LOCUS       sequence-449740-        5225 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0370m_3xC2 is an enhancer sequence, designed to drive 
            AAV-mediated transgene expression in cortical L5_ET neurons (VISp 
            enriched).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5225)
  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 5225)
  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: AiP14474 -
            pAAV-AiE0370m_3xC2-minBG-SYFP2-WPRE...
FEATURES             Location/Qualifiers
     source          1..5225
                     /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      1237..1246
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1243..1245,1246..1248,1249..1959)
                     /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(1288..1309)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1549..1568)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1896..1917
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2052..2072)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2259..2370
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2488..2628
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2488..2617
                     /label=AAV2 ITR
     rep_origin      2703..3158
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2790..2809)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3000..3021
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3175..3194)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3294..3316
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3354..3372)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3440..3544
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3545..3613,3614..4405)
                     /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(3763..3782)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4576..5164
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5065..5084
                     /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 gcgtacaagt gagaaagaga aatggagatt ttaccattta
      181 atcatctccg aagagaaggt gagaagaaat tgagaaaggt aatttgcata ccatgcatag
      241 gcattagaga aagcattttt tttcttaact ctccatttgt ttacagatat ttccatatgg
      301 ccttttaaaa acttcaacag agaaattcga acacaactaa ctggaatttc aattttcata
      361 taaaaccagc atcatagata tatatgtagc tactctccca cttcacatta ttcagttagt
      421 atattttcta agctaatttc caatccattt taagactatg tacagctcat atgaggcaca
      481 agtgagaaag agaaatggag attttaccat ttaatcatct ccgaagagaa ggtgagaaga
      541 aattgagaaa ggtaatttgc ataccatgca taggcattag agaaagcatt ttttttctta
      601 actctccatt tgtttacaga tatttccata tggcctttta aaaacttcaa cagagaaatt
      661 cgaacacaac taactggaat ttcaattttc atataaaacc agcatcatag atatatatgt
      721 agctactctc ccacttcaca ttattcagtt agtatatttt ctaagctaat ttccaatcca
      781 ttttaagact atgtacagct catatgaggc acaagtgaga aagagaaatg gagattttac
      841 catttaatca tctccgaaga gaaggtgaga agaaattgag aaaggtaatt tgcataccat
      901 gcataggcat tagagaaagc attttttttc ttaactctcc atttgtttac agatatttcc
      961 atatggcctt ttaaaaactt caacagagaa attcgaacac aactaactgg aatttcaatt
     1021 ttcatataaa accagcatca tagatatata tgtagctact ctcccacttc acattattca
     1081 gttagtatat tttctaagct aatttccaat ccattttaag actatgtaca gctcatatga
     1141 ggcgagctcg ggctgggcat aaaagtcagg gcagagccat ctattgctta catttgcttc
     1201 tgggatccag atctttcgaa gctagcgcta ccggtcgcca ccatggtgag caagggcgag
     1261 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     1321 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     1381 ctgatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgggc
     1441 tacggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag
     1501 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     1561 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     1621 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     1681 aacagccaca acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc
     1741 aagatccgcc acaacatcga ggacggcggc gtgcagctcg ccgaccacta ccagcagaac
     1801 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc
     1861 aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     1921 gccgccggga tcactctcgg catggacgag ctgtacaagt aagtcgacgg cgcgccgcgg
     1981 ccgcgaattc gatatcataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     2041 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     2101 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttagtt
     2161 cttgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     2221 ttgggcactg acaattccgt ggctcgagag atcttcgact gtgccttcta gttgccagcc
     2281 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     2341 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     2401 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatga
     2461 gatctcacgt gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct
     2521 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct
     2581 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc
     2641 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata
     2701 gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     2761 cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     2821 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     2881 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg
     2941 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     3001 tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt
     3061 ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt
     3121 taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat
     3181 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     3241 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     3301 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     3361 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     3421 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     3481 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     3541 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     3601 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     3661 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     3721 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     3781 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     3841 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     3901 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     3961 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     4021 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     4081 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     4141 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     4201 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     4261 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     4321 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     4381 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     4441 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     4501 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     4561 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     4621 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     4681 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     4741 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     4801 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     4861 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     4921 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     4981 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     5041 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     5101 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     5161 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     5221 catgt
//
