LOCUS       sequence-470493-        4635 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0562h 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 4635)
  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 4635)
  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: AiP12678 - pAAV-AiE0562h-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4635
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   180..309
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(371..959)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(451..470)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1130..1921,1922..1990))
                     /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     1753..1772
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1991..2095)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2163..2181
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2219..2241)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2341..2360
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2377..2832)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2514..2535)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2726..2745
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2907..3047
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2918..3047
                     /label=AAV2 ITR
     polyA_signal    3165..3276
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3463..3483
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3576..4286,4287..4289,4290..4292))
                     /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(3618..3639)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3967..3986
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4226..4247
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4289..4298
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 gcaactttgc ccccaccttc tctgcttcca ggaagtggct gagggctttg ccttccagga
       61 acagagaact gcccaagtgg gggttggtag ctgtgagtgt gctcataagt atgaaatatg
      121 tctctaggct aatggcttct tcctaaagtc cctgcctcca ggaaaacgcg tgcggccgca
      181 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
      241 cgggcgacca aaggtcgccc gacgcccggg cggcctcagt gagcgagcga gcgcgcagct
      301 gcctgcagga catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
      361 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
      421 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
      481 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
      541 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
      601 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
      661 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
      721 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
      781 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
      841 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
      901 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
      961 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     1021 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     1081 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     1141 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     1201 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     1261 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     1321 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     1381 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     1441 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     1501 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     1561 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     1621 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     1681 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     1741 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     1801 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     1861 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     1921 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     1981 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     2041 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     2101 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     2161 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc
     2221 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     2281 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
     2341 cggcatcaga gcagattgta ctgagagtgc accataaaat tgtaaacgtt aatattttgt
     2401 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag accgaaatcg
     2461 gcaaaatccc ttataaatca aaagaatagc ccgagataga gttgagtgtt gttccagttt
     2521 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     2581 atcagggcga tggcccacta cgtgaaccat cacccaaatc aagttttttg gggtcgaggt
     2641 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     2701 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctaaggcgc
     2761 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     2821 tacagggcgc gtactatggt tgctttgacg tatgcggtgt gaaataccgc acagatgcgt
     2881 aaggagaaaa taccgcatca ggcgcccctg caggcagctg cgcgctcgct cgctcactga
     2941 ggccgcccgg gcaaagcccg ggcgtcgggc gacctttggt cgcccggcct cagtgagcga
     3001 gcgagcgcgc agagagggag tggccaactc catcactagg ggttcctgcg gccgctcggt
     3061 ccgcacgtga gatctcatgc ctgctattgt cttcccaatc ctcccccttg ctgtcctgcc
     3121 ccaccccacc ccccagaata gaatgacacc tactcagaca atgcgatgca atttcctcat
     3181 tttattagga aaggacagtg ggagtggcac cttccagggt caaggaaggc acgggggagg
     3241 ggcaaacaac agatggctgg caactagaag gcacagtcga agatctctcg agccacggaa
     3301 ttgtcagtgc ccaacagccg agcccctgtc cagcagcggg caaggcaggc ggcgatgagt
     3361 tccgccgtgg caagaactaa ccaggattta tacaaggagg agaaaatgaa agccatacgg
     3421 gaagcaatag catgatacaa aggcattaaa gcagcgtatc cacatagcgt aaaaggagca
     3481 acatagttaa gaataccagt caatctttca caaattttgt aatccagagg ttgattatga
     3541 tatcgaattc gcggccgcgg cgcgccgtcg acttacttgt acagctcgtc catgccgaga
     3601 gtgatcccgg cggcggtcac gaactccagc aggaccatgt gatcgcgctt ctcgttgggg
     3661 tctttgctca gcttggactg gtagctcagg tagtggttgt cgggcagcag cacggggccg
     3721 tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct gcacgccgcc gtcctcgatg
     3781 ttgtggcgga tcttgaagtt ggccttgatg ccgttcttct gcttgtcggc ggtgatatag
     3841 acgttgtggc tgttgtagtt gtactccagc ttgtgcccca ggatgttgcc gtcctccttg
     3901 aagtcgatgc ccttcagctc gatgcggttc accagggtgt cgccctcgaa cttcacctcg
     3961 gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg tgcgctcctg gacgtagcct
     4021 tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta gcgggcgaag
     4081 cactgcacgc cgtagcccag ggtggtcacg agggtgggcc agggcacggg cagcttgccg
     4141 gtggtgcaga tcagcttcag ggtcagcttg ccgtaggtgg catcgccctc gccctcgccg
     4201 gacacgctga acttgtggcc gtttacgtcg ccgtccagct cgaccaggat gggcaccacc
     4261 ccggtgaaca gctcctcgcc cttgctcacc atggtggcga ccggtagcgc tagcttcgaa
     4321 agatctggat cccagaagca aatgtaagca atagatggct ctgccctgac ttttatgccc
     4381 agcccgagct ctaagtgggg gcaaatgtgg gattcaggcc ccagcctctc ctccacccca
     4441 gctgtctctg ggtgcaggag gctgtgggga tggctgtagg ggaagggtgg gcaacagtga
     4501 cagcctacct gaagaactgg gccccagcac cctccgtgtt tctgtgttat tctcccagcc
     4561 tccgggctgc tgcaagtccg gccctgactc accaagatgc ctcccccaac ccgggaacca
     4621 gggagaccag gcctt
//
