LOCUS       sequence-449788-        4735 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0671m_3xC2 is an enhancer sequence, designed to drive 
            AAV-mediated transgene expression in cortical L4/5_IT neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4735)
  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 4735)
  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: AiP13897 -
            pAAV-AiE0671m_3xC2-minBG-SYFP2-WPRE...
FEATURES             Location/Qualifiers
     source          1..4735
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     regulatory      747..756
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(753..755,756..758,759..1469)
                     /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(798..819)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1059..1078)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1406..1427
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1562..1582)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1769..1880
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1998..2138
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1998..2127
                     /label=AAV2 ITR
     rep_origin      2213..2668
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2300..2319)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2510..2531
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2685..2704)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2804..2826
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2864..2882)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2950..3054
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3055..3123,3124..3915)
                     /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(3273..3292)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4086..4674
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4575..4594
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtaaagg ttgcactgta tctgaaataa
      181 gctacatttc cttcccagct caccaaaaag caggacatat ggatgtgaac aagagatcca
      241 ggacagctga agtactttta aggaacactg taaactttcg tctctaaaat taatcatctc
      301 tatgtatgag tatatgttca gaaaggttgc actgtatctg aaataagcta catttccttc
      361 ccagctcacc aaaaagcagg acatatggat gtgaacaaga gatccaggac agctgaagta
      421 cttttaagga acactgtaaa ctttcgtctc taaaattaat catctctatg tatgagtata
      481 tgttcagaaa ggttgcactg tatctgaaat aagctacatt tccttcccag ctcaccaaaa
      541 agcaggacat atggatgtga acaagagatc caggacagct gaagtacttt taaggaacac
      601 tgtaaacttt cgtctctaaa attaatcatc tctatgtatg agtatatgtt caggagctcg
      661 ggctgggcat aaaagtcagg gcagagccat ctattgctta catttgcttc tgggatccag
      721 atctttcgaa gctagcgcta ccggtcgcca ccatggtgag caagggcgag gagctgttca
      781 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
      841 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ctgatctgca
      901 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgggc tacggcgtgc
      961 agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     1021 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     1081 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     1141 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     1201 acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc aagatccgcc
     1261 acaacatcga ggacggcggc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     1321 gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc aagctgagca
     1381 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     1441 tcactctcgg catggacgag ctgtacaagt aagtcgacgg cgcgccgcgg ccgcgaattc
     1501 gatatcataa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     1561 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     1621 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttagtt cttgccacgg
     1681 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     1741 acaattccgt ggctcgagag atcttcgact gtgccttcta gttgccagcc atctgttgtt
     1801 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     1861 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     1921 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatga gatctcacgt
     1981 gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc
     2041 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc
     2101 ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct
     2161 ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc
     2221 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     2281 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     2341 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     2401 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     2461 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     2521 ttccaaactg gaacaacact caaccctatc tcgggctatt cttttgattt ataagggatt
     2581 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     2641 tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat
     2701 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     2761 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     2821 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     2881 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     2941 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     3001 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     3061 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     3121 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     3181 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     3241 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     3301 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     3361 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     3421 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     3481 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     3541 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     3601 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     3661 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     3721 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     3781 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     3841 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     3901 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     3961 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     4021 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     4081 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     4141 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     4201 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     4261 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     4321 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     4381 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     4441 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     4501 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     4561 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     4621 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     4681 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgt
//
