LOCUS       sequence-466229-        6537 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2095m 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 6537)
  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 6537)
  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: AiP1264 - pAAV-AiE2095m-minBG-FlpO-WPRE-HGHpA
FEATURES             Location/Qualifiers
     source          1..6537
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      428..437
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(434..439,440..460,461..1732)
                     /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    1757..2345
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1810..1830)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2228..2239)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2377..2853
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2576..2595)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2893..3033
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2893..3022
                     /label=AAV2 ITR
     rep_origin      3108..3563
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3195..3214)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3405..3426
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3580..3599)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3699..3721
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3759..3777)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3845..3949
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3950..4018,4019..4810)
                     /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(4168..4187)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4981..5569
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5470..5489
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   5631..5760
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 ctagccttac aggagaaacg ctgcaacaca agcaacggtg caaacgacat gtgagggact
       61 gagctggtgc tatctacatg gtgcagtgaa gaaggaaata tcttgctggg gcaactgggt
      121 agatgatgtt atgagccaag atggggaatt tggaaagaat ggacttccca gggaaagtcc
      181 tgaaatcttt tttttttgtg ggggggcggg ttgggaatag tgttttttgg attttctgtt
      241 tctattcttg agacaaattc tcactatcta gcccaggctg acctcaaatt catataactc
      301 ttgcctcagc ctccacagta ttgaaattat aaatatgtgt cactgcactc tacacccctt
      361 aaggagctcg ggctgggcat aaaagtcagg gcagagccat ctattgctta catttgcttc
      421 tggcgtggcc accatggctc ctaagaagaa gaggaaggtg atgagccagt tcgacatcct
      481 gtgcaagacc ccccccaagg tgctggtgcg gcagttcgtg gagagattcg agaggcccag
      541 cggcgagaag atcgccagct gtgccgccga gctgacctac ctgtgctgga tgatcaccca
      601 caacggcacc gccatcaaga gggccacctt catgagctac aacaccatca tcagcaacag
      661 cctgagcttc gacatcgtga acaagagcct gcagttcaag tacaagaccc agaaggccac
      721 catcctggag gccagcctga agaagctgat ccccgcctgg gagttcacca tcatccctta
      781 caacggccag aagcaccaga gcgacatcac cgacatcgtg tccagcctgc agctgcagtt
      841 cgagagcagc gaggaggccg acaagggcaa cagccacagc aagaagatgc tgaaggccct
      901 gctgtccgag ggcgagagca tctgggagat caccgagaag atcctgaaca gcttcgagta
      961 caccagcagg ttcaccaaga ccaagaccct gtaccagttc ctgttcctgg ccacattcat
     1021 caactgcggc aggttcagcg acatcaagaa cgtggacccc aagagcttca agctggtgca
     1081 gaacaagtac ctgggcgtga tcattcagtg cctggtgacc gagaccaaga caagcgtgtc
     1141 caggcacatc tactttttca gcgccagagg caggatcgac cccctggtgt acctggacga
     1201 gttcctgagg aacagcgagc ccgtgctgaa gagagtgaac aggaccggca acagcagcag
     1261 caacaagcag gagtaccagc tgctgaagga caacctggtg cgcagctaca acaaggccct
     1321 gaagaagaac gccccctacc ccatcttcgc tatcaagaac ggccctaaga gccacatcgg
     1381 caggcacctg atgaccagct ttctgagcat gaagggcctg accgagctga caaacgtggt
     1441 gggcaactgg agcgacaaga gggcctccgc cgtggccagg accacctaca cccaccagat
     1501 caccgccatc cccgaccact acttcgccct ggtgtccagg tactacgcct acgaccccat
     1561 cagcaaggag atgatcgccc tgaaggacga gaccaacccc atcgaggagt ggcagcacat
     1621 cgagcagctg aagggcagcg ccgagggcag catcagatac cccgcctgga acggcatcat
     1681 cagccaggag gtgctggact acctgagcag ctacatcaac aggcggatct gagaattcga
     1741 tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     1801 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     1861 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     1921 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     1981 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     2041 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     2101 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     2161 ctttccttgg ctgctcgcct atgttgccac ctggattctg cgcgggacgt ccttctgcta
     2221 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     2281 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     2341 cccgcatcga taccgagcgc tgctcgagag atctacgggt ggcatccctg tgacccctcc
     2401 ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct tgtcctaata
     2461 aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat ggggtggagg
     2521 ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg cggggtctat
     2581 tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc cgcctcctgg
     2641 gttcaagcga ttctcctgcc tcagcctccc gagttgttgg gattccaggc atgcatgacc
     2701 aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt ggccaggctg
     2761 gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt gctgggatta
     2821 caggcgtgaa ccactgctcc cttccctgtc cttctgattt tgtaggtaac cacgtgcgga
     2881 ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     2941 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     3001 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta
     3061 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag
     3121 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3181 cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3241 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     3301 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata
     3361 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3421 aactggaaca acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc
     3481 gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     3541 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc
     3601 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     3661 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     3721 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     3781 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     3841 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     3901 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     3961 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     4021 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     4081 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     4141 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     4201 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     4261 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     4321 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     4381 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     4441 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     4501 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     4561 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     4621 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     4681 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     4741 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     4801 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     4861 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     4921 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     4981 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5041 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     5101 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     5161 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     5221 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     5281 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5341 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     5401 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     5461 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     5521 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     5581 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt cctgcaggca
     5641 gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt ggtcgcccgg
     5701 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
     5761 gcggccgcac gcgtctggtt ttctcacaca gctgaacaac tatgccttta agaatcaagc
     5821 ctttgtaact ggaaagatgg ctcaatggtt aaaagaactt gtttctctga aagagaattc
     5881 aggttcaatt catcacccac atggctcaca aacatcctta atttcagttt taatccagac
     5941 aacttctttt gatttccaca agcacaaggc acacagagta tacatgcaga gaaaacattc
     6001 aagtacataa aatagaaata aataaatctt aataactaat caaacattta aaaccataat
     6061 cattcttact ttgaatgttt ttcaagatgt attctactaa atttataaat atttaatgta
     6121 ctttttcatg atagtcatcc ttataggcag tgtcccttgg acaacatatt tgctacagtt
     6181 accacaatga gacaaaaata agacttgttc atttccttcc aataccacaa acatttgctc
     6241 tctcattgcc agacatgatg ctgaatccta agaatagaaa tgaatgaaat atgggtccca
     6301 tgtccagagt tattcattct cgcagaagtc acatgagaac gcaggacatg cacagtgaga
     6361 ggtccaaggc aactggcctt taccagataa gcaagaatca gattataaga ctttggactt
     6421 gacgctgaag tattttgcag agagaggaac ctgaccagac tcaagggatt acccaaagaa
     6481 caatatgagt acacatggca aaggcagact ccgagccaag aagaggagaa gcttagc
//
