LOCUS       sequence-474057-        5409 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0098h 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 5409)
  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 5409)
  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: AiP11544 -
            pAAV-hsA2-AiE0098h-minRho-SYFP2-WPR...
FEATURES             Location/Qualifiers
     source          1..5409
                     /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      1468..1477
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1474..1476,1477..1479,1480..2190)
                     /codon_start=1
                     /product="monomeric optimized variant of EYFP, identical to
                     mVenus but with L68V (Kremers et al., 2006)"
                     /label=SYFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1519..1540)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1780..1799)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2127..2148
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2258..2278)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2465..2576
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2887..3342
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2974..2993)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3184..3205
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3359..3378)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3478..3500
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3538..3556)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3624..3728
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3729..3797,3798..4589)
                     /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(3947..3966)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4760..5348
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5249..5268
                     /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 gcggccgcaa cgcgtttaga acaatggctg gcccatagta aatgccgtgt
      181 tagtgtgtta gttgctgttc ttccacgtca gaagaggcac agacaaatta ccaccaggtg
      241 gcgctcagag tctgcggagg catcacaaca gccctgaatt tgaatcctgc tctgccactg
      301 cctagttgag accttttact acctgactag ctgtttgtgt attttaggtg tttgttttga
      361 ctgacaggcc gggcccctgc tcatccctga aaactagcat cggggaatga catacagttt
      421 gataacataa gatgcttagg ttttttggct caagtgagat gttcagagct aatagccctg
      481 cgagctgagt tttagctgtg atctgcacct tgagatatcc gttatgtctg gatcaggaaa
      541 aagggaaaag aatgagagct tatttctaga aaggtttccg atgagcagca tttgtcattg
      601 aaggggaaga tgggtataat tagcttgtca gtctagtttg ggctgagcag gctgtctctg
      661 ctaagaaaga tggactaact gccagggcca ctgacaggca gtcgatagcg cagcaattaa
      721 tgtcataaat aattccagaa ccaattacat caaaatggac tcagagcttt aaaaaggtac
      781 agaattctct tattcttcct gctaacagca agaaacagct ttcatttttc tctgtaatta
      841 cttttcattg tatatagcaa tacagctgag cttttctgtc catatttaag ccttgtcttt
      901 ctccaagcct cttcccccac caccagcctc cctccttcac cccacaggat aaggctgtta
      961 aagttgttct tccctgccag agcagtgggg tgaatgtata atcttttaac ttggctgttg
     1021 caggagaact ccagctacac ctaaatgatt aagagccaag tttttgaaaa tgtcatggac
     1081 aatgacagca cagtgcccca aacaatggca tttggaaatg aagacatttg tgaaataaat
     1141 ggatttgtct tattatttta aaggttgctg tgactttgct tacactgaat ttattgattt
     1201 ttcccccttc tccttctcct tcctcttctt gttctcctcc tccttctgta cagttgcagg
     1261 catagaagct gggccgaatt cgatatcata atcaaccata ggtaccgagc tcgggattca
     1321 gccgggagct tagggagggg aggtcacttc ataagggcct ggggggggag ttggagccac
     1381 gagtcgtcca gccggagccc cgtgtggctg agctccggcc tcagaagcat ccccgggttg
     1441 gatccttcga agctagcgct accggtcgcc accatggtga gcaagggcga ggagctgttc
     1501 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     1561 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gctgatctgc
     1621 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctggg ctacggcgtg
     1681 cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1741 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1801 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1861 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     1921 aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc
     1981 cacaacatcg aggacggcgg cgtgcagctc gccgaccact accagcagaa cacccccatc
     2041 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc caagctgagc
     2101 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     2161 atcactctcg gcatggacga gctgtacaag taagtcgaca tcataatcaa cctctggatt
     2221 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     2281 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     2341 cctccttgta taaatcctgg ttagttcttg ccacggcgga actcatcgcc gcctgccttg
     2401 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggct cgagagatct
     2461 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     2521 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     2581 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     2641 gattgggaag acaatagcag gcatgagatc tcacgtgcgg accgagcggc cgcaggaacc
     2701 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
     2761 accaaaggtc gcccggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg
     2821 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac
     2881 catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     2941 tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc
     3001 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     3061 gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta
     3121 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     3181 atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg
     3241 atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa
     3301 aatttaacgg gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta
     3361 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     3421 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     3481 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
     3541 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     3601 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     3661 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     3721 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     3781 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     3841 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     3901 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     3961 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     4021 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     4081 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     4141 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     4201 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     4261 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     4321 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     4381 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     4441 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     4501 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     4561 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     4621 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     4681 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     4741 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     4801 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     4861 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     4921 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     4981 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     5041 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     5101 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     5161 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     5221 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     5281 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     5341 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     5401 ctcacatgt
//
