LOCUS       sequence-466214-        5856 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2057m 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 5856)
  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 5856)
  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: AiP1218 - pAAV-AiE2057m-minBG-FlpO-WPRE-HGHpA
FEATURES             Location/Qualifiers
     source          1..5856
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   183..312
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(374..962)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(454..473)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1133..1924,1925..1993))
                     /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     1756..1775
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1994..2098)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2166..2184
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2222..2244)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2344..2363
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2380..2835)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2517..2538)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2729..2748
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2910..3050
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2921..3050
                     /label=AAV2 ITR
     polyA_signal    3090..3566
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     3348..3367
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     misc_feature    3598..4186
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3704..3715
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     4113..4133
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(4211..5482,5483..5503,5504..5509))
                     /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      5506..5515
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 agagaagtgg ggtcaagaaa acatgtgtgt cctggcatta acatcaaggc gggtggggtg
       61 gggttgtttc tccaagaaag agaaacaccg tataggatac tatgagagat gagctatgat
      121 gtgaacttca gtttctccat ccgatgttgg atgtcaatag atcctgggac gcgtgcggcc
      181 gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
      241 ggccgggcga ccaaaggtcg cccgacgccc gggcggcctc agtgagcgag cgagcgcgca
      301 gctgcctgca ggacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
      361 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
      421 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
      481 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
      541 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
      601 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
      661 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
      721 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
      781 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
      841 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
      901 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
      961 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     1021 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     1081 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     1141 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     1201 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     1261 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     1321 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     1381 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     1441 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     1501 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     1561 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     1621 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     1681 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     1741 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     1801 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     1861 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     1921 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     1981 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     2041 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     2101 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     2161 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     2221 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     2281 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     2341 atgcggcatc agagcagatt gtactgagag tgcaccataa aattgtaaac gttaatattt
