LOCUS       sequence-470293-        5830 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2054m 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 5830)
  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 5830)
  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: AiP1214 - pAAV-AiE2054m-minBG-FlpO-WPRE-HGHpA
FEATURES             Location/Qualifiers
     source          1..5830
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      628..637
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(634..639,640..660,661..1932)
                     /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    1957..2545
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2010..2030)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2428..2439)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2577..3053
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2776..2795)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3093..3233
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3093..3222
                     /label=AAV2 ITR
     rep_origin      3308..3763
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3395..3414)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3605..3626
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3780..3799)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3899..3921
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3959..3977)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4045..4149
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4150..4218,4219..5010)
                     /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(4368..4387)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5181..5769
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5670..5689
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtcacaaa tgtaatgcag tcgccatctg ccatttcagc
      181 acctgccctt cagggtgttt atggctgata ttttaaacat acagtaaatg cagaaaggaa
      241 attttataca gcataaccaa caagtatgca taaaaattca cccctgtaac taaaattctt
      301 ttacatatct acttaatctt atttttaaga aaacactttt tttttcttct atagcaataa
      361 aatgctacat ttaaaatcaa tctgtgcata gctcactatc atggattaaa aacagtagct
      421 ggtgtggcat ttttagcaga ctgtacaaaa tggaaatgtc accttttctt ttgggtgaaa
      481 gtactttgca tttcccaaga ataaaataat ctgagtttaa cattagcttt aagaagaaaa
      541 attctgtgtt aaactaaaag ttcgagctcg ggctgggcat aaaagtcagg gcagagccat
      601 ctattgctta catttgcttc tggcgtggcc accatggctc ctaagaagaa gaggaaggtg
      661 atgagccagt tcgacatcct gtgcaagacc ccccccaagg tgctggtgcg gcagttcgtg
      721 gagagattcg agaggcccag cggcgagaag atcgccagct gtgccgccga gctgacctac
      781 ctgtgctgga tgatcaccca caacggcacc gccatcaaga gggccacctt catgagctac
      841 aacaccatca tcagcaacag cctgagcttc gacatcgtga acaagagcct gcagttcaag
      901 tacaagaccc agaaggccac catcctggag gccagcctga agaagctgat ccccgcctgg
      961 gagttcacca tcatccctta caacggccag aagcaccaga gcgacatcac cgacatcgtg
     1021 tccagcctgc agctgcagtt cgagagcagc gaggaggccg acaagggcaa cagccacagc
     1081 aagaagatgc tgaaggccct gctgtccgag ggcgagagca tctgggagat caccgagaag
     1141 atcctgaaca gcttcgagta caccagcagg ttcaccaaga ccaagaccct gtaccagttc
     1201 ctgttcctgg ccacattcat caactgcggc aggttcagcg acatcaagaa cgtggacccc
     1261 aagagcttca agctggtgca gaacaagtac ctgggcgtga tcattcagtg cctggtgacc
     1321 gagaccaaga caagcgtgtc caggcacatc tactttttca gcgccagagg caggatcgac
     1381 cccctggtgt acctggacga gttcctgagg aacagcgagc ccgtgctgaa gagagtgaac
     1441 aggaccggca acagcagcag caacaagcag gagtaccagc tgctgaagga caacctggtg
     1501 cgcagctaca acaaggccct gaagaagaac gccccctacc ccatcttcgc tatcaagaac
     1561 ggccctaaga gccacatcgg caggcacctg atgaccagct ttctgagcat gaagggcctg
     1621 accgagctga caaacgtggt gggcaactgg agcgacaaga gggcctccgc cgtggccagg
     1681 accacctaca cccaccagat caccgccatc cccgaccact acttcgccct ggtgtccagg
     1741 tactacgcct acgaccccat cagcaaggag atgatcgccc tgaaggacga gaccaacccc
     1801 atcgaggagt ggcagcacat cgagcagctg aagggcagcg ccgagggcag catcagatac
     1861 cccgcctgga acggcatcat cagccaggag gtgctggact acctgagcag ctacatcaac
     1921 aggcggatct gagaattcga tatcaagctt atcgataatc aacctctgga ttacaaaatt
     1981 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     2041 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     2101 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     2161 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     2221 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     2281 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     2341 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct atgttgccac ctggattctg
     2401 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     2461 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     2521 atctcccttt gggccgcctc cccgcatcga taccgagcgc tgctcgagag atctacgggt
     2581 ggcatccctg tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc
     2641 ccaccagcct tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct
     2701 ataatattat ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct
     2761 gtagggcctg cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca
     2821 ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg
     2881 gattccaggc atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt
     2941 tcaccatatt ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc
     3001 ctcccaaatt gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt
     3061 tgtaggtaac cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac
     3121 tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc
     3181 gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct
     3241 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa
     3301 ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     3361 gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt
     3421 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     3481 cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt
     3541 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     3601 aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt
     3661 gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa
     3721 aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt
     3781 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     3841 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     3901 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     3961 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     4021 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4081 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4141 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     4201 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     4261 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     4321 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     4381 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     4441 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     4501 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     4561 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     4621 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     4681 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     4741 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     4801 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     4861 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     4921 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     4981 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     5041 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5101 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5161 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5221 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5281 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     5341 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5401 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5461 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5521 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     5581 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     5641 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     5701 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     5761 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     5821 gctcacatgt
//
