LOCUS       sequence-461958-        5914 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2052m 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 5914)
  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 5914)
  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: AiP1212 - pAAV-AiE2052m-minBG-FlpO-WPRE-HGHpA
FEATURES             Location/Qualifiers
     source          1..5914
                     /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      712..721
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(718..723,724..744,745..2016)
                     /codon_start=1
                     /product="nuclear-targeted site-specific recombinase"
                     /label=FLPo
                     /note="FLPo is a mouse codon-optimized fusion between the 
                     SV40 nuclear localization signal and a thermostable version
                     of the Saccharomyces cerevisiae site-specific recombinase 
                     FLP.  Recombination occurs at FRT sequences."
                     /translation="MAPKKKRKVMSQFDILCKTPPKVLVRQFVERFERPSGEKIASCAA
                     ELTYLCWMITHNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKK
                     LIPAWEFTIIPYNGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESI
                     WEITEKILNSFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGV
                     IIQCLVTETKTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEY
                     QLLKDNLVRSYNKALKKNAPYPIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWS
                     DKRASAVARTTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQL
                     KGSAEGSIRYPAWNGIISQEVLDYLSSYINRRI"
     misc_feature    2041..2629
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2094..2114)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2512..2523)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2661..3137
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2860..2879)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3177..3317
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3177..3306
                     /label=AAV2 ITR
     rep_origin      3392..3847
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3479..3498)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3689..3710
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3864..3883)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3983..4005
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4043..4061)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4129..4233
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4234..4302,4303..5094)
                     /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(4452..4471)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5265..5853
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5754..5773
                     /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 gcggccgcac gcgtatatgg atgtcttacc tgcatgaata cttgtgtacc
      181 acatgtatgc ctttctccat ggaggccaga agagggtatc agattccctg gaactgaagt
      241 tacagatagt tgtgagctgc catgtgggta ctggaaatac atgtgtctaa ttttaaggac
      301 cagctcctac tgaggacata aatgttatgt ttcttaaaag tctataccat aagaatcttc
      361 ttcagatctt ggtgtgaaga ttcttacccc agattggtta cagaagcggt accaggtggg
      421 tggatcccat caaaagcttt agtgagaaat cacccttttc cttcacagca gcccagctga
      481 ctggtgaact ttgccgaact gtaaaatatc aaacctttgg gaagctttct gcaacacgag
      541 aagttttaat ccttcaatat taaaaagaaa aataagaaaa aaaaactgtg gagaagtcac
      601 gaacaggaga aacaaaaagg aattgtgcac agggtcagtc gcttaaggag ctcgggctgg
      661 gcataaaagt cagggcagag ccatctattg cttacatttg cttctggcgt ggccaccatg
      721 gctcctaaga agaagaggaa ggtgatgagc cagttcgaca tcctgtgcaa gacccccccc
      781 aaggtgctgg tgcggcagtt cgtggagaga ttcgagaggc ccagcggcga gaagatcgcc
      841 agctgtgccg ccgagctgac ctacctgtgc tggatgatca cccacaacgg caccgccatc
      901 aagagggcca ccttcatgag ctacaacacc atcatcagca acagcctgag cttcgacatc
      961 gtgaacaaga gcctgcagtt caagtacaag acccagaagg ccaccatcct ggaggccagc
     1021 ctgaagaagc tgatccccgc ctgggagttc accatcatcc cttacaacgg ccagaagcac
     1081 cagagcgaca tcaccgacat cgtgtccagc ctgcagctgc agttcgagag cagcgaggag
     1141 gccgacaagg gcaacagcca cagcaagaag atgctgaagg ccctgctgtc cgagggcgag
     1201 agcatctggg agatcaccga gaagatcctg aacagcttcg agtacaccag caggttcacc
     1261 aagaccaaga ccctgtacca gttcctgttc ctggccacat tcatcaactg cggcaggttc
     1321 agcgacatca agaacgtgga ccccaagagc ttcaagctgg tgcagaacaa gtacctgggc
     1381 gtgatcattc agtgcctggt gaccgagacc aagacaagcg tgtccaggca catctacttt
     1441 ttcagcgcca gaggcaggat cgaccccctg gtgtacctgg acgagttcct gaggaacagc
     1501 gagcccgtgc tgaagagagt gaacaggacc ggcaacagca gcagcaacaa gcaggagtac
     1561 cagctgctga aggacaacct ggtgcgcagc tacaacaagg ccctgaagaa gaacgccccc
     1621 taccccatct tcgctatcaa gaacggccct aagagccaca tcggcaggca cctgatgacc
     1681 agctttctga gcatgaaggg cctgaccgag ctgacaaacg tggtgggcaa ctggagcgac
     1741 aagagggcct ccgccgtggc caggaccacc tacacccacc agatcaccgc catccccgac
     1801 cactacttcg ccctggtgtc caggtactac gcctacgacc ccatcagcaa ggagatgatc
     1861 gccctgaagg acgagaccaa ccccatcgag gagtggcagc acatcgagca gctgaagggc
     1921 agcgccgagg gcagcatcag ataccccgcc tggaacggca tcatcagcca ggaggtgctg
     1981 gactacctga gcagctacat caacaggcgg atctgagaat tcgatatcaa gcttatcgat
     2041 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     2101 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     2161 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     2221 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     2281 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     2341 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     2401 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
     2461 gcctatgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     2521 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     2581 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccga
     2641 gcgctgctcg agagatctac gggtggcatc cctgtgaccc ctccccagtg cctctcctgg
     2701 ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta agttgcatca
     2761 ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg gtatggagca
     2821 aggggcaagt tgggaagaca acctgtaggg cctgcggggt ctattgggaa ccaagctgga
     2881 gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc
     2941 tgcctcagcc tcccgagttg ttgggattcc aggcatgcat gaccaggctc agctaatttt
     3001 tgtttttttg gtagagacgg ggtttcacca tattggccag gctggtctcc aactcctaat
     3061 ctcaggtgat ctacccacct tggcctccca aattgctggg attacaggcg tgaaccactg
     3121 ctcccttccc tgtccttctg attttgtagg taaccacgtg cggaccgagc ggccgcagga
     3181 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     3241 gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc
     3301 gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     3361 ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg
     3421 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct
     3481 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3541 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     3601 cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     3661 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     3721 aactctatct cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg
     3781 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt
     3841 acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     3901 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     3961 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     4021 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     4081 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     4141 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4201 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     4261 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     4321 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     4381 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     4441 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     4501 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     4561 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     4621 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     4681 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     4741 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     4801 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     4861 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     4921 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     4981 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     5041 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     5101 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5161 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     5221 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     5281 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     5341 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     5401 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     5461 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     5521 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     5581 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     5641 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     5701 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     5761 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     5821 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     5881 ttcctggcct tttgctggcc ttttgctcac atgt
//
