LOCUS       sequence-466374-        5047 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2575m 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 5047)
  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 5047)
  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: AiP20168 - pAAV-AiE2575m-minBG-iCre-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..5047
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      232..241
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             244..264
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(1361..1381)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1568..1679
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1797..1937
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1797..1926
                     /label=AAV2 ITR
     rep_origin      2012..2467
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2099..2118)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2309..2330
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2484..2503)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2603..2625
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2663..2681)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2749..2853
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2854..2922,2923..3714)
                     /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(3072..3091)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3885..4473
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4374..4393
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4535..4664
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 gctatggcac acacagcgag ctgagcgagg aaatcctgcg aagcagggaa actgagagca
       61 taaggcccgg agttgaaaat caagaaggaa agccaagagt ttgagggaaa tgcagaggca
      121 gggttatgtg aggagaacat gccagtgtcc tgtctcttta gccccggagc tcgggctggg
      181 cataaaagtc agggcagagc catctattgc ttacatttgc ttctgggatc cgccaccatg
      241 gtgcccaaga agaagaggaa agtctccaac ctgctgactg tgcaccaaaa cctgcctgcc
      301 ctccctgtgg atgccacctc tgatgaagtc aggaagaacc tgatggacat gttcagggac
      361 aggcaggcct tctctgaaca cacctggaag atgctcctgt ctgtgtgcag atcctgggct
      421 gcctggtgca agctgaacaa caggaaatgg ttccctgctg aacctgagga tgtgagggac
      481 tacctcctgt acctgcaagc cagaggcctg gctgtgaaga ccatccaaca gcacctgggc
      541 cagctcaaca tgctgcacag gagatctggc ctgcctcgcc cttctgactc caatgctgtg
      601 tccctggtga tgaggagaat cagaaaggag aatgtggatg ctggggagag agccaagcag
      661 gccctggcct ttgaacgcac tgactttgac caagtcagat ccctgatgga gaactctgac
      721 agatgccagg acatcaggaa cctggccttc ctgggcattg cctacaacac cctgctgcgc
      781 attgccgaaa ttgccagaat cagagtgaag gacatctccc gcaccgatgg tgggagaatg
      841 ctgatccaca ttggcaggac caagaccctg gtgtccacag ctggtgtgga gaaggccctg
      901 tccctggggg ttaccaagct ggtggagaga tggatctctg tgtctggtgt ggctgatgac
      961 cccaacaact acctgttctg ccgggtcaga aagaatggtg tggctgcccc ttctgccacc
     1021 tcccaactgt ccacccgggc cctggaaggg atctttgagg ccacccaccg cctgatctat
     1081 ggtgccaagg atgactctgg gcagagatac ctggcctggt ctggccactc tgccagagtg
     1141 ggtgctgcca gggacatggc cagggctggt gtgtccatcc ctgaaatcat gcaggctggt
     1201 ggctggacca atgtgaacat tgtgatgaac tacatcagaa acctggactc tgagactggg
     1261 gccatggtga ggctgctcga ggatggggac taagaattcg atatcataat caacctctgg
     1321 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     1381 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     1441 tctcctcctt gtataaatcc tggttagttc ttgccacggc ggaactcatc gccgcctgcc
     1501 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gctcgagaga
     1561 tcttcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     1621 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     1681 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     1741 gaggattggg aagacaatag caggcatgag atctcacgtg cggaccgagc ggccgcagga
     1801 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     1861 gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc
     1921 gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     1981 ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg
     2041 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct
     2101 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     2161 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     2221 cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     2281 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     2341 aactctatct cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg
     2401 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt
     2461 acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     2521 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     2581 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     2641 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     2701 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     2761 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     2821 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     2881 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     2941 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     3001 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     3061 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     3121 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     3181 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     3241 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     3301 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     3361 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     3421 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     3481 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     3541 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     3601 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     3661 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     3721 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     3781 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     3841 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     3901 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     3961 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     4021 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     4081 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     4141 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     4201 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     4261 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     4321 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     4381 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     4441 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     4501 ttcctggcct tttgctggcc ttttgctcac atgtcctgca ggcagctgcg cgctcgctcg
     4561 ctcactgagg ccgcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg
     4621 agcgcgcaga gagggagtgg ccaactccat cactaggggt tcctgcggcc gcattactgc
     4681 agtcacaccg aagggaagat tcaacgagaa tgggaaatta tgagaaatgc ccacacttcc
     4741 tatttcctat atctatttta gttggtggca attacaattt actacgcaac actcaaaata
     4801 taaacggcac tatgtgaaag gatataacag tgacagattt gcagcctcaa agcgtgactt
     4861 caaaatattg gtgccgaaaa ggtttcctgt tccggaaggc agttgagact acaaagccca
     4921 cgcaggtgaa ggcgctggca ggagcgagtg ctctgcgtac cctggatgtt gagcaaagct
     4981 gtgcagggta gatgggaggc gccgcccggc cccatatgga gccgaccttc cgtgccaatc
     5041 gcactat
//
