LOCUS       sequence-450544-        5493 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0078h_3xC2 is an enhancer sequence, designed to drive 
            AAV-mediated transgene expression in forebrain glutamatergic 
            neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5493)
  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 5493)
  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: AiP11954 -
            pAAV-hsA2-AiE0078h_3xC2-minRho-SYFP...
FEATURES             Location/Qualifiers
     source          1..5493
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1530..1539
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1536..1538,1539..1541,1542..2252)
                     /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(1581..1602)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1842..1861)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2189..2210
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2320..2340)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2527..2638
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2756..2896
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2756..2885
                     /label=AAV2 ITR
     rep_origin      2971..3426
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3058..3077)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3268..3289
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3443..3462)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3562..3584
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3622..3640)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3708..3812
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3813..3881,3882..4673)
                     /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(4031..4050)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4844..5432
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5333..5352
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcaa cgcgtttaga acaatggctg gcccatagta aatgccgtgt
      181 tagtgtgtta gttgctgttc ttccacgtca gaagaggcac agacaaatta ccaccaggtg
      241 gcgctcagag tctgcggagg catcacaaca gccctgaatt tgaatcctgc tctgccactg
      301 cctagttgag accttttact acctgactag ctgtttgtgt attttaggtg tttgtttgaa
      361 aagatccggg ttcacactaa tcaggcccaa cggaaggcca tattagcaat ttggcaggta
      421 cccgagggcc atacctaatc tgcataaaat gaagcagatt gcaaccgccc tcatcttttt
      481 tatttttaaa ctggtttttg aagcagagca taaaatctca gagggagaga cagaagatgc
      541 tagtgcatac attttccttc atgcctttat tttcattctt tttgcacaaa ccatcttcct
      601 gaatggctgt ttacctaaag aagaataaca aaataaaagg tgctaggaaa tggagtaggc
      661 agagatcgag cagagccctc atcacacaga ctgaaaagat ccgggttcac actaatcagg
      721 cccaacggaa ggccatatta gcaatttggc aggtacccga gggccatacc taatctgcat
      781 aaaatgaagc agattgcaac cgccctcatc ttttttattt ttaaactggt ttttgaagca
      841 gagcataaaa tctcagaggg agagacagaa gatgctagtg catacatttt ccttcatgcc
      901 tttattttca ttctttttgc acaaaccatc ttcctgaatg gctgtttacc taaagaagaa
      961 taacaaaata aaaggtgcta ggaaatggag taggcagaga tctgggtgag taagagtagg
     1021 tggcaacgaa aagatccggg ttcacactaa tcaggcccaa cggaaggcca tattagcaat
     1081 ttggcaggta cccgagggcc atacctaatc tgcataaaat gaagcagatt gcaaccgccc
     1141 tcatcttttt tatttttaaa ctggtttttg aagcagagca taaaatctca gagggagaga
     1201 cagaagatgc tagtgcatac attttccttc atgcctttat tttcattctt tttgcacaaa
     1261 ccatcttcct gaatggctgt ttacctaaag aagaataaca aaataaaagg tgctaggaaa
     1321 tggagtaggc agagatcgaa ttcgatatca taatcaacca taggtaccga gctcgggatt
     1381 cagccgggag cttagggagg ggaggtcact tcataagggc ctgggggggg agttggagcc
     1441 acgagtcgtc cagccggagc cccgtgtggc tgagctccgg cctcagaagc atccccgggt
     1501 tggatccttc gaagctagcg ctaccggtcg ccaccatggt gagcaagggc gaggagctgt
     1561 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     1621 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagctgatct
     1681 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg ggctacggcg
     1741 tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     1801 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     1861 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     1921 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     1981 acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc
     2041 gccacaacat cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag aacaccccca
     2101 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag tccaagctga
     2161 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     2221 ggatcactct cggcatggac gagctgtaca agtaagtcga catcataatc aacctctgga
     2281 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     2341 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     2401 ctcctccttg tataaatcct ggttagttct tgccacggcg gaactcatcg ccgcctgcct
     2461 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg ctcgagagat
     2521 cttcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     2581 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     2641 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     2701 aggattggga agacaatagc aggcatgaga tctcacgtgc ggaccgagcg gccgcaggaa
     2761 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg
     2821 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg
     2881 cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     2941 tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg
     3001 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc
     3061 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3121 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     3181 ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     3241 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3301 actctatctc gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt
     3361 taaaaaatga gctgatttaa caaaaattta acgggaattt taacaaaata ttaacgttta
     3421 caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     3481 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     3541 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     3601 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     3661 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     3721 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     3781 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     3841 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     3901 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     3961 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     4021 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     4081 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     4141 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     4201 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     4261 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     4321 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     4381 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     4441 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     4501 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     4561 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     4621 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     4681 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     4741 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     4801 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     4861 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4921 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     4981 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     5041 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     5101 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     5161 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     5221 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     5281 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     5341 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     5401 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     5461 tcctggcctt ttgctggcct tttgctcaca tgt
//
