LOCUS       sequence-470446-        4490 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0407h 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 4490)
  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 4490)
  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: AiP12167 - pAAV-AiE0407h-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4490
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(170..758)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(250..269)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(929..1720,1721..1789))
                     /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     1552..1571
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1790..1894)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1962..1980
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2018..2040)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2140..2159
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2176..2631)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2313..2334)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2525..2544
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2706..2846
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2717..2846
                     /label=AAV2 ITR
     polyA_signal    2964..3075
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3262..3282
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3375..4085,4086..4088,4089..4091))
                     /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(3417..3438)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3766..3785
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4025..4046
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4088..4097
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   join(4469..4490,1..108)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
       61 acgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcaggac atgtgagcaa
      121 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
      181 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
      241 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
      301 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
      361 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
      421 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
      481 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
      541 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
      601 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
      661 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
      721 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
      781 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
      841 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
      901 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
      961 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     1021 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     1081 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     1141 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     1201 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     1261 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     1321 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     1381 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     1441 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     1501 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     1561 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     1621 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     1681 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     1741 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     1801 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     1861 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     1921 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     1981 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     2041 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     2101 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     2161 tgagagtgca ccataaaatt gtaaacgtta atattttgtt aaaattcgcg ttaaattttt
     2221 gttaaatcag ctcatttttt aaccaataga ccgaaatcgg caaaatccct tataaatcaa
     2281 aagaatagcc cgagatagag ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     2341 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     2401 gtgaaccatc acccaaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     2461 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     2521 aggaagggaa gaaagcgaaa ggagcgggcg ctaaggcgct ggcaagtgta gcggtcacgc
     2581 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tactatggtt
     2641 gctttgacgt atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     2701 gcgcccctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg caaagcccgg
     2761 gcgtcgggcg acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt
     2821 ggccaactcc atcactaggg gttcctgcgg ccgctcggtc cgcacgtgag atctcatgcc
     2881 tgctattgtc ttcccaatcc tcccccttgc tgtcctgccc caccccaccc cccagaatag
     2941 aatgacacct actcagacaa tgcgatgcaa tttcctcatt ttattaggaa aggacagtgg
     3001 gagtggcacc ttccagggtc aaggaaggca cgggggaggg gcaaacaaca gatggctggc
     3061 aactagaagg cacagtcgaa gatctctcga gccacggaat tgtcagtgcc caacagccga
     3121 gcccctgtcc agcagcgggc aaggcaggcg gcgatgagtt ccgccgtggc aagaactaac
     3181 caggatttat acaaggagga gaaaatgaaa gccatacggg aagcaatagc atgatacaaa
     3241 ggcattaaag cagcgtatcc acatagcgta aaaggagcaa catagttaag aataccagtc
     3301 aatctttcac aaattttgta atccagaggt tgattatgat atcgaattcg cggccgcggc
     3361 gcgccgtcga cttacttgta cagctcgtcc atgccgagag tgatcccggc ggcggtcacg
     3421 aactccagca ggaccatgtg atcgcgcttc tcgttggggt ctttgctcag cttggactgg
     3481 tagctcaggt agtggttgtc gggcagcagc acggggccgt cgccgatggg ggtgttctgc
     3541 tggtagtggt cggcgagctg cacgccgccg tcctcgatgt tgtggcggat cttgaagttg
     3601 gccttgatgc cgttcttctg cttgtcggcg gtgatataga cgttgtggct gttgtagttg
     3661 tactccagct tgtgccccag gatgttgccg tcctccttga agtcgatgcc cttcagctcg
     3721 atgcggttca ccagggtgtc gccctcgaac ttcacctcgg cgcgggtctt gtagttgccg
     3781 tcgtccttga agaagatggt gcgctcctgg acgtagcctt cgggcatggc ggacttgaag
     3841 aagtcgtgct gcttcatgtg gtcggggtag cgggcgaagc actgcacgcc gtagcccagg
     3901 gtggtcacga gggtgggcca gggcacgggc agcttgccgg tggtgcagat cagcttcagg
     3961 gtcagcttgc cgtaggtggc atcgccctcg ccctcgccgg acacgctgaa cttgtggccg
     4021 tttacgtcgc cgtccagctc gaccaggatg ggcaccaccc cggtgaacag ctcctcgccc
     4081 ttgctcacca tggtggcgac cggtagcgct agcttcgaaa gatctggatc ccagaagcaa
     4141 atgtaagcaa tagatggctc tgccctgact tttatgccca gcccgagctc ctgtgctccc
     4201 ttctaccccc acccccgcag ccagtgctgg ttccccacac ctggtcacta ggctctgtaa
     4261 gcttctcagg gccagccagt tctgcgctgg caccaagcca gtctcagcaa acaaagaaca
     4321 atcggcatgt tgttctcaat ccagttcaca cttaaaaccc tctgggccag gcacaacctc
     4381 cctccactgc cctcagactc tttgactttc tgattgactg ggtggtgaca cctgggctgg
     4441 ggagcaaagc agtaacgcgt gcggccgcag gaacccctag tgatggagtt
//
