LOCUS       sequence-450553-        5135 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0354m_3xC2 is an enhancer sequence, designed to drive 
            AAV-mediated transgene expression in cortical Vip+ neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5135)
  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 5135)
  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: AiP12565 -
            pAAV-3xSP10ins-AiE0354m_3xC2-minRho...
FEATURES             Location/Qualifiers
     source          1..5135
                     /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      1147..1156
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1153..1155,1156..1158,1159..1869)
                     /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(1198..1219)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1459..1478)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1806..1827
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1962..1982)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2169..2280
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2398..2538
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2398..2527
                     /label=AAV2 ITR
     rep_origin      2613..3068
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2700..2719)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2910..2931
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3085..3104)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3204..3226
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3264..3282)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3350..3454
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3455..3523,3524..4315)
                     /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(3673..3692)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4486..5074
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4975..4994
                     /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 gcggccgcac gcgccggtac cgaagctacc cctaacacac tattctacac
      181 acagaaaatg ctcttcacta ggaagctacc cctaacacac tattctacac acagaaaatg
      241 ctcttcacta ggaagctacc cctaacacac tattctacac acagaaaatg ctcttcacta
      301 gacgcgtatt gggatggaac acgcgttcga tacttttagg tacaggcttt tgtgcctact
      361 ggtaagcagt catttgccgg ggcaattaac catctgcact ttattatcag ggtgcagctg
      421 gctaatgatg ctcagtgcca cggggcagga tcagcttctc atctgcacag gaactgtatt
      481 cacagcaaag gacattgacc tttaacaatg aatctttaaa tttaaccggg tcccccagag
      541 atcacctcga tacttttagg tacaggcttt tgtgcctact ggtaagcagt catttgccgg
      601 ggcaattaac catctgcact ttattatcag ggtgcagctg gctaatgatg ctcagtgcca
      661 cggggcagga tcagcttctc atctgcacag gaactgtatt cacagcaaag gacattgacc
      721 tttaacaatg aatctttaaa tttaaccggg tcccccagag atcacctcga tacttttagg
      781 tacaggcttt tgtgcctact ggtaagcagt catttgccgg ggcaattaac catctgcact
      841 ttattatcag ggtgcagctg gctaatgatg ctcagtgcca cggggcagga tcagcttctc
      901 atctgcacag gaactgtatt cacagcaaag gacattgacc tttaacaatg aatctttaaa
      961 tttaaccggg tcccccagag atcaccgagc tcgattcagc cgggagctta gggaggggag
     1021 gtcacttcat aagggcttgg ggggggagtt ggagccacga gtcgtccagc cggagccccg
     1081 tgtggctgtg ctccggcctc agaagcatcc ccggatccag atctttcgaa gctagcgcta
     1141 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     1201 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     1261 gatgccacct acggcaagct gaccctgaag ctgatctgca ccaccggcaa gctgcccgtg
     1321 ccctggccca ccctcgtgac caccctgggc tacggcgtgc agtgcttcgc ccgctacccc
     1381 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     1441 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     1501 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1561 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat caccgccgac
     1621 aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcggc
     1681 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1741 cccgacaacc actacctgag ctaccagtcc aagctgagca aagaccccaa cgagaagcgc
     1801 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1861 ctgtacaagt aagtcgacgg cgcgccgcgg ccgcgaattc gatatcataa tcaacctctg
     1921 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     1981 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     2041 ttctcctcct tgtataaatc ctggttagtt cttgccacgg cggaactcat cgccgcctgc
     2101 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggctcgagag
     2161 atcttcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     2221 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     2281 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     2341 ggaggattgg gaagacaata gcaggcatga gatctcacgt gcggaccgag cggccgcagg
     2401 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
     2461 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag
     2521 cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     2581 tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc
     2641 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc
     2701 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     2761 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     2821 acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     2881 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     2941 caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg
     3001 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt
     3061 tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     3121 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     3181 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     3241 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     3301 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     3361 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3421 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     3481 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     3541 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     3601 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     3661 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     3721 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     3781 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     3841 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     3901 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     3961 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     4021 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     4081 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     4141 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     4201 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     4261 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     4321 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     4381 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     4441 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     4501 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     4561 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     4621 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4681 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4741 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4801 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4861 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4921 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     4981 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     5041 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     5101 gttcctggcc ttttgctggc cttttgctca catgt
//
