LOCUS       sequence-470691-        5630 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE1015h 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 5630)
  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 5630)
  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: AiP13471 -
            pAAV-AiE1015h-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5630
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     143..162
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   324..464
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   335..464
                     /label=AAV2 ITR
     polyA_signal    582..693
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     880..900
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(987..1364)
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     CDS             complement(1395..1460)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(1473..1529)
                     /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             complement(join(1539..2249,2250..2252,2253..2255))
                     /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(1581..1602)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1930..1949
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     regulatory      2252..2261
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   3227..3356
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3418..4006)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3498..3517)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4177..4968,4969..5037))
                     /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     4800..4819
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5038..5142)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5210..5228
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5266..5288)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5388..5407
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(join(5424..5630,1..249))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5561..5582)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 agggcgatgg cccactacgt gaaccatcac ccaaatcaag ttttttgggg tcgaggtgcc
       61 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
      121 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct aaggcgctgg
      181 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
      241 agggcgcgta ctatggttgc tttgacgtat gcggtgtgaa ataccgcaca gatgcgtaag
      301 gagaaaatac cgcatcaggc gcccctgcag gcagctgcgc gctcgctcgc tcactgaggc
      361 cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg
      421 agcgcgcaga gagggagtgg ccaactccat cactaggggt tcctgcggcc gctcggtccg
      481 cacgtgagat ctcatgcctg ctattgtctt cccaatcctc ccccttgctg tcctgcccca
      541 ccccaccccc cagaatagaa tgacacctac tcagacaatg cgatgcaatt tcctcatttt
      601 attaggaaag gacagtggga gtggcacctt ccagggtcaa ggaaggcacg ggggaggggc
      661 aaacaacaga tggctggcaa ctagaaggca cagtcgaaga tctctcgagc cacggaattg
      721 tcagtgccca acagccgagc ccctgtccag cagcgggcaa ggcaggcggc gatgagttcc
      781 gccgtggcaa gaactaacca ggatttatac aaggaggaga aaatgaaagc catacgggaa
      841 gcaatagcat gatacaaagg cattaaagca gcgtatccac atagcgtaaa aggagcaaca
      901 tagttaagaa taccagtcaa tctttcacaa attttgtaat ccagaggttg attatgatat
      961 cgaattcgcg gccgcggcgc gcctcattac ttagcgctgg tgtacttggt gatggcctta
     1021 gtaccctcgg acacggcgtg cttggccaac tccccaggca gcagcaggcg cacggccgtc
     1081 tggatctccc gggaggtgat ggtcgagcgc ttgttgtaat gcgccaggcg ggaagcctct
     1141 cctgcgatgc gctcgaaaat gtcgttcaca aacgaattca tgatgcccat ggccttggac
     1201 gaaatgccgg tgtcagggtg gacctgcttc agaaccttgt acacatagat ggaatagctc
     1261 tccttgcggc tgcgcttgcg cttcttgccg cctttcttct gcgccttagt caccgccttc
     1321 ttggagccct ttttcggggc gggagcagac ttcgctggct ctgggcctcc ggagcctcca
     1381 gatctgagtc cggacttgtc atcgtcatcc ttgtaatcga tgtcatgatc tttataatct
     1441 ccgtcatggt ctttgtagtc tccactagca ccaggccctg gattctcctc tacatcccct
     1501 gcctgtttca acagtgagaa gttggtagca ccagagccct tgtacagctc gtccatgccg
     1561 agagtgatcc cggcggcggt cacgaactcc agcaggacca tgtgatcgcg cttctcgttg
     1621 gggtctttgc tcagcttgga ctggtagctc aggtagtggt tgtcgggcag cagcacgggg
     1681 ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga gctgcacgcc gccgtcctcg
     1741 atgttgtggc ggatcttgaa gttggccttg atgccgttct tctgcttgtc ggcggtgata
     1801 tagacgttgt ggctgttgta gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc
     1861 ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc
     1921 tcggcgcggg tcttgtagtt gccgtcgtct ttgaagaaga tggtgcgctc ctggacgtag
     1981 ccttcgggca tggcggactt gaagaagtcg tgctgcttca tgtggtcggg gtagcgggcg
     2041 aagcactgca cgccgtagcc cagggtggtc acgagggtgg gccagggcac gggcagcttg
     2101 ccggtggtgc agatcagctt cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg
     2161 ccggacacgc tgaacttgtg gccgttgaca tcgccgtcca gctcgaccag gatgggcacc
     2221 accccggtga acagctcctc gcccttggag accatggtgg cgaccggtag cgctagcttc
     2281 gaaagatctg gatcccagaa gcaaatgtaa gcaatagatg gctctgccct gacttttatg
     2341 cccagcccga gctcttccca gcatagagag gatggaggcc agggatgctc ctcagcaccc
     2401 tgcggtactc agcacagccc cctaccacag ccattggccc caaatgtcag caccaggtgt
     2461 gggtgacaat atctaacccg ctgtgcaaag atcaggatgt cacggcatat tatcagttgt
     2521 cttgcttctt ccttcagacc acagcatcac ccttattttc tagaactttc catccataaa
     2581 actgcagccc tcctccagcc caggagcggc tgcaccctta gcaggcattc attgagtgct
     2641 gcctggatgc cgacagcctt ggtagggaat acaccatgct gccgaaattg gcctcgggtg
     2701 ttccgggggc tccctccagc agcccctcca tctccctggg gttggggtcc ttggtggccg
     2761 tcaccgagcc agggaaacag gagacaagac caaccgccca gccactggtt cccccacccc
     2821 tcccattctt ttcataagcg caaatttccc ctgccaattt gcatatcccg tttttagcgt
     2881 ctttcttttc taggttttac ataatcatct cattattaga tgctggcaca tgaacttaat
     2941 gttccctcaa tcacatttta cagcattttc tagcgtaagt agctaattgc ctgtcgtcct
     3001 tctcttctct ctgtctcttt tttttttttg ataaactttt cattttagaa tagtttcaga
     3061 tttacagaaa aactgggaat gcagtgcaga cagtttcatc tcctcaccac ccaattttcc
     3121 ttagtattca catccaatgg catggtgcgt ttgtcacagc gaataaacca gtattgatac
     3181 atcttcatta acccaagtct gcactctgtt caacgcgtgc ggccgcagga acccctagtg
     3241 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag
     3301 gtcgcccgac gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggacat
     3361 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     3421 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     3481 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     3541 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3601 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     3661 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3721 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     3781 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     3841 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     3901 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     3961 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     4021 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     4081 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     4141 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     4201 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     4261 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     4321 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     4381 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     4441 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     4501 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     4561 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     4621 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     4681 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     4741 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     4801 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     4861 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     4921 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     4981 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     5041 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     5101 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     5161 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     5221 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     5281 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     5341 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     5401 gattgtactg agagtgcacc ataaaattgt aaacgttaat attttgttaa aattcgcgtt
     5461 aaatttttgt taaatcagct cattttttaa ccaatagacc gaaatcggca aaatccctta
     5521 taaatcaaaa gaatagcccg agatagagtt gagtgttgtt ccagtttgga acaagagtcc
     5581 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
//
