LOCUS       sequence-461877-        5130 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0982h 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 5130)
  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 5130)
  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: AiP13465 -
            pAAV-AiE0982h-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5130
                     /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      596..605
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(602..604,605..607,608..1318)
                     /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(908..927)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1255..1276
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1328..1384
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1397..1462
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1493..1870
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(1957..1977)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2164..2275
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2393..2533
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2393..2522
                     /label=AAV2 ITR
     rep_origin      2608..3063
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2695..2714)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2905..2926
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3080..3099)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3199..3221
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3259..3277)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3345..3449
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3450..3518,3519..4310)
                     /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(3668..3687)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4481..5069
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4970..4989
                     /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 gcgtatctcg ttgtgatggg cttagccttc ccttctatca
      181 gaaggatgtt tctggctgca agtaattaaa ggccagactc aaagtggctt ccgcaatgaa
      241 ggaaatttat tagctcacaa agctaaaaag tgcaagaggc agtcaggttt caggtataat
      301 cagatcaggg ctccagcttg acttctattt tgagtctctc aactctgctc ctgtctgtgt
      361 ctgcttcatt tatatcctgg cttttttcac agtaaataac agctgaggta acttgacata
      421 caacgatttc cagaggaaga gagaacatct gtcctgacaa ctgtttggca agagcaagaa
      481 aatttcccag aaatcctccc cagagctcgg gctgggcata aaagtcaggg cagagccatc
      541 tattgcttac atttgcttct gggatccaga tctttcgaag ctagcgctac cggtcgccac
      601 catggtctcc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
      661 cggcgatgtc aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
      721 cggcaagctg accctgaagc tgatctgcac caccggcaag ctgcccgtgc cctggcccac
      781 cctcgtgacc accctgggct acggcgtgca gtgcttcgcc cgctaccccg accacatgaa
      841 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
      901 cttcaaagac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
      961 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     1021 caagctggag tacaactaca acagccacaa cgtctatatc accgccgaca agcagaagaa
     1081 cggcatcaag gccaacttca agatccgcca caacatcgag gacggcggcg tgcagctcgc
     1141 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     1201 ctacctgagc taccagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt
     1261 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaaggg
     1321 ctctggtgct accaacttct cactgttgaa acaggcaggg gatgtagagg agaatccagg
     1381 gcctggtgct agtggagact acaaagacca tgacggagat tataaagatc atgacatcga
     1441 ttacaaggat gacgatgaca agtccggact cagatctgga ggctccggag gcccagagcc
     1501 agcgaagtct gctcccgccc cgaaaaaggg ctccaagaag gcggtgacta aggcgcagaa
     1561 gaaaggcggc aagaagcgca agcgcagccg caaggagagc tattccatct atgtgtacaa
     1621 ggttctgaag caggtccacc ctgacaccgg catttcgtcc aaggccatgg gcatcatgaa
     1681 ttcgtttgtg aacgacattt tcgagcgcat cgcaggagag gcttcccgcc tggcgcatta
     1741 caacaagcgc tcgaccatca cctcccggga gatccagacg gccgtgcgcc tgctgctgcc
     1801 tggggagttg gccaagcacg ccgtgtccga gggtactaag gccatcacca agtacaccag
     1861 cgctaagtaa tgaggcgcgc cgcggccgcg aattcgatat cataatcaac ctctggatta
     1921 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     1981 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     2041 ctccttgtat aaatcctggt tagttcttgc cacggcggaa ctcatcgccg cctgccttgc
     2101 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggctc gagagatctt
     2161 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     2221 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     2281 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     2341 attgggaaga caatagcagg catgagatct cacgtgcgga ccgagcggcc gcaggaaccc
     2401 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
     2461 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc
     2521 agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     2581 caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg
     2641 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt
     2701 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     2761 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     2821 atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     2881 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact
     2941 ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa
     3001 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
     3061 ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     3121 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     3181 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     3241 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     3301 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     3361 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     3421 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     3481 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     3541 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     3601 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     3661 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     3721 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     3781 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     3841 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     3901 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     3961 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     4021 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     4081 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     4141 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     4201 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     4261 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     4321 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     4381 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     4441 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     4501 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     4561 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     4621 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     4681 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     4741 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     4801 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     4861 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     4921 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     4981 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     5041 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     5101 tggccttttg ctggcctttt gctcacatgt
//
