LOCUS       sequence-462398-        5765 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0078m_3xC2 is an enhancer sequence, designed to drive 
            AAV-mediated transgene expression in specific populations of brain 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5765)
  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 5765)
  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: AiP11955 -
            pAAV-hsA2-AiE0078m_3xC2-minRho-SYFP...
FEATURES             Location/Qualifiers
     source          1..5765
                     /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      1802..1811
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1808..1810,1811..1813,1814..2524)
                     /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(1853..1874)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2114..2133)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2461..2482
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2592..2612)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2799..2910
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3028..3168
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3028..3157
                     /label=AAV2 ITR
     rep_origin      3243..3698
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3330..3349)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3540..3561
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3715..3734)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3834..3856
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3894..3912)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3980..4084
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4085..4153,4154..4945)
                     /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(4303..4322)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5116..5704
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5605..5624
                     /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 gcatggtcca gcaaagcctt tctgttctaa aggaaaggat ctgagttgtc acctcccagg
      421 tccgtggaag gctttttagc agtttggcag gtgcctgagg gccacacctc atctgcataa
      481 aatgtggcag attgcaaccg ccctcgtctt ttttattttt aaactggttt ttgaaacaga
      541 acatatataa aagctcagag aaagggaaag gagatagatg gccgagcttc catatccctt
      601 agtgccttta ttttcattct ttttccattt tcctaagtgg ctatttacca agacaaagat
      661 aacaaatctg ctaggaaaag gagtgggcag tgctacaaaa tgtttttttt ttttaaagaa
      721 agtcctatct tataatagat cttcaccacg atgcctcgag cagagccctc atcacacaga
      781 ctgaagcatg gtccagcaaa gcctttctgt tctaaaggaa aggatctgag ttgtcacctc
      841 ccaggtccgt ggaaggcttt ttagcagttt ggcaggtgcc tgagggccac acctcatctg
      901 cataaaatgt ggcagattgc aaccgccctc gtctttttta tttttaaact ggtttttgaa
      961 acagaacata tataaaagct cagagaaagg gaaaggagat agatggccga gcttccatat
     1021 cccttagtgc ctttattttc attctttttc cattttccta agtggctatt taccaagaca
     1081 aagataacaa atctgctagg aaaaggagtg ggcagtgcta caaaatgttt tttttttttt
     1141 aaagaaagtc ctatcttata atagatcttc accacgatgc ctctgggtga gtaagagtag
     1201 gtggcaacga agcatggtcc agcaaagcct ttctgttcta aaggaaagga tctgagttgt
     1261 cacctcccag gtccgtggaa ggctttttag cagtttggca ggtgcctgag ggccacacct
     1321 catctgcata aaatgtggca gattgcaacc gccctcgtct tttttatttt taaactggtt
     1381 tttgaaacag aacatatata aaagctcaga gaaagggaaa ggagatagat ggccgagctt
     1441 ccatatccct tagtgccttt attttcattc tttttccatt ttcctaagtg gctatttacc
     1501 aagacaaaga taacaaatct gctaggaaaa ggagtgggca gtgctacaaa atgttttttt
     1561 ttttttaaag aaagtcctat cttataatag atcttcacca cgatgcctcg aattcgatat
     1621 cataatcaac cataggtacc gagctcggga ttcagccggg agcttaggga ggggaggtca
     1681 cttcataagg gcctgggggg ggagttggag ccacgagtcg tccagccgga gccccgtgtg
     1741 gctgagctcc ggcctcagaa gcatccccgg gttggatcct tcgaagctag cgctaccggt
     1801 cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     1861 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     1921 cacctacggc aagctgaccc tgaagctgat ctgcaccacc ggcaagctgc ccgtgccctg
     1981 gcccaccctc gtgaccaccc tgggctacgg cgtgcagtgc ttcgcccgct accccgacca
     2041 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     2101 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     2161 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     2221 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcaccg ccgacaagca
     2281 gaagaacggc atcaaggcca acttcaagat ccgccacaac atcgaggacg gcggcgtgca
     2341 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     2401 caaccactac ctgagctacc agtccaagct gagcaaagac cccaacgaga agcgcgatca
     2461 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     2521 caagtaagtc gacatcataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     2581 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     2641 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttagtt
     2701 cttgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     2761 ttgggcactg acaattccgt ggctcgagag atcttcgact gtgccttcta gttgccagcc
     2821 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     2881 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     2941 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatga
     3001 gatctcacgt gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct
     3061 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct
     3121 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc
     3181 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata
     3241 gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     3301 cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     3361 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     3421 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg
     3481 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     3541 tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt
     3601 ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt
     3661 taacgggaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat
     3721 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     3781 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     3841 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     3901 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     3961 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     4021 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     4081 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     4141 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     4201 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     4261 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     4321 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     4381 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     4441 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     4501 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     4561 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     4621 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     4681 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     4741 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     4801 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     4861 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     4921 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     4981 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     5041 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     5101 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5161 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5221 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     5281 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5341 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     5401 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     5461 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     5521 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     5581 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     5641 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     5701 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     5761 catgt
//
