc agatccctga tggagaactc tgacagatgc caggacatca ggaacctggc
     1261 cttcctgggc attgcctaca acaccctgct gcgcattgcc gaaattgcca gaatcagagt
     1321 gaaggacatc tcccgcaccg atggtgggag aatgctgatc cacattggca ggaccaagac
     1381 cctggtgtcc acagctggtg tggagaaggc cctgtccctg ggggttacca agctggtgga
     1441 gagatggatc tctgtgtctg gtgtggctga tgaccccaac aactacctgt tctgccgggt
     1501 cagaaagaat ggtgtggctg ccccttctgc cacctcccaa ctgtccaccc gggccctgga
     1561 agggatcttt gaggccaccc accgcctgat ctatggtgcc aaggatgact ctgggcagag
     1621 atacctggcc tggtctggcc actctgccag agtgggtgct gccaccgaca tggccagggc
     1681 tggtgtgtcc atccctgaaa tcatgcaggc tggtggctgg accaatgtga acattgtgat
     1741 gaactacatc agaaacctgg actctgagac tggggccatg gtgaggctgc tcgaagatgg
     1801 ggactgaggc gcgccgaatt caagcttctc gagagatctt cgactgtgcc ttctagttgc
     1861 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     1921 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     1981 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     2041 catgagatct cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac
     2101 tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc
     2161 gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct
     2221 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa
     2281 ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     2341 gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt
     2401 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     2461 cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt
     2521 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     2581 aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt
     2641 gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa
     2701 aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt
     2761 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     2821 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     2881 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     2941 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     3001 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     3061 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     3121 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     3181 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     3241 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     3301 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     3361 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     3421 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     3481 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     3541 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     3601 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     3661 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     3721 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     3781 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     3841 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     3901 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     3961 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     4021 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     4081 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     4141 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     4201 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     4261 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     4321 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     4381 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     4441 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     4501 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     4561 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     4621 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     4681 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     4741 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     4801 gctcacatgt
//
