LOCUS       sequence-466204-        5924 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2040m 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 5924)
  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 5924)
  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: AiP1176 - pAAV-AiE2040m-minBG-FlpO-WPRE-HGHpA
FEATURES             Location/Qualifiers
     source          1..5924
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      393..402
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(399..404,405..425,426..1697)
                     /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    1722..2310
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1775..1795)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2193..2204)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2342..2818
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2541..2560)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2858..2998
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2858..2987
                     /label=AAV2 ITR
     rep_origin      3073..3528
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3160..3179)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3370..3391
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3545..3564)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3664..3686
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3724..3742)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3810..3914
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3915..3983,3984..4775)
                     /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(4133..4152)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4946..5534
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5435..5454
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   5596..5725
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 aattatcatc agctgtcttg ttgggagtct gaatagagag attaatgagc taataggaaa
       61 ttaagactca gaaagcaatt tccagttatc taaatcgagg cgtttcatct aatcggttca
      121 ggagtgtgga agggagctgc gtgcagagag gccggatgtc agcactattc tgcattgtct
      181 catcagtacg gactcagctg ccagggtctg cccttgcagt gggtttcctg gcttttggtt
      241 gtctgtgtcc caccccactt agagtttctt ccatgggggt ctaaagcccc tagatctcat
      301 ttattggaaa cctttctatc tctgtctcga gctcgggctg ggcataaaag tcagggcaga
      361 gccatctatt gcttacattt gcttctggcg tggccaccat ggctcctaag aagaagagga
      421 aggtgatgag ccagttcgac atcctgtgca agaccccccc caaggtgctg gtgcggcagt
      481 tcgtggagag attcgagagg cccagcggcg agaagatcgc cagctgtgcc gccgagctga
      541 cctacctgtg ctggatgatc acccacaacg gcaccgccat caagagggcc accttcatga
      601 gctacaacac catcatcagc aacagcctga gcttcgacat cgtgaacaag agcctgcagt
      661 tcaagtacaa gacccagaag gccaccatcc tggaggccag cctgaagaag ctgatccccg
      721 cctgggagtt caccatcatc ccttacaacg gccagaagca ccagagcgac atcaccgaca
      781 tcgtgtccag cctgcagctg cagttcgaga gcagcgagga ggccgacaag ggcaacagcc
      841 acagcaagaa gatgctgaag gccctgctgt ccgagggcga gagcatctgg gagatcaccg
      901 agaagatcct gaacagcttc gagtacacca gcaggttcac caagaccaag accctgtacc
      961 agttcctgtt cctggccaca ttcatcaact gcggcaggtt cagcgacatc aagaacgtgg
     1021 accccaagag cttcaagctg gtgcagaaca agtacctggg cgtgatcatt cagtgcctgg
     1081 tgaccgagac caagacaagc gtgtccaggc acatctactt tttcagcgcc agaggcagga
     1141 tcgaccccct ggtgtacctg gacgagttcc tgaggaacag cgagcccgtg ctgaagagag
     1201 tgaacaggac cggcaacagc agcagcaaca agcaggagta ccagctgctg aaggacaacc
     1261 tggtgcgcag ctacaacaag gccctgaaga agaacgcccc ctaccccatc ttcgctatca
     1321 agaacggccc taagagccac atcggcaggc acctgatgac cagctttctg agcatgaagg
     1381 gcctgaccga gctgacaaac gtggtgggca actggagcga caagagggcc tccgccgtgg
     1441 ccaggaccac ctacacccac cagatcaccg ccatccccga ccactacttc gccctggtgt
     1501 ccaggtacta cgcctacgac cccatcagca aggagatgat cgccctgaag gacgagacca
     1561 accccatcga ggagtggcag cacatcgagc agctgaaggg cagcgccgag ggcagcatca
     1621 gataccccgc ctggaacggc atcatcagcc aggaggtgct ggactacctg agcagctaca
     1681 tcaacaggcg gatctgagaa ttcgatatca agcttatcga taatcaacct ctggattaca
     1741 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     1801 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     1861 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     1921 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     1981 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     2041 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     2101 tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctatgtt gccacctgga
     2161 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     2221 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     2281 gtcggatctc cctttgggcc gcctccccgc atcgataccg agcgctgctc gagagatcta
     2341 cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc
     2401 agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc
     2461 cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac
     2521 aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg
     2581 gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt
     2641 gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg
     2701 gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc
     2761 ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct
     2821 gattttgtag gtaaccacgt gcggaccgag cggccgcagg aacccctagt gatggagttg
     2881 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
     2941 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg
     3001 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa
     3061 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     3121 gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt
     3181 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     3241 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt
     3301 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     3361 tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt
     3421 cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt
     3481 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc
     3541 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     3601 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     3661 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     3721 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     3781 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3841 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     3901 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     3961 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     4021 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     4081 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     4141 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     4201 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     4261 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     4321 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     4381 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     4441 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     4501 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     4561 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     4621 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     4681 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     4741 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     4801 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4861 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4921 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4981 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     5041 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     5101 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     5161 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     5221 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     5281 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     5341 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     5401 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     5461 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5521 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     5581 cttttgctca catgtcctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg
     5641 cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg
     5701 gccaactcca tcactagggg ttcctgcggc cgcacgcgtt agaatgcagg ctctttcccc
     5761 ttacctggcc cattaggagc gccaattagt gaaaactggt gccattgctg cctttacaaa
     5821 ttaatgttgg gatagccctc cacccaggaa ggcagatatg tctccttcgt gcctagcccc
     5881 ctcccccctc cacagctctc ctcttaagtg cccacttcct ccgg
//
