LOCUS       sequence-470504-        4827 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0692h 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 4827)
  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 4827)
  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: AiP12690 - pAAV-AiE0692h-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4827
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     15..34
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   196..336
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   207..336
                     /label=AAV2 ITR
     polyA_signal    454..565
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     752..772
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(865..1575,1576..1578,1579..1581))
                     /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(907..928)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1256..1275
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1515..1536
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1578..1587
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   2296..2425
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2487..3075)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2567..2586)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3246..4037,4038..4106))
                     /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     3869..3888
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4107..4211)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4279..4297
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4335..4357)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4457..4476
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(join(4493..4827,1..121))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4630..4651)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctaaggcgct ggcaagtgta
       61 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
      121 tactatggtt gctttgacgt atgcggtgtg aaataccgca cagatgcgta aggagaaaat
      181 accgcatcag gcgcccctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg
      241 caaagcccgg gcgtcgggcg acctttggtc gcccggcctc agtgagcgag cgagcgcgca
      301 gagagggagt ggccaactcc atcactaggg gttcctgcgg ccgctcggtc cgcacgtgag
      361 atctcatgcc tgctattgtc ttcccaatcc tcccccttgc tgtcctgccc caccccaccc
      421 cccagaatag aatgacacct actcagacaa tgcgatgcaa tttcctcatt ttattaggaa
      481 aggacagtgg gagtggcacc ttccagggtc aaggaaggca cgggggaggg gcaaacaaca
      541 gatggctggc aactagaagg cacagtcgaa gatctctcga gccacggaat tgtcagtgcc
      601 caacagccga gcccctgtcc agcagcgggc aaggcaggcg gcgatgagtt ccgccgtggc
      661 aagaactaac caggatttat acaaggagga gaaaatgaaa gccatacggg aagcaatagc
      721 atgatacaaa ggcattaaag cagcgtatcc acatagcgta aaaggagcaa catagttaag
      781 aataccagtc aatctttcac aaattttgta atccagaggt tgattatgat atcgaattcg
      841 cggccgcggc gcgccgtcga cttacttgta cagctcgtcc atgccgagag tgatcccggc
      901 ggcggtcacg aactccagca ggaccatgtg atcgcgcttc tcgttggggt ctttgctcag
      961 cttggactgg tagctcaggt agtggttgtc gggcagcagc acggggccgt cgccgatggg
     1021 ggtgttctgc tggtagtggt cggcgagctg cacgccgccg tcctcgatgt tgtggcggat
     1081 cttgaagttg gccttgatgc cgttcttctg cttgtcggcg gtgatataga cgttgtggct
     1141 gttgtagttg tactccagct tgtgccccag gatgttgccg tcctccttga agtcgatgcc
     1201 cttcagctcg atgcggttca ccagggtgtc gccctcgaac ttcacctcgg cgcgggtctt
     1261 gtagttgccg tcgtccttga agaagatggt gcgctcctgg acgtagcctt cgggcatggc
     1321 ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag cgggcgaagc actgcacgcc
     1381 gtagcccagg gtggtcacga gggtgggcca gggcacgggc agcttgccgg tggtgcagat
     1441 cagcttcagg gtcagcttgc cgtaggtggc atcgccctcg ccctcgccgg acacgctgaa
     1501 cttgtggccg tttacgtcgc cgtccagctc gaccaggatg ggcaccaccc cggtgaacag
     1561 ctcctcgccc ttgctcacca tggtggcgac cggtagcgct agcttcgaaa gatctggatc
     1621 ccagaagcaa atgtaagcaa tagatggctc tgccctgact tttatgccca gcccgagctc
     1681 tcacatacct caccgcaaac gaaaatgacg ataacagtga cagctccatt gacaatgata
     1741 atgaccacca ctgaatcagc agggctctta tcctttctct tcctcccaaa tgtacagtgc
     1801 atcggaggat gctctccaat ctcactgcgc tgttgaaagg aataaatgat aacatgtcac
     1861 agcaacattt tgcaaatata aaatgctctc caaatggttt tactcccagg gcagtccctg
     1921 gctctgaagg aaagtggctg tctacccact ttagcaaaga cttgtgttct gggatacacc
     1981 cggagcaata ccttgcatct gctaaaaggt ttcagagcaa gaaaacatct tgcctgagca
     2041 gaaatatggg cagcttcaaa attgctggca gtggtaaagc tctgactgat gctgtctccc
     2101 tgaattcggt gtctggttga accttatcct ccccttttgt tctggacagg ttataaggac
     2161 atttttactg caagtcaata agaccttttg tctctgtggc cagggtgctg gtcttcctgt
     2221 gggctgcacc tgccaggagg ggctctgtgg cagcaattgc ggtctgcccc cagaacttca
     2281 gacgcgtgcg gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     2341 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcggc ctcagtgagc
     2401 gagcgagcgc gcagctgcct gcaggacatg tgagcaaaag gccagcaaaa ggccaggaac
     2461 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     2521 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     2581 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     2641 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     2701 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     2761 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     2821 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     2881 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     2941 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     3001 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     3061 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     3121 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     3181 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     3241 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     3301 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     3361 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     3421 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     3481 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     3541 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     3601 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     3661 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     3721 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     3781 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     3841 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     3901 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     3961 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     4021 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     4081 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     4141 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     4201 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     4261 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
     4321 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     4381 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     4441 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca taaaattgta
     4501 aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     4561 caatagaccg aaatcggcaa aatcccttat aaatcaaaag aatagcccga gatagagttg
     4621 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     4681 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc caaatcaagt
     4741 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     4801 agagcttgac ggggaaagcc ggcgaac
//
