LOCUS       sequence-470807-        5125 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0354m_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 5125)
  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 5125)
  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: AiP12566 -
            pAAV-3xSP10ins-AiE0354m_3xC3-minRho...
FEATURES             Location/Qualifiers
     source          1..5125
                     /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      1787..1796
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1793..1795,1796..1798,1799..2509)
                     /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(1838..1859)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2099..2118)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2446..2467
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2602..2622)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2809..2920
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3038..3178
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3038..3167
                     /label=AAV2 ITR
     rep_origin      3253..3708
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3340..3359)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3550..3571
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3725..3744)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3844..3866
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3904..3922)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3990..4094
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4095..4163,4164..4955)
                     /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(4313..4332)
                     /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 gcgccggtac cgaagctacc cctaacacac tattctacac acagaaaatg
      841 ctcttcacta ggaagctacc cctaacacac tattctacac acagaaaatg ctcttcacta
      901 ggaagctacc cctaacacac tattctacac acagaaaatg ctcttcacta gacgcgtgac
      961 ctttaacaat gaatctttaa atttaaccgg gtcccccaga gatcaccgca cacaaggttt
     1021 cattgtgttt cgatctgagg tgagattcaa atgtgggccg tgggattcct gttttgggta
     1081 gagcggttaa cacagaacag taggataggt cagcaattat cagccaagaa caaaggagaa
     1141 ccctgatgct cagtctacct tattatgggg tctcactttg gacctttaac aatgaatctt
     1201 taaatttaac cgggtccccc agagatcacc gcacacaagg tttcattgtg tttcgatctg
     1261 aggtgagatt caaatgtggg ccgtgggatt cctgttttgg gtagagcggt taacacagaa
     1321 cagtaggata ggtcagcaat tatcagccaa gaacaaagga gaaccctgat gctcagtcta
     1381 ccttattatg gggtctcact ttggaccttt aacaatgaat ctttaaattt aaccgggtcc
     1441 cccagagatc accgcacaca aggtttcatt gtgtttcgat ctgaggtgag attcaaatgt
     1501 gggccgtggg attcctgttt tgggtagagc ggttaacaca gaacagtagg ataggtcagc
     1561 aattatcagc caagaacaaa ggagaaccct gatgctcagt ctaccttatt atggggtctc
     1621 actttggagc tcgattcagc cgggagctta gggaggggag gtcacttcat aagggcttgg
     1681 ggggggagtt ggagccacga gtcgtccagc cggagccccg tgtggctgtg ctccggcctc
     1741 agaagcatcc ccggatccag atctttcgaa gctagcgcta ccggtcgcca ccatggtgag
     1801 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     1861 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     1921 gaccctgaag ctgatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     1981 caccctgggc tacggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga
     2041 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     2101 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     2161 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     2221 gtacaactac aacagccaca acgtctatat caccgccgac aagcagaaga acggcatcaa
     2281 ggccaacttc aagatccgcc acaacatcga ggacggcggc gtgcagctcg ccgaccacta
     2341 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     2401 ctaccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     2461 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt aagtcgacgg
     2521 cgcgccgcgg ccgcgaattc gatatcataa tcaacctctg gattacaaaa tttgtgaaag
     2581 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     2641 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     2701 ctggttagtt cttgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     2761 ggctcggctg ttgggcactg acaattccgt ggctcgagag atcttcgact gtgccttcta
     2821 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     2881 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     2941 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     3001 gcaggcatga gatctcacgt gcggaccgag cggccgcagg aacccctagt gatggagttg
     3061 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
     3121 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg
     3181 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa
     3241 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     3301 gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt
     3361 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     3421 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt
     3481 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     3541 tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt
     3601 cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt
     3661 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc
     3721 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     3781 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     3841 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     3901 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     3961 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     4021 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     4081 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     4141 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     4201 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     4261 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     4321 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     4381 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     4441 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     4501 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     4561 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     4621 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     4681 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     4741 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     4801 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     4861 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     4921 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     4981 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     5041 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     5101 ccgtagaaaa gatcaaagga tcttc
//
