LOCUS       sequence-452702-        4540 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0374h 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 4540)
  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 4540)
  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: AiP12493 - pAAV-AiE0374h-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4540
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     regulatory      552..561
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(558..560,561..563,564..1274)
                     /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(603..624)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(864..883)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1211..1232
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1367..1387)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1574..1685
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1803..1943
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1803..1932
                     /label=AAV2 ITR
     rep_origin      2018..2473
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2105..2124)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2315..2336
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2490..2509)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2609..2631
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2669..2687)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2755..2859
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2860..2928,2929..3720)
                     /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(3078..3097)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3891..4479
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4380..4399
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgttgagg agaggaggcc gagaaactgg
      181 gctgagcagg acaaaacctc ccctgtgaac cctgccaagg ggaggggccc cttattaaaa
      241 cttactgggg catagccagg cccccctgga gggagggggt ctgaggggca gggaaggggg
      301 ctgggtggag gcaactagga ctctggcctc cagccagtcc tcccctgggt ggctgtttct
      361 gtggctgggc tgtggttcac accatctgta gcccttcagt tgtccccttc tccataccaa
      421 agtgctgttt cgcctagtcc tgccctggtc ccattcatga gctcgggctg ggcataaaag
      481 tcagggcaga gccatctatt gcttacattt gcttctggga tccagatctt tcgaagctag
      541 cgctaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
      601 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
      661 agggcgatgc cacctacggc aagctgaccc tgaagctgat ctgcaccacc ggcaagctgc
      721 ccgtgccctg gcccaccctc gtgaccaccc tgggctacgg cgtgcagtgc ttcgcccgct
      781 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
      841 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
      901 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
      961 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcaccg
     1021 ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac atcgaggacg
     1081 gcggcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     1141 tgctgcccga caaccactac ctgagctacc agtccaagct gagcaaagac cccaacgaga
     1201 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     1261 acgagctgta caagtaagtc gacggcgcgc cgcggccgcg aattcgatat cataatcaac
     1321 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     1381 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     1441 tcattttctc ctccttgtat aaatcctggt tagttcttgc cacggcggaa ctcatcgccg
     1501 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggctc
     1561 gagagatctt cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     1621 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     1681 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     1741 aagggggagg attgggaaga caatagcagg catgagatct cacgtgcgga ccgagcggcc
     1801 gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
     1861 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga
     1921 gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg
     1981 cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta
     2041 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     2101 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     2161 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     2221 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     2281 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     2341 acactcaacc ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggcc
     2401 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     2461 acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     2521 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     2581 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     2641 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     2701 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     2761 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     2821 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     2881 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     2941 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     3001 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     3061 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     3121 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     3181 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     3241 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     3301 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     3361 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     3421 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     3481 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     3541 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     3601 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     3661 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     3721 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     3781 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     3841 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     3901 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     3961 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     4021 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     4081 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     4141 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     4201 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     4261 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     4321 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     4381 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     4441 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     4501 ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
