LOCUS       sequence-470289-        6019 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0455m 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 6019)
  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 6019)
  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: AiP1189 - pAAV-AiE0455m-minBG-FlpO-WPRE-HGHpA
FEATURES             Location/Qualifiers
     source          1..6019
                     /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      817..826
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(823..828,829..849,850..2121)
                     /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    2146..2734
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2199..2219)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2617..2628)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2766..3242
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2965..2984)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3282..3422
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3282..3411
                     /label=AAV2 ITR
     rep_origin      3497..3952
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3584..3603)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3794..3815
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3969..3988)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4088..4110
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4148..4166)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4234..4338
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4339..4407,4408..5199)
                     /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(4557..4576)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5370..5958
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5859..5878
                     /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 gcgtcggagc ccagagcact ttagcaataa agctgtaaat
      181 gaaaagaaat gcattcaagc tgttaacctt atagagatgg aaaattaatt atcataataa
      241 tggcttccac acttctatcc aatgctggct tctggaggct ctactggaga cagaattcac
      301 tcttgtgttt atttattcac tcatcccttt ggtggctccc tgcagcacct ggtctgtgta
      361 tgtcaggagt caggctcact gggaaggatg agaaggagaa gcagacaaga tgcagagtga
      421 aaaggtgttg ctaccacaca atgtcctttc ccagcctgtc attcattcta aactaccagt
      481 ccctgaaaca aactcaatga ggctctagac gggcttgtat accctccgtc tagacgggtg
      541 tccaattgag cgccattatc cctacagaca cgcccttccc caccccatga aaggactgtg
      601 ccatttgcca atgacactca gcacacagca gccacagctt tgctgcttct gtccctacat
      661 tgggactccc agctgggaca catggcagac agtgcaatct gagtggtgtg agggacctaa
      721 agttattggt gtatgacagc tttcccctta aggagctcgg gctgggcata aaagtcaggg
      781 cagagccatc tattgcttac atttgcttct ggcgtggcca ccatggctcc taagaagaag
      841 aggaaggtga tgagccagtt cgacatcctg tgcaagaccc cccccaaggt gctggtgcgg
      901 cagttcgtgg agagattcga gaggcccagc ggcgagaaga tcgccagctg tgccgccgag
      961 ctgacctacc tgtgctggat gatcacccac aacggcaccg ccatcaagag ggccaccttc
     1021 atgagctaca acaccatcat cagcaacagc ctgagcttcg acatcgtgaa caagagcctg
     1081 cagttcaagt acaagaccca gaaggccacc atcctggagg ccagcctgaa gaagctgatc
     1141 cccgcctggg agttcaccat catcccttac aacggccaga agcaccagag cgacatcacc
     1201 gacatcgtgt ccagcctgca gctgcagttc gagagcagcg aggaggccga caagggcaac
     1261 agccacagca agaagatgct gaaggccctg ctgtccgagg gcgagagcat ctgggagatc
     1321 accgagaaga tcctgaacag cttcgagtac accagcaggt tcaccaagac caagaccctg
     1381 taccagttcc tgttcctggc cacattcatc aactgcggca ggttcagcga catcaagaac
     1441 gtggacccca agagcttcaa gctggtgcag aacaagtacc tgggcgtgat cattcagtgc
     1501 ctggtgaccg agaccaagac aagcgtgtcc aggcacatct actttttcag cgccagaggc
     1561 aggatcgacc ccctggtgta cctggacgag ttcctgagga acagcgagcc cgtgctgaag
     1621 agagtgaaca ggaccggcaa cagcagcagc aacaagcagg agtaccagct gctgaaggac
     1681 aacctggtgc gcagctacaa caaggccctg aagaagaacg ccccctaccc catcttcgct
     1741 atcaagaacg gccctaagag ccacatcggc aggcacctga tgaccagctt tctgagcatg
     1801 aagggcctga ccgagctgac aaacgtggtg ggcaactgga gcgacaagag ggcctccgcc
     1861 gtggccagga ccacctacac ccaccagatc accgccatcc ccgaccacta cttcgccctg
     1921 gtgtccaggt actacgccta cgaccccatc agcaaggaga tgatcgccct gaaggacgag
     1981 accaacccca tcgaggagtg gcagcacatc gagcagctga agggcagcgc cgagggcagc
     2041 atcagatacc ccgcctggaa cggcatcatc agccaggagg tgctggacta cctgagcagc
     2101 tacatcaaca ggcggatctg agaattcgat atcaagctta tcgataatca acctctggat
     2161 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     2221 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     2281 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     2341 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     2401 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     2461 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     2521 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgccta tgttgccacc
     2581 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     2641 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     2701 acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgagcgct gctcgagaga
     2761 tctacgggtg gcatccctgt gacccctccc cagtgcctct cctggccctg gaagttgcca
     2821 ctccagtgcc caccagcctt gtcctaataa aattaagttg catcattttg tctgactagg
     2881 tgtccttcta taatattatg gggtggaggg gggtggtatg gagcaagggg caagttggga
     2941 agacaacctg tagggcctgc ggggtctatt gggaaccaag ctggagtgca gtggcacaat
     3001 cttggctcac tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct cagcctcccg
     3061 agttgttggg attccaggca tgcatgacca ggctcagcta atttttgttt ttttggtaga
     3121 gacggggttt caccatattg gccaggctgg tctccaactc ctaatctcag gtgatctacc
     3181 caccttggcc tcccaaattg ctgggattac aggcgtgaac cactgctccc ttccctgtcc
     3241 ttctgatttt gtaggtaacc acgtgcggac cgagcggccg caggaacccc tagtgatgga
     3301 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
     3361 ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca
     3421 ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg
     3481 tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3541 acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc
     3601 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     3661 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat
     3721 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     3781 acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc
     3841 tattcttttg atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg
     3901 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc
     3961 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     4021 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     4081 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     4141 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     4201 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     4261 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     4321 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     4381 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     4441 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     4501 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     4561 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     4621 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     4681 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     4741 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     4801 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     4861 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     4921 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     4981 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     5041 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     5101 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     5161 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     5221 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     5281 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5341 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5401 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5461 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     5521 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     5581 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     5641 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     5701 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     5761 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     5821 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     5881 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     5941 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6001 tggccttttg ctcacatgt
//
