LOCUS       sequence-466223-        6538 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2098m 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 6538)
  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 6538)
  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: AiP1247 - pAAV-AiE2098m-minBG-FlpO-WPRE-HGHpA
FEATURES             Location/Qualifiers
     source          1..6538
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   380..509
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(571..1159)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(651..670)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1330..2121,2122..2190))
                     /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     1953..1972
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2191..2295)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2363..2381
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2419..2441)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2541..2560
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2577..3032)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2714..2735)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2926..2945
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3107..3247
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3118..3247
                     /label=AAV2 ITR
     polyA_signal    3287..3763
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     3545..3564
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     misc_feature    3795..4383
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3901..3912
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     4310..4330
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(4408..5679,5680..5700,5701..5706))
                     /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"
     regulatory      5703..5712
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 aacaatagaa tttatttacc aataaagaaa aattaactta ggaaattggc aggaagatgc
       61 ttagaactgg aaaacatact aagtgctgca tcccatgttc ctagaggcaa atatcagata
      121 ttctctgtga caagcaggtt ctagcttcta aatgctaagt ggatgtatct aggtaggtat
      181 aatttgtaga taaaggccag aaaactagaa agaggcccat gagacagaaa aggaaaaaaa
      241 ggaagaaaac ctttaaaaaa caggcacggg aagacaatag aaccatgaga tagatattaa
      301 aacagaagtg aggacactgg aggccgaaga ttatctctgg caaaggggtc aggcaagcag
      361 atcagacgcg tgcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc
      421 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg cggcctcagt
      481 gagcgagcga gcgcgcagct gcctgcagga catgtgagca aaaggccagc aaaaggccag
      541 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
      601 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
      661 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
      721 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
      781 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
      841 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
      901 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
      961 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     1021 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     1081 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     1141 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     1201 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     1261 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     1321 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     1381 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     1441 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     1501 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     1561 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     1621 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     1681 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     1741 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     1801 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     1861 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     1921 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     1981 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     2041 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     2101 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     2161 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     2221 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     2281 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     2341 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     2401 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     2461 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     2521 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accataaaat
     2581 tgtaaacgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     2641 taaccaatag accgaaatcg gcaaaatccc ttataaatca aaagaatagc ccgagataga
