LOCUS       sequence-470443-        4650 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0397h 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 4650)
  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 4650)
  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: AiP12159 - pAAV-AiE0397h-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4650
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(120..139)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     239..261
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(299..317)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        385..489
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(490..558,559..1350)
                     /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(708..727)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1521..2109
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2010..2029
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2171..2300
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      2832..2841
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2838..2840,2841..2843,2844..3554)
                     /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(2883..2904)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3144..3163)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3491..3512
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3647..3667)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3854..3965
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4083..4223
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4083..4212
                     /label=AAV2 ITR
     rep_origin      join(4298..4650,1..103)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4385..4404)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4595..4616
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa
       61 aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt
      121 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
      181 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
      241 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
      301 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
      361 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
      421 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
      481 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
      541 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
      601 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
      661 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
      721 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
      781 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
      841 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
      901 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
      961 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     1021 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     1081 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     1141 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     1201 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     1261 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     1321 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     1381 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     1441 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     1501 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     1561 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     1621 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     1681 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     1741 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     1801 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     1861 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     1921 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     1981 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     2041 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     2101 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     2161 gctcacatgt cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg
     2221 ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa
     2281 ctccatcact aggggttcct gcggccgcac gcgtggagat ttggtcctga ttttgaatgg
     2341 tttttcttgg tgggcagtgg ttgtctgagg ctttgaactg atctttaaca cacaggaatg
     2401 agttctaggc acttctggag tggtccagag cacctctaaa tacgaagcag cagatgaggt
     2461 catggtaatt cggggagtca gaagtggaag ggtggggggt tgaaagctat aatcactgtc
     2521 aaaggggtgt tttctgtaga caagtgaatg gagctattgt acacaaactc agtggttgcc
     2581 aagcatgtgg cccaattatt ttgccttcat gagcatggca cggaaggatc tctttagaga
     2641 acttgattca cacaagtttg tgtttagtct aacggtagca ggcaaaagcc tggggaattt
     2701 gaacctaaga gtctttacaa gttcctgcct cccaaatcga gctcgggctg ggcataaaag
     2761 tcagggcaga gccatctatt gcttacattt gcttctggga tccagatctt tcgaagctag
     2821 cgctaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     2881 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     2941 agggcgatgc cacctacggc aagctgaccc tgaagctgat ctgcaccacc ggcaagctgc
     3001 ccgtgccctg gcccaccctc gtgaccaccc tgggctacgg cgtgcagtgc ttcgcccgct
     3061 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     3121 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     3181 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     3241 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcaccg
     3301 ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac atcgaggacg
     3361 gcggcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     3421 tgctgcccga caaccactac ctgagctacc agtccaagct gagcaaagac cccaacgaga
     3481 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     3541 acgagctgta caagtaagtc gacggcgcgc cgcggccgcg aattcgatat cataatcaac
     3601 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     3661 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     3721 tcattttctc ctccttgtat aaatcctggt tagttcttgc cacggcggaa ctcatcgccg
     3781 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggctc
     3841 gagagatctt cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     3901 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     3961 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     4021 aagggggagg attgggaaga caatagcagg catgagatct cacgtgcgga ccgagcggcc
     4081 gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
     4141 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga
     4201 gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg
     4261 cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta
     4321 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg
     4381 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     4441 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     4501 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     4561 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     4621 acactcaact ctatctcggg ctattctttt
//
