LOCUS       sequence-473521-        4514 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0509h 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 4514)
  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 4514)
  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: AiP12469 - pAAV-AiE0509h-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4514
                     /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      526..535
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(532..534,535..537,538..1248)
                     /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(577..598)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(838..857)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1185..1206
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1341..1361)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1548..1659
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1777..1917
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1777..1906
                     /label=AAV2 ITR
     rep_origin      1992..2447
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2079..2098)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2289..2310
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2464..2483)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2583..2605
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2643..2661)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2729..2833
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2834..2902,2903..3694)
                     /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(3052..3071)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3865..4453
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4354..4373
                     /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 gcgtagagct acataccagc atgctaaaat ggcccatggc
      181 acaaacaaca tcgttagatg tcaggattct taccatccca ggctctcctt tgccttctcc
      241 tcttttcttc ttcctcctcc tcttccaaat gcatgcaaat aagggatgca cacttcatca
      301 aacagcccta aacctgtcag agcagctgct agcagaagcc aagttcatgc ctggctttct
      361 cccgtctcct gtgccagccc ccaccgcacg ccctcccgat cctggtgatt tttaaagtga
      421 aaagcaggta gcgagctcgg gctgggcata aaagtcaggg cagagccatc tattgcttac
      481 atttgcttct gggatccaga tctttcgaag ctagcgctac cggtcgccac catggtgagc
      541 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
      601 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
      661 accctgaagc tgatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
      721 accctgggct acggcgtgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac
      781 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
      841 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
      901 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
      961 tacaactaca acagccacaa cgtctatatc accgccgaca agcagaagaa cggcatcaag
     1021 gccaacttca agatccgcca caacatcgag gacggcggcg tgcagctcgc cgaccactac
     1081 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     1141 taccagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     1201 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta agtcgacggc
     1261 gcgccgcggc cgcgaattcg atatcataat caacctctgg attacaaaat ttgtgaaaga
     1321 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     1381 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     1441 tggttagttc ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     1501 gctcggctgt tgggcactga caattccgtg gctcgagaga tcttcgactg tgccttctag
     1561 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     1621 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     1681 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     1741 caggcatgag atctcacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg
     1801 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
     1861 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc
     1921 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa
     1981 gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     2041 cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc
     2101 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     2161 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
     2221 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     2281 ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc
     2341 ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta
     2401 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct
     2461 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     2521 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     2581 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     2641 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     2701 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     2761 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     2821 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     2881 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     2941 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     3001 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     3061 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     3121 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     3181 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     3241 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     3301 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     3361 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     3421 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     3481 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     3541 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     3601 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     3661 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     3721 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     3781 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     3841 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     3901 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     3961 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     4021 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     4081 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     4141 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     4201 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     4261 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     4321 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     4381 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     4441 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     4501 ttttgctcac atgt
//
