LOCUS       sequence-470277-        6383 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0338m 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 6383)
  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 6383)
  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: AiP1099 - pAAV-AiE0338m-minBG-FlpO-WPRE-HGHpA
FEATURES             Location/Qualifiers
     source          1..6383
                     /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      1181..1190
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1187..1192,1193..1213,1214..2485)
                     /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    2510..3098
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2563..2583)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2981..2992)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3130..3606
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3329..3348)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3646..3786
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3646..3775
                     /label=AAV2 ITR
     rep_origin      3861..4316
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3948..3967)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4158..4179
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4333..4352)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4452..4474
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4512..4530)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4598..4702
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4703..4771,4772..5563)
                     /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(4921..4940)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5734..6322
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6223..6242
                     /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 gcgtttccct ttagctcctt ggttactttc tctagctcct
      181 cccttgggag ccctgcgatt tatccatgta atgttttgga atttttacca agtctgatac
      241 atataaagat aaaacaaaac aaaatatttt cttatgacta gtccaagcat gaagaaggac
      301 aaggctaagc agaagtttat caaagcagca ttcaaattag aaggtagtca aaacaaaaat
      361 acaatgaatc agaacagaaa aagaacctga atctcaaagg tttagggaat tctgctgaga
      421 tggtaaaagt gtaatgggtt gttaggaaca tatggttcaa agtctcttgc tctctctttt
      481 catttctgtt agttttactt aagagatagg tccaacaaaa tcaagttata agtataattg
      541 tctcagtaag aagatgacta aattgataag gatccgaaga gtgaataaag atgctgacga
      601 catggccaaa gtaaaatgga ttaggctgtc ttcatgctag gactttaaat tgatgtgtac
      661 aaaattaaga agtgctgttg caacagtgtt tcaaacatcc tagccaggtg atgtttattc
      721 tggggttaaa cacaagaagg gtaaatgtag gcagatggat tctgaagaaa tcaaaagctt
      781 gtttaaatgc aaatttttct acagagaaga agcattcaga gaagaaatga tatctaaata
      841 aattttcttt cagcagaatg ctttcttttt caggtttgaa agaggcaaac tcaggtgtct
      901 aatggagatt tactaaagaa ctaaaactgg gtacctttgc aaacggaatt tcagtgcctc
      961 taaaagccta ctctctgaag cactttactt agctgtggag ctgaacctgc aatcaaatta
     1021 caatcttctt tgaattttgc ataccaatct tgacttcaag atgagttgat acttcaagac
     1081 ataaatgtac aatttggtct ctctcttatg atccacgagc tcgggctggg cataaaagtc
     1141 agggcagagc catctattgc ttacatttgc ttctggcgtg gccaccatgg ctcctaagaa
     1201 gaagaggaag gtgatgagcc agttcgacat cctgtgcaag acccccccca aggtgctggt
     1261 gcggcagttc gtggagagat tcgagaggcc cagcggcgag aagatcgcca gctgtgccgc
     1321 cgagctgacc tacctgtgct ggatgatcac ccacaacggc accgccatca agagggccac
     1381 cttcatgagc tacaacacca tcatcagcaa cagcctgagc ttcgacatcg tgaacaagag
     1441 cctgcagttc aagtacaaga cccagaaggc caccatcctg gaggccagcc tgaagaagct
     1501 gatccccgcc tgggagttca ccatcatccc ttacaacggc cagaagcacc agagcgacat
     1561 caccgacatc gtgtccagcc tgcagctgca gttcgagagc agcgaggagg ccgacaaggg
     1621 caacagccac agcaagaaga tgctgaaggc cctgctgtcc gagggcgaga gcatctggga
     1681 gatcaccgag aagatcctga acagcttcga gtacaccagc aggttcacca agaccaagac
     1741 cctgtaccag ttcctgttcc tggccacatt catcaactgc ggcaggttca gcgacatcaa
     1801 gaacgtggac cccaagagct tcaagctggt gcagaacaag tacctgggcg tgatcattca
     1861 gtgcctggtg accgagacca agacaagcgt gtccaggcac atctactttt tcagcgccag
     1921 aggcaggatc gaccccctgg tgtacctgga cgagttcctg aggaacagcg agcccgtgct
     1981 gaagagagtg aacaggaccg gcaacagcag cagcaacaag caggagtacc agctgctgaa
     2041 ggacaacctg gtgcgcagct acaacaaggc cctgaagaag aacgccccct accccatctt
     2101 cgctatcaag aacggcccta agagccacat cggcaggcac ctgatgacca gctttctgag
     2161 catgaagggc ctgaccgagc tgacaaacgt ggtgggcaac tggagcgaca agagggcctc
     2221 cgccgtggcc aggaccacct acacccacca gatcaccgcc atccccgacc actacttcgc
     2281 cctggtgtcc aggtactacg cctacgaccc catcagcaag gagatgatcg ccctgaagga
     2341 cgagaccaac cccatcgagg agtggcagca catcgagcag ctgaagggca gcgccgaggg
     2401 cagcatcaga taccccgcct ggaacggcat catcagccag gaggtgctgg actacctgag
     2461 cagctacatc aacaggcgga tctgagaatt cgatatcaag cttatcgata atcaacctct
     2521 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     2581 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     2641 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     2701 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     2761 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     2821 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     2881 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctatgttgc
     2941 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     3001 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     3061 tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgag cgctgctcga
     3121 gagatctacg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc cctggaagtt
     3181 gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat tttgtctgac
     3241 taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa ggggcaagtt
     3301 gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag tgcagtggca
     3361 caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct gcctcagcct
     3421 cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt gtttttttgg
     3481 tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc tcaggtgatc
     3541 tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc tcccttccct
     3601 gtccttctga ttttgtaggt aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga
     3661 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     3721 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     3781 tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     3841 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     3901 ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt
     3961 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     4021 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     4081 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     4141 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc
     4201 gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga
     4261 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     4321 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     4381 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     4441 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     4501 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     4561 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     4621 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4681 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     4741 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     4801 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     4861 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     4921 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     4981 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     5041 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     5101 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     5161 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     5221 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     5281 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     5341 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     5401 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     5461 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     5521 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     5581 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     5641 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     5701 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5761 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5821 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5881 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5941 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     6001 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     6061 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     6121 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     6181 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6241 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6301 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     6361 ttgctggcct tttgctcaca tgt
//
