LOCUS       sequence-461876-        5120 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0973h 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 5120)
  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 5120)
  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: AiP13457 -
            pAAV-AiE0973h-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5120
                     /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      586..595
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(592..594,595..597,598..1308)
                     /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(898..917)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1245..1266
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1318..1374
                     /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             1387..1452
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1483..1860
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(1947..1967)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2154..2265
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2383..2523
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2383..2512
                     /label=AAV2 ITR
     rep_origin      2598..3053
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2685..2704)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2895..2916
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3070..3089)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3189..3211
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3249..3267)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3335..3439
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3440..3508,3509..4300)
                     /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(3658..3677)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4471..5059
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4960..4979
                     /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 gcgttgatag aaaacccagc gagagggagg gttgtctgga
      181 gccatcaggg aaggctttgt gaaaggcttg tggcttaacc tggccttctg cgatgggaag
      241 ctttgggaag agcagagcag agaagattgg gtagtaaact gcagggataa attgcaggtt
      301 gcatgttgaa ggtaaagcac agactgtctt tgtatgggtt ggggaaatga ggagcgaaag
      361 gccagatggt gatgagcatg gagatgggga aattttgaag agagaactga aggatatata
      421 catatgtgaa cacagaatca ctgaaagaga aggatgttat tttaagagaa tttcgttcag
      481 ctcagccttt cagagctcgg gctgggcata aaagtcaggg cagagccatc tattgcttac
      541 atttgcttct gggatccaga tctttcgaag ctagcgctac cggtcgccac catggtctcc
      601 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgatgtc
      661 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
      721 accctgaagc tgatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
      781 accctgggct acggcgtgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac
      841 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaagac
      901 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
      961 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     1021 tacaactaca acagccacaa cgtctatatc accgccgaca agcagaagaa cggcatcaag
     1081 gccaacttca agatccgcca caacatcgag gacggcggcg tgcagctcgc cgaccactac
     1141 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     1201 taccagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     1261 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaaggg ctctggtgct
     1321 accaacttct cactgttgaa acaggcaggg gatgtagagg agaatccagg gcctggtgct
     1381 agtggagact acaaagacca tgacggagat tataaagatc atgacatcga ttacaaggat
     1441 gacgatgaca agtccggact cagatctgga ggctccggag gcccagagcc agcgaagtct
     1501 gctcccgccc cgaaaaaggg ctccaagaag gcggtgacta aggcgcagaa gaaaggcggc
     1561 aagaagcgca agcgcagccg caaggagagc tattccatct atgtgtacaa ggttctgaag
     1621 caggtccacc ctgacaccgg catttcgtcc aaggccatgg gcatcatgaa ttcgtttgtg
     1681 aacgacattt tcgagcgcat cgcaggagag gcttcccgcc tggcgcatta caacaagcgc
     1741 tcgaccatca cctcccggga gatccagacg gccgtgcgcc tgctgctgcc tggggagttg
     1801 gccaagcacg ccgtgtccga gggtactaag gccatcacca agtacaccag cgctaagtaa
     1861 tgaggcgcgc cgcggccgcg aattcgatat cataatcaac ctctggatta caaaatttgt
     1921 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     1981 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     2041 aaatcctggt tagttcttgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     2101 acaggggctc ggctgttggg cactgacaat tccgtggctc gagagatctt cgactgtgcc
     2161 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     2221 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     2281 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     2341 caatagcagg catgagatct cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg
     2401 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
     2461 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc
     2521 aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
     2581 gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     2641 tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt
     2701 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     2761 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga
     2821 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     2881 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg
     2941 ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct
     3001 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg
     3061 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     3121 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     3181 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     3241 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     3301 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     3361 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     3421 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     3481 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     3541 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     3601 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     3661 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     3721 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     3781 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     3841 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     3901 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     3961 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     4021 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     4081 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     4141 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     4201 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     4261 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     4321 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     4381 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     4441 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     4501 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     4561 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     4621 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     4681 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     4741 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     4801 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     4861 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     4921 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     4981 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     5041 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     5101 ctggcctttt gctcacatgt
//