     2401 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa tagaccgaaa
     2461 tcggcaaaat cccttataaa tcaaaagaat agcccgagat agagttgagt gttgttccag
     2521 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     2581 tctatcaggg cgatggccca ctacgtgaac catcacccaa atcaagtttt ttggggtcga
     2641 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     2701 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaagg
     2761 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     2821 cgctacaggg cgcgtactat ggttgctttg acgtatgcgg tgtgaaatac cgcacagatg
     2881 cgtaaggaga aaataccgca tcaggcgccc ctgcaggcag ctgcgcgctc gctcgctcac
     2941 tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag
     3001 cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct gcggccgctc
     3061 ggtccgcacg tggttaccta caaaatcaga aggacaggga agggagcagt ggttcacgcc
     3121 tgtaatccca gcaatttggg aggccaaggt gggtagatca cctgagatta ggagttggag
     3181 accagcctgg ccaatatggt gaaaccccgt ctctaccaaa aaaacaaaaa ttagctgagc
     3241 ctggtcatgc atgcctggaa tcccaacaac tcgggaggct gaggcaggag aatcgcttga
     3301 acccaggagg cggagattgc agtgagccaa gattgtgcca ctgcactcca gcttggttcc
     3361 caatagaccc cgcaggccct acaggttgtc ttcccaactt gccccttgct ccataccacc
     3421 cccctccacc ccataatatt atagaaggac acctagtcag acaaaatgat gcaacttaat
     3481 tttattagga caaggctggt gggcactgga gtggcaactt ccagggccag gagaggcact
     3541 ggggaggggt cacagggatg ccacccgtag atctctcgag cagcgctcgg tatcgatgcg
     3601 gggaggcggc ccaaagggag atccgactcg tctgagggcg aaggcgaaga cgcggaagag
     3661 gccgcagagc cggcagcagg ccgcgggaag gaaggtccgc tggattgagg gccgaaggga
     3721 cgtagcagaa ggacgtcccg cgcagaatcc aggtggcaac ataggcgagc agccaaggaa
     3781 aggacgatga tttccccgac aacaccacgg aattgtcagt gcccaacagc cgagcccctg
     3841 tccagcagcg ggcaaggcag gcggcgatga gttccgccgt ggcaataggg agggggaaag
     3901 cgaaagtccc ggaaaggagc tgacaggtgg tggcaatgcc ccaaccagtg ggggttgcgt
     3961 cagcaaacac agtgcacacc acgccacgtt gcctgacaac gggccacaac tcctcataaa
     4021 gagacagcaa ccaggattta tacaaggagg agaaaatgaa agccatacgg gaagcaatag
     4081 catgatacaa aggcattaaa gcagcgtatc cacatagcgt aaaaggagca acatagttaa
     4141 gaataccagt caatctttca caaattttgt aatccagagg ttgattatcg ataagcttga
     4201 tatcgaattc tcagatccgc ctgttgatgt agctgctcag gtagtccagc acctcctggc
     4261 tgatgatgcc gttccaggcg gggtatctga tgctgccctc ggcgctgccc ttcagctgct
     4321 cgatgtgctg ccactcctcg atggggttgg tctcgtcctt cagggcgatc atctccttgc
     4381 tgatggggtc gtaggcgtag tacctggaca ccagggcgaa gtagtggtcg gggatggcgg
     4441 tgatctggtg ggtgtaggtg gtcctggcca cggcggaggc cctcttgtcg ctccagttgc
     4501 ccaccacgtt tgtcagctcg gtcaggccct tcatgctcag aaagctggtc atcaggtgcc
     4561 tgccgatgtg gctcttaggg ccgttcttga tagcgaagat ggggtagggg gcgttcttct
     4621 tcagggcctt gttgtagctg cgcaccaggt tgtccttcag cagctggtac tcctgcttgt
     4681 tgctgctgct gttgccggtc ctgttcactc tcttcagcac gggctcgctg ttcctcagga
     4741 actcgtccag gtacaccagg gggtcgatcc tgcctctggc gctgaaaaag tagatgtgcc
     4801 tggacacgct tgtcttggtc tcggtcacca ggcactgaat gatcacgccc aggtacttgt
     4861 tctgcaccag cttgaagctc ttggggtcca cgttcttgat gtcgctgaac ctgccgcagt
     4921 tgatgaatgt ggccaggaac aggaactggt acagggtctt ggtcttggtg aacctgctgg
     4981 tgtactcgaa gctgttcagg atcttctcgg tgatctccca gatgctctcg ccctcggaca
     5041 gcagggcctt cagcatcttc ttgctgtggc tgttgccctt gtcggcctcc tcgctgctct
     5101 cgaactgcag ctgcaggctg gacacgatgt cggtgatgtc gctctggtgc ttctggccgt
     5161 tgtaagggat gatggtgaac tcccaggcgg ggatcagctt cttcaggctg gcctccagga
     5221 tggtggcctt ctgggtcttg tacttgaact gcaggctctt gttcacgatg tcgaagctca
     5281 ggctgttgct gatgatggtg ttgtagctca tgaaggtggc cctcttgatg gcggtgccgt
     5341 tgtgggtgat catccagcac aggtaggtca gctcggcggc acagctggcg atcttctcgc
     5401 cgctgggcct ctcgaatctc tccacgaact gccgcaccag caccttgggg ggggtcttgc
     5461 acaggatgtc gaactggctc atcaccttcc tcttcttctt aggagccatg gtggccacgc
     5521 cagaagcaaa tgtaagcaat agatggctct gccctgactt ttatgcccag cccgagctcc
     5581 ttaagcaaac tggattcacg gtgaggcgta caggaaggca ccacggagtg attttggtca
     5641 tatttaatct gaaatgcctt gaacataatc acatgaactt ttattctcaa attggcaacg
     5701 ggcatttagg aggcattcac agattcactc acatggtgag tggtatttct caaaaaagac
     5761 agcatggcat aaatcggtag gaactgaact tggagactgc aactatctga gaactggtac
     5821 caacaacagg ctgaggaagc agagcgatga gtgatc
//
