LOCUS       sequence-470864-        5024 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0082h_3xC3 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 5024)
  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 5024)
  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: AiP14484 -
            pAAV-AiE0082h_3xC3-minBG-SYFP2-WPRE...
FEATURES             Location/Qualifiers
     source          1..5024
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   651..780
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1686..1695
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1692..1694,1695..1697,1698..2408)
                     /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(1737..1758)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1998..2017)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2345..2366
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2501..2521)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2708..2819
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2937..3077
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2937..3066
                     /label=AAV2 ITR
     rep_origin      3152..3607
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3239..3258)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3449..3470
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3624..3643)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3743..3765
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3803..3821)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3889..3993
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3994..4062,4063..4854)
                     /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(4212..4231)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt cctgcaggca
      661 gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt ggtcgcccgg
      721 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
      781 gcggccgcac gcgtccatct ccctctctgc ctcccgtttc ccctttactc tttactgtta
      841 gttggcctgg ggttcctggg ctgaagtgga gtttcctcct gtctccgtgt gtttgctgct
      901 ctggtttgct cttcttccag ggccccttgg taaaagaaga gctgctgaca gggtagagtg
      961 gaggtgggga agtgaccacc agactgggag tgatgtaggt tgtggtcatc aatgatgtag
     1021 gttgctgtca ccaggacact caaatcagct gtgaccagag ccatctccct ctctgcctcc
     1081 cgtttcccct ttactcttta ctgttagttg gcctggggtt cctgggctga agtggagttt
     1141 cctcctgtct ccgtgtgttt gctgctctgg tttgctcttc ttccagggcc ccttggtaaa
     1201 agaagagctg ctgacagggt agagtggagg tggggaagtg accaccagac tgggagtgat
     1261 gtaggttgtg gtcatcaatg atgtaggttg ctgtcaccag gacactcaaa tcagctgtga
     1321 ccagagccat ctccctctct gcctcccgtt tcccctttac tctttactgt tagttggcct
     1381 ggggttcctg ggctgaagtg gagtttcctc ctgtctccgt gtgtttgctg ctctggtttg
     1441 ctcttcttcc agggcccctt ggtaaaagaa gagctgctga cagggtagag tggaggtggg
     1501 gaagtgacca ccagactggg agtgatgtag gttgtggtca tcaatgatgt aggttgctgt
     1561 caccaggaca ctcaaatcag ctgtgaccag aggagctcgg gctgggcata aaagtcaggg
     1621 cagagccatc tattgcttac atttgcttct gggatccaga tctttcgaag ctagcgctac
     1681 cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     1741 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     1801 atgccaccta cggcaagctg accctgaagc tgatctgcac caccggcaag ctgcccgtgc
     1861 cctggcccac cctcgtgacc accctgggct acggcgtgca gtgcttcgcc cgctaccccg
     1921 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     1981 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     2041 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     2101 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc accgccgaca
     2161 agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag gacggcggcg
     2221 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     2281 ccgacaacca ctacctgagc taccagtcca agctgagcaa agaccccaac gagaagcgcg
     2341 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     2401 tgtacaagta agtcgacggc gcgccgcggc cgcgaattcg atatcataat caacctctgg
     2461 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     2521 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     2581 tctcctcctt gtataaatcc tggttagttc ttgccacggc ggaactcatc gccgcctgcc
     2641 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gctcgagaga
     2701 tcttcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     2761 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     2821 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     2881 gaggattggg aagacaatag caggcatgag atctcacgtg cggaccgagc ggccgcagga
     2941 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     3001 gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc
     3061 gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     3121 ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg
     3181 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct
     3241 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3301 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     3361 cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     3421 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     3481 aactctatct cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg
     3541 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt
     3601 acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     3661 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     3721 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     3781 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     3841 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     3901 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     3961 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     4021 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     4081 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     4141 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     4201 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     4261 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     4321 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     4381 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     4441 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     4501 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     4561 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     4621 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     4681 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     4741 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     4801 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     4861 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     4921 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     4981 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttc
//
