LOCUS       sequence-470276-        5839 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0337m 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 5839)
  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 5839)
  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: AiP1098 - pAAV-AiE0337m-minBG-FlpO-WPRE-HGHpA
FEATURES             Location/Qualifiers
     source          1..5839
                     /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      637..646
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(643..648,649..669,670..1941)
                     /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    1966..2554
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2019..2039)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2437..2448)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2586..3062
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2785..2804)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3102..3242
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3102..3231
                     /label=AAV2 ITR
     rep_origin      3317..3772
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3404..3423)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3614..3635
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3789..3808)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3908..3930
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3968..3986)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4054..4158
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4159..4227,4228..5019)
                     /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(4377..4396)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5190..5778
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5679..5698
                     /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 gcgtttctgt ccctcagaac tgtgaaataa aatacatttc
      181 tgttgcttaa accacccagt ctaggctatc ttgttatagc aacaagaaaa cactgatata
      241 cctggtcaat tcaaatgagc tgagtaacct tcattcggat ctccaaattt tcatttgcat
      301 ttttaaagct tcctctgaat actccaggtg caatctcctc atgatctcgt ggcttgctgc
      361 tcacttgtga aactcaacct tttctatcta caaaaggacg tctctatctg tactctgagt
      421 cagaacccta ggagttgttc ttgatgtcac attctcctca cgcccacttt aaaccctcct
      481 catcccattg tttcattgtc tttcatattt ttcagtgctg cagaggaaca catttctatc
      541 attaatgcac catgctcaga catcacttca gtgagctcgg gctgggcata aaagtcaggg
      601 cagagccatc tattgcttac atttgcttct ggcgtggcca ccatggctcc taagaagaag
      661 aggaaggtga tgagccagtt cgacatcctg tgcaagaccc cccccaaggt gctggtgcgg
      721 cagttcgtgg agagattcga gaggcccagc ggcgagaaga tcgccagctg tgccgccgag
      781 ctgacctacc tgtgctggat gatcacccac aacggcaccg ccatcaagag ggccaccttc
      841 atgagctaca acaccatcat cagcaacagc ctgagcttcg acatcgtgaa caagagcctg
      901 cagttcaagt acaagaccca gaaggccacc atcctggagg ccagcctgaa gaagctgatc
      961 cccgcctggg agttcaccat catcccttac aacggccaga agcaccagag cgacatcacc
     1021 gacatcgtgt ccagcctgca gctgcagttc gagagcagcg aggaggccga caagggcaac
     1081 agccacagca agaagatgct gaaggccctg ctgtccgagg gcgagagcat ctgggagatc
     1141 accgagaaga tcctgaacag cttcgagtac accagcaggt tcaccaagac caagaccctg
     1201 taccagttcc tgttcctggc cacattcatc aactgcggca ggttcagcga catcaagaac
     1261 gtggacccca agagcttcaa gctggtgcag aacaagtacc tgggcgtgat cattcagtgc
     1321 ctggtgaccg agaccaagac aagcgtgtcc aggcacatct actttttcag cgccagaggc
     1381 aggatcgacc ccctggtgta cctggacgag ttcctgagga acagcgagcc cgtgctgaag
     1441 agagtgaaca ggaccggcaa cagcagcagc aacaagcagg agtaccagct gctgaaggac
     1501 aacctggtgc gcagctacaa caaggccctg aagaagaacg ccccctaccc catcttcgct
     1561 atcaagaacg gccctaagag ccacatcggc aggcacctga tgaccagctt tctgagcatg
     1621 aagggcctga ccgagctgac aaacgtggtg ggcaactgga gcgacaagag ggcctccgcc
     1681 gtggccagga ccacctacac ccaccagatc accgccatcc ccgaccacta cttcgccctg
     1741 gtgtccaggt actacgccta cgaccccatc agcaaggaga tgatcgccct gaaggacgag
     1801 accaacccca tcgaggagtg gcagcacatc gagcagctga agggcagcgc cgagggcagc
     1861 atcagatacc ccgcctggaa cggcatcatc agccaggagg tgctggacta cctgagcagc
     1921 tacatcaaca ggcggatctg agaattcgat atcaagctta tcgataatca acctctggat
     1981 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     2041 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     2101 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     2161 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     2221 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     2281 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     2341 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgccta tgttgccacc
     2401 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     2461 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     2521 acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgagcgct gctcgagaga
     2581 tctacgggtg gcatccctgt gacccctccc cagtgcctct cctggccctg gaagttgcca
     2641 ctccagtgcc caccagcctt gtcctaataa aattaagttg catcattttg tctgactagg
     2701 tgtccttcta taatattatg gggtggaggg gggtggtatg gagcaagggg caagttggga
     2761 agacaacctg tagggcctgc ggggtctatt gggaaccaag ctggagtgca gtggcacaat
     2821 cttggctcac tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct cagcctcccg
     2881 agttgttggg attccaggca tgcatgacca ggctcagcta atttttgttt ttttggtaga
     2941 gacggggttt caccatattg gccaggctgg tctccaactc ctaatctcag gtgatctacc
     3001 caccttggcc tcccaaattg ctgggattac aggcgtgaac cactgctccc ttccctgtcc
     3061 ttctgatttt gtaggtaacc acgtgcggac cgagcggccg caggaacccc tagtgatgga
     3121 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
     3181 ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca
     3241 ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg
     3301 tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3361 acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc
     3421 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     3481 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat
     3541 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     3601 acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc
     3661 tattcttttg atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg
     3721 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc
     3781 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     3841 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     3901 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     3961 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     4021 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     4081 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     4141 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     4201 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     4261 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     4321 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     4381 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     4441 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     4501 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     4561 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     4621 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     4681 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     4741 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     4801 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     4861 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     4921 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     4981 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     5041 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     5101 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5161 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5221 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5281 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     5341 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     5401 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     5461 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     5521 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     5581 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     5641 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     5701 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     5761 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     5821 tggccttttg ctcacatgt
//