     2701 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     2761 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat cacccaaatc
     2821 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     2881 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     2941 aggagcgggc gctaaggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     3001 cgccgcgctt aatgcgccgc tacagggcgc gtactatggt tgctttgacg tatgcggtgt
     3061 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgcccctg caggcagctg
     3121 cgcgctcgct cgctcactga ggccgcccgg gcaaagcccg ggcgtcgggc gacctttggt
     3181 cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg
     3241 ggttcctgcg gccgctcggt ccgcacgtgg ttacctacaa aatcagaagg acagggaagg
     3301 gagcagtggt tcacgcctgt aatcccagca atttgggagg ccaaggtggg tagatcacct
     3361 gagattagga gttggagacc agcctggcca atatggtgaa accccgtctc taccaaaaaa
     3421 acaaaaatta gctgagcctg gtcatgcatg cctggaatcc caacaactcg ggaggctgag
     3481 gcaggagaat cgcttgaacc caggaggcgg agattgcagt gagccaagat tgtgccactg
     3541 cactccagct tggttcccaa tagaccccgc aggccctaca ggttgtcttc ccaacttgcc
     3601 ccttgctcca taccaccccc ctccacccca taatattata gaaggacacc tagtcagaca
     3661 aaatgatgca acttaatttt attaggacaa ggctggtggg cactggagtg gcaacttcca
     3721 gggccaggag aggcactggg gaggggtcac agggatgcca cccgtagatc tctcgagcag
     3781 cgctcggtat cgatgcgggg aggcggccca aagggagatc cgactcgtct gagggcgaag
     3841 gcgaagacgc ggaagaggcc gcagagccgg cagcaggccg cgggaaggaa ggtccgctgg
     3901 attgagggcc gaagggacgt agcagaagga cgtcccgcgc agaatccagg tggcaacata
     3961 ggcgagcagc caaggaaagg acgatgattt ccccgacaac accacggaat tgtcagtgcc
     4021 caacagccga gcccctgtcc agcagcgggc aaggcaggcg gcgatgagtt ccgccgtggc
     4081 aatagggagg gggaaagcga aagtcccgga aaggagctga caggtggtgg caatgcccca
     4141 accagtgggg gttgcgtcag caaacacagt gcacaccacg ccacgttgcc tgacaacggg
     4201 ccacaactcc tcataaagag acagcaacca ggatttatac aaggaggaga aaatgaaagc
     4261 catacgggaa gcaatagcat gatacaaagg cattaaagca gcgtatccac atagcgtaaa
     4321 aggagcaaca tagttaagaa taccagtcaa tctttcacaa attttgtaat ccagaggttg
     4381 attatcgata agcttgatat cgaattctca gatccgcctg ttgatgtagc tgctcaggta
     4441 gtccagcacc tcctggctga tgatgccgtt ccaggcgggg tatctgatgc tgccctcggc
     4501 gctgcccttc agctgctcga tgtgctgcca ctcctcgatg gggttggtct cgtccttcag
     4561 ggcgatcatc tccttgctga tggggtcgta ggcgtagtac ctggacacca gggcgaagta
     4621 gtggtcgggg atggcggtga tctggtgggt gtaggtggtc ctggccacgg cggaggccct
     4681 cttgtcgctc cagttgccca ccacgtttgt cagctcggtc aggcccttca tgctcagaaa
     4741 gctggtcatc aggtgcctgc cgatgtggct cttagggccg ttcttgatag cgaagatggg
     4801 gtagggggcg ttcttcttca gggccttgtt gtagctgcgc accaggttgt ccttcagcag
     4861 ctggtactcc tgcttgttgc tgctgctgtt gccggtcctg ttcactctct tcagcacggg
     4921 ctcgctgttc ctcaggaact cgtccaggta caccaggggg tcgatcctgc ctctggcgct
     4981 gaaaaagtag atgtgcctgg acacgcttgt cttggtctcg gtcaccaggc actgaatgat
     5041 cacgcccagg tacttgttct gcaccagctt gaagctcttg gggtccacgt tcttgatgtc
     5101 gctgaacctg ccgcagttga tgaatgtggc caggaacagg aactggtaca gggtcttggt
     5161 cttggtgaac ctgctggtgt actcgaagct gttcaggatc ttctcggtga tctcccagat
     5221 gctctcgccc tcggacagca gggccttcag catcttcttg ctgtggctgt tgcccttgtc
     5281 ggcctcctcg ctgctctcga actgcagctg caggctggac acgatgtcgg tgatgtcgct
     5341 ctggtgcttc tggccgttgt aagggatgat ggtgaactcc caggcgggga tcagcttctt
     5401 caggctggcc tccaggatgg tggccttctg ggtcttgtac ttgaactgca ggctcttgtt
     5461 cacgatgtcg aagctcaggc tgttgctgat gatggtgttg tagctcatga aggtggccct
     5521 cttgatggcg gtgccgttgt gggtgatcat ccagcacagg taggtcagct cggcggcaca
     5581 gctggcgatc ttctcgccgc tgggcctctc gaatctctcc acgaactgcc gcaccagcac
     5641 cttggggggg gtcttgcaca ggatgtcgaa ctggctcatc accttcctct tcttcttagg
     5701 agccatggtg gccacgccag aagcaaatgt aagcaataga tggctctgcc ctgactttta
     5761 tgcccagccc gagctcgaga tgaagatccc taagaagaga acttgatttc tcctgtatca
     5821 cccgatgatc atgaagggac tgcccaaatt atagcaatgt gtacaaatct agatgttaca
     5881 gatgagaaaa agcatacagt gtttttctga gagtttgttt ggtttgcttg caaggttagt
     5941 tgcatttgtt ttcctgcaca tgacataatt ccatattctt aagaacatga acaataatga
     6001 atgcattctc taaagacagt ataacattga tacaaaaaca ggacaaaaat aggagaaaga
     6061 aaactagatc tattacactg gttatcacag atacaaaact ctaaacaaac taaccatact
     6121 ttgcaaactg aattccaggt cacgttagaa atatctttca ttaattttcc cagaaccagg
     6181 aaaacaaaac aaaacaaaac aaaacaaaac aaaacattcc tcccccggaa gcaagaatgg
     6241 ttactataaa ggaaactggt gacaacagga ggagattgta gggagagagc aatagtgttg
     6301 gtaggagcta aattagtgta gcagttatag atatgagagt ggaagcatct caaaaacaaa
     6361 ctaaaataca actattatat tttctagcta tgtcactcct gggcatatgc ccaaaagact
     6421 ctaaacacta ggagggtaat tgcatatcat atttatagat gctttattca aagagcaagg
     6481 aaatgaaatg gcctagatgt tcattaatag atgaatgggt aacaaacatt caatatgt
//
