LOCUS       sequence-470711-        5300 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE1019h 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 5300)
  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 5300)
  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: AiP13611 -
            pAAV-AiE1019h-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5300
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      126..135
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(132..134,135..137,138..848)
                     /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(438..457)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     785..806
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             858..914
                     /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             927..992
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1023..1400
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(1487..1507)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1694..1805
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1923..2063
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1923..2052
                     /label=AAV2 ITR
     rep_origin      2138..2593
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2225..2244)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2435..2456
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2610..2629)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2729..2751
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2789..2807)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2875..2979
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2980..3048,3049..3840)
                     /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(3198..3217)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4011..4599
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4500..4519
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4661..4790
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 gccaggaaag ccgatgcttt ctgccaggca tagagctcgg gctgggcata aaagtcaggg
       61 cagagccatc tattgcttac atttgcttct gggatccaga tctttcgaag ctagcgctac
      121 cggtcgccac catggtctcc aagggcgagg agctgttcac cggggtggtg cccatcctgg
      181 tcgagctgga cggcgatgtc aacggccaca agttcagcgt gtccggcgag ggcgagggcg
      241 atgccaccta cggcaagctg accctgaagc tgatctgcac caccggcaag ctgcccgtgc
      301 cctggcccac cctcgtgacc accctgggct acggcgtgca gtgcttcgcc cgctaccccg
      361 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
      421 gcaccatctt cttcaaagac gacggcaact acaagacccg cgccgaggtg aagttcgagg
      481 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
      541 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc accgccgaca
      601 agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag gacggcggcg
      661 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
      721 ccgacaacca ctacctgagc taccagtcca agctgagcaa agaccccaac gagaagcgcg
      781 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
      841 tgtacaaggg ctctggtgct accaacttct cactgttgaa acaggcaggg gatgtagagg
      901 agaatccagg gcctggtgct agtggagact acaaagacca tgacggagat tataaagatc
      961 atgacatcga ttacaaggat gacgatgaca agtccggact cagatctgga ggctccggag
     1021 gcccagagcc agcgaagtct gctcccgccc cgaaaaaggg ctccaagaag gcggtgacta
     1081 aggcgcagaa gaaaggcggc aagaagcgca agcgcagccg caaggagagc tattccatct
     1141 atgtgtacaa ggttctgaag caggtccacc ctgacaccgg catttcgtcc aaggccatgg
     1201 gcatcatgaa ttcgtttgtg aacgacattt tcgagcgcat cgcaggagag gcttcccgcc
     1261 tggcgcatta caacaagcgc tcgaccatca cctcccggga gatccagacg gccgtgcgcc
     1321 tgctgctgcc tggggagttg gccaagcacg ccgtgtccga gggtactaag gccatcacca
     1381 agtacaccag cgctaagtaa tgaggcgcgc cgcggccgcg aattcgatat cataatcaac
     1441 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     1501 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     1561 tcattttctc ctccttgtat aaatcctggt tagttcttgc cacggcggaa ctcatcgccg
     1621 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggctc
     1681 gagagatctt cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     1741 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     1801 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     1861 aagggggagg attgggaaga caatagcagg catgagatct cacgtgcgga ccgagcggcc
     1921 gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
     1981 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga
     2041 gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg
     2101 cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta
     2161 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg
     2221 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     2281 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     2341 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     2401 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     2461 acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc
     2521 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     2581 acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     2641 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     2701 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     2761 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     2821 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     2881 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     2941 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     3001 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     3061 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     3121 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     3181 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     3241 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     3301 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     3361 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     3421 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     3481 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     3541 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     3601 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     3661 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     3721 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     3781 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     3841 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     3901 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     3961 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     4021 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     4081 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     4141 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     4201 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     4261 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     4321 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     4381 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     4441 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     4501 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     4561 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     4621 ttacggttcc tggccttttg ctggcctttt gctcacatgt cctgcaggca gctgcgcgct
     4681 cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag
     4741 cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct gcggccgcac
     4801 gcgttccaat gcttagagtt ttccctctaa agaggacctc ctgagccact tctgaacagg
     4861 acttggaaca aaggaatctt ggcgctttga tgatcgtgtc tgcagcagtt gttgaaatga
     4921 gtctgacact cgccagtgcc ttagtaggaa ttataattac atcaggagac cccaggcaac
     4981 caagtaaact gattagctac tgctccgagt tgatagattt ttttctctct aggtcataaa
     5041 tagggctgtg ggagatggtg gaagcatgct ggtgggaaca ggtcgagatg ggctgtacat
     5101 tcaggaagcc caaccaagtg gtgtgggcag ctccccctgc cccaccctgc caccggaaga
     5161 gacagtctga caggtgccca aggtagagtt cagttccctc ggccagggac tgcactcctg
     5221 atcttgcgtg ctgcagagag ctgtcctcag agcaggctga gggtggctga actgctaggc
     5281 gatgctctgg gaagccctcc
//
