LOCUS       sequence-466279-        5163 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2114m 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 5163)
  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 5163)
  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: AiP1633 - pAAV-AiE2114m-minBG-FlpO-HGHpA
FEATURES             Location/Qualifiers
     source          1..5163
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   53..182
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      650..659
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(656..661,662..682,683..1954)
                     /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"
     polyA_signal    1962..2438
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2161..2180)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2478..2618
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2478..2607
                     /label=AAV2 ITR
     rep_origin      2693..3148
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2780..2799)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2990..3011
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3165..3184)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3284..3306
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3344..3362)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3430..3534
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3535..3603,3604..4395)
                     /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(3753..3772)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4566..5154
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5055..5074
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gtcctgcagg
       61 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcgt cgggcgacct ttggtcgccc
      121 ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc
      181 ctgcggccgc acgcgtttgg catttgacat ttacctgaag tctgtcttgt gccgggtcct
      241 gtccccaggt tgggtctaag tctcacgctg gatctcagtg ctcacagctt cctccagaac
      301 atacctgctc tggcattatc tttcctgagc agatgctaga acttggctca gagtgatgaa
      361 gaggaagtgt gtcctctggc taaagtccag gggttgcaga ggggagtgtg tgagatattt
      421 taatacatac ctatcttgag aaggtagcag gggtgttaaa gcatatgtaa tcatccaggc
      481 ttcttcatat ctcaagaaat ctagcatagc acaaagatgg gctagatgta cccaaaggtg
      541 catgcaaaac gtaagcagct tcttgtttcc tatttcactg agtttgagct cgggctgggc
      601 ataaaagtca gggcagagcc atctattgct tacatttgct tctggcgtgg ccaccatggc
      661 tcctaagaag aagaggaagg tgatgagcca gttcgacatc ctgtgcaaga ccccccccaa
      721 ggtgctggtg cggcagttcg tggagagatt cgagaggccc agcggcgaga agatcgccag
      781 ctgtgccgcc gagctgacct acctgtgctg gatgatcacc cacaacggca ccgccatcaa
      841 gagggccacc ttcatgagct acaacaccat catcagcaac agcctgagct tcgacatcgt
      901 gaacaagagc ctgcagttca agtacaagac ccagaaggcc accatcctgg aggccagcct
      961 gaagaagctg atccccgcct gggagttcac catcatccct tacaacggcc agaagcacca
     1021 gagcgacatc accgacatcg tgtccagcct gcagctgcag ttcgagagca gcgaggaggc
     1081 cgacaagggc aacagccaca gcaagaagat gctgaaggcc ctgctgtccg agggcgagag
     1141 catctgggag atcaccgaga agatcctgaa cagcttcgag tacaccagca ggttcaccaa
     1201 gaccaagacc ctgtaccagt tcctgttcct ggccacattc atcaactgcg gcaggttcag
     1261 cgacatcaag aacgtggacc ccaagagctt caagctggtg cagaacaagt acctgggcgt
     1321 gatcattcag tgcctggtga ccgagaccaa gacaagcgtg tccaggcaca tctacttttt
     1381 cagcgccaga ggcaggatcg accccctggt gtacctggac gagttcctga ggaacagcga
     1441 gcccgtgctg aagagagtga acaggaccgg caacagcagc agcaacaagc aggagtacca
     1501 gctgctgaag gacaacctgg tgcgcagcta caacaaggcc ctgaagaaga acgcccccta
     1561 ccccatcttc gctatcaaga acggccctaa gagccacatc ggcaggcacc tgatgaccag
     1621 ctttctgagc atgaagggcc tgaccgagct gacaaacgtg gtgggcaact ggagcgacaa
     1681 gagggcctcc gccgtggcca ggaccaccta cacccaccag atcaccgcca tccccgacca
     1741 ctacttcgcc ctggtgtcca ggtactacgc ctacgacccc atcagcaagg agatgatcgc
     1801 cctgaaggac gagaccaacc ccatcgagga gtggcagcac atcgagcagc tgaagggcag
     1861 cgccgagggc agcatcagat accccgcctg gaacggcatc atcagccagg aggtgctgga
     1921 ctacctgagc agctacatca acaggcggat ctgagaatta cgggtggcat ccctgtgacc
     1981 cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc agccttgtcc
     2041 taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat attatggggt
     2101 ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg gcctgcgggg
     2161 tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca atctccgcct
     2221 cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc caggcatgca
     2281 tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc atattggcca
     2341 ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc aaattgctgg
     2401 gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttgtag gtaaccacgt
     2461 gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc
     2521 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc
     2581 ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct
     2641 ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc
     2701 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     2761 gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     2821 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     2881 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     2941 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     3001 ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt
     3061 ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     3121 tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat
     3181 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     3241 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     3301 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     3361 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     3421 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     3481 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     3541 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     3601 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     3661 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     3721 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     3781 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     3841 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     3901 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     3961 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     4021 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     4081 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     4141 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     4201 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     4261 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     4321 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     4381 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     4441 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     4501 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     4561 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     4621 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     4681 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     4741 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     4801 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     4861 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     4921 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     4981 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     5041 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     5101 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     5161 agc
//
