LOCUS       sequence-466208-        6191 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0466m 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 6191)
  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 6191)
  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: AiP1190 - pAAV-AiE0466m-minBG-FlpO-WPRE-HGHpA
FEATURES             Location/Qualifiers
     source          1..6191
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      258..267
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(264..269,270..290,291..1562)
                     /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    1587..2175
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1640..1660)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2058..2069)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2207..2683
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2406..2425)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2723..2863
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2723..2852
                     /label=AAV2 ITR
     rep_origin      2938..3393
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3025..3044)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3235..3256
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3410..3429)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3529..3551
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3589..3607)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3675..3779
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3780..3848,3849..4640)
                     /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(3998..4017)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4811..5399
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5300..5319
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   5461..5590
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 ctattcccgt ctgctcccag ggctcttcaa agctcatgag agtgcttcct gtgattgatc
       61 tggaaaacca agcaccttca aagtgggaaa cattattgca attactttta gattgggaga
      121 ttattatcaa tcattttcat ttgattactt gatgaacaac ctaacacaaa atcggaggcc
      181 attctacctt aaggagctcg ggctgggcat aaaagtcagg gcagagccat ctattgctta
      241 catttgcttc tggcgtggcc accatggctc ctaagaagaa gaggaaggtg atgagccagt
      301 tcgacatcct gtgcaagacc ccccccaagg tgctggtgcg gcagttcgtg gagagattcg
      361 agaggcccag cggcgagaag atcgccagct gtgccgccga gctgacctac ctgtgctgga
      421 tgatcaccca caacggcacc gccatcaaga gggccacctt catgagctac aacaccatca
      481 tcagcaacag cctgagcttc gacatcgtga acaagagcct gcagttcaag tacaagaccc
      541 agaaggccac catcctggag gccagcctga agaagctgat ccccgcctgg gagttcacca
      601 tcatccctta caacggccag aagcaccaga gcgacatcac cgacatcgtg tccagcctgc
      661 agctgcagtt cgagagcagc gaggaggccg acaagggcaa cagccacagc aagaagatgc
      721 tgaaggccct gctgtccgag ggcgagagca tctgggagat caccgagaag atcctgaaca
      781 gcttcgagta caccagcagg ttcaccaaga ccaagaccct gtaccagttc ctgttcctgg
      841 ccacattcat caactgcggc aggttcagcg acatcaagaa cgtggacccc aagagcttca
      901 agctggtgca gaacaagtac ctgggcgtga tcattcagtg cctggtgacc gagaccaaga
      961 caagcgtgtc caggcacatc tactttttca gcgccagagg caggatcgac cccctggtgt
     1021 acctggacga gttcctgagg aacagcgagc ccgtgctgaa gagagtgaac aggaccggca
     1081 acagcagcag caacaagcag gagtaccagc tgctgaagga caacctggtg cgcagctaca
     1141 acaaggccct gaagaagaac gccccctacc ccatcttcgc tatcaagaac ggccctaaga
     1201 gccacatcgg caggcacctg atgaccagct ttctgagcat gaagggcctg accgagctga
     1261 caaacgtggt gggcaactgg agcgacaaga gggcctccgc cgtggccagg accacctaca
     1321 cccaccagat caccgccatc cccgaccact acttcgccct ggtgtccagg tactacgcct
     1381 acgaccccat cagcaaggag atgatcgccc tgaaggacga gaccaacccc atcgaggagt
     1441 ggcagcacat cgagcagctg aagggcagcg ccgagggcag catcagatac cccgcctgga
     1501 acggcatcat cagccaggag gtgctggact acctgagcag ctacatcaac aggcggatct
     1561 gagaattcga tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat
     1621 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     1681 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     1741 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     1801 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     1861 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     1921 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     1981 aatcatcgtc ctttccttgg ctgctcgcct atgttgccac ctggattctg cgcgggacgt
     2041 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     2101 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     2161 gggccgcctc cccgcatcga taccgagcgc tgctcgagag atctacgggt ggcatccctg
     2221 tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct
     2281 tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat
     2341 ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg
     2401 cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc
     2461 cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg gattccaggc
     2521 atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt
     2581 ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt
     2641 gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt tgtaggtaac
     2701 cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg
     2761 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     2821 cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat
     2881 tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg
     2941 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     3001 cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     3061 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     3121 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
     3181 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     3241 tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt gatttataag
     3301 ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     3361 cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct
     3421 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     3481 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     3541 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     3601 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     3661 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     3721 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     3781 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     3841 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     3901 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     3961 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     4021 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     4081 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     4141 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     4201 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     4261 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     4321 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     4381 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     4441 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     4501 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     4561 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     4621 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     4681 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     4741 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     4801 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     4861 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     4921 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     4981 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     5041 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     5101 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     5161 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     5221 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     5281 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     5341 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     5401 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     5461 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
     5521 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
     5581 aggggttcct gcggccgcac gcgtgtccta tatgtcacct gccctacctc cagagctccg
     5641 atggcttcgg ggactgttgg gatattagtt tagctttttt attgggttct cttacctcca
     5701 actcttattc ctccccaatc ccttccaagc ctccaacttt agataggaga gagagaaggt
     5761 tagaggggaa agaggcatag agctccttcg cctgctgatt aggggcatcc acttccttgg
     5821 ggcaagtcta ctctttatcg tcaggatatc ttcaacttct catcaagcca caacaacagc
     5881 aatgatgagc agcagcctcc acaggccctc ttggggaatc tcccctttat accctctcca
     5941 gagtccccag aattaaacta tctgcagctg gcaaaaatca cacctctgct aggtcgtgaa
     6001 acaaatcata atcacctgcc gtgaagcagc ctcttgtccc acgcctggga ttccaacaaa
     6061 aacgcattca catagcataa ctaggctttt aaagaaacca agattctaac tactacaggg
     6121 gatgcttcag taaatacgca ctgggcactg acaagcaaga aatgctgagc acctgacatg
     6181 aagtcacgtt a
//
