LOCUS       sequence-449789-        4915 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0647m_3xC2 is an enhancer sequence, designed to drive 
            AAV-mediated transgene expression in cortical L5_ET neurons (note 
            HPF subiculum pyr; MB dopaminergic).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4915)
  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 4915)
  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: AiP13898 -
            pAAV-AiE0647m_3xC2-minBG-SYFP2-WPRE...
FEATURES             Location/Qualifiers
     source          1..4915
                     /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      927..936
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(933..935,936..938,939..1649)
                     /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(978..999)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1239..1258)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1586..1607
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1742..1762)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1949..2060
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2178..2318
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2178..2307
                     /label=AAV2 ITR
     rep_origin      2393..2848
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2480..2499)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2690..2711
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2865..2884)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2984..3006
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3044..3062)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3130..3234
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3235..3303,3304..4095)
                     /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(3453..3472)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4266..4854
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4755..4774
                     /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 cgcgtatttt taactaaaca aaaaaaatag
      181 gtaatagaac atttcatatt gaactcccag gatagcattt atcataatcc accagttgaa
      241 ttattcatgc gctgataaat aaccttagcc tccggcacag taatcctctc actgaataat
      301 caccagggct ccctgcccgt ggtgttttgc tttacaacat aaattttaaa attattctgt
      361 gttggcatac atatgtgaaa gatttttaac taaacaaaaa aaataggtaa tagaacattt
      421 catattgaac tcccaggata gcatttatca taatccacca gttgaattat tcatgcgctg
      481 ataaataacc ttagcctccg gcacagtaat cctctcactg aataatcacc agggctccct
      541 gcccgtggtg ttttgcttta caacataaat tttaaaatta ttctgtgttg gcatacatat
      601 gtgaaagatt tttaactaaa caaaaaaaat aggtaataga acatttcata ttgaactccc
      661 aggatagcat ttatcataat ccaccagttg aattattcat gcgctgataa ataaccttag
      721 cctccggcac agtaatcctc tcactgaata atcaccaggg ctccctgccc gtggtgtttt
      781 gctttacaac ataaatttta aaattattct gtgttggcat acatatgtga aaggagctcg
      841 ggctgggcat aaaagtcagg gcagagccat ctattgctta catttgcttc tgggatccag
      901 atctttcgaa gctagcgcta ccggtcgcca ccatggtgag caagggcgag gagctgttca
      961 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     1021 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ctgatctgca
     1081 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgggc tacggcgtgc
     1141 agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     1201 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     1261 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     1321 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     1381 acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc aagatccgcc
     1441 acaacatcga ggacggcggc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     1501 gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc aagctgagca
     1561 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     1621 tcactctcgg catggacgag ctgtacaagt aagtcgacgg cgcgccgcgg ccgcgaattc
     1681 gatatcataa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     1741 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     1801 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttagtt cttgccacgg
     1861 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     1921 acaattccgt ggctcgagag atcttcgact gtgccttcta gttgccagcc atctgttgtt
     1981 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     2041 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     2101 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatga gatctcacgt
     2161 gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc
     2221 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc
     2281 ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct
     2341 ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc
     2401 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     2461 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     2521 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     2581 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     2641 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     2701 ttccaaactg gaacaacact caaccctatc tcgggctatt cttttgattt ataagggatt
     2761 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     2821 tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat
     2881 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     2941 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     3001 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     3061 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     3121 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     3181 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     3241 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     3301 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     3361 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     3421 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     3481 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     3541 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     3601 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     3661 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     3721 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     3781 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     3841 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     3901 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     3961 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     4021 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     4081 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     4141 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     4201 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     4261 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     4321 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     4381 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     4441 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     4501 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     4561 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     4621 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     4681 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     4741 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     4801 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     4861 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgt
//
