LOCUS       sequence-462734-        5375 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2122_3xC2 is an enhancer sequence, designed to drive AAV-mediated
            transgene expression in cortical cell types (note CB bergmann glia).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5375)
  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 5375)
  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: AiP14557 - pAAV-AiE2122_3xC2-minBG-SYFP2-WPRE3...
FEATURES             Location/Qualifiers
     source          1..5375
                     /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      1387..1396
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1393..1395,1396..1398,1399..2109)
                     /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(1438..1459)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1699..1718)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2046..2067
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2202..2222)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2409..2520
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2638..2778
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2638..2767
                     /label=AAV2 ITR
     rep_origin      2853..3308
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2940..2959)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3150..3171
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3325..3344)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3444..3466
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3504..3522)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3590..3694
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3695..3763,3764..4555)
                     /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(3913..3932)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4726..5314
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5215..5234
                     /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 gcgttcctct gtcccactgg ctgtggctca gtgtttattt
      181 attggctgtt tagtatggag aaacatccag tctttcaaca ttgacaacaa atccaaataa
      241 tgagcctgcc catcacgcta cactgggtat catctttgcc tgttaggctc ttggctggtt
      301 ttttgctgta tttgactatc ctcaaagttt gggagttctg actgcgggtt gttattttcc
      361 aattcttatt aacacttgcc aaatcttttg tcatgcatct ttatactcat gacagcttct
      421 aggtcaaatt ttagttttga tcactggatc tttagaagga aagctaagga actgacagac
      481 atagccagct ccttttttta tgctggcaat ggatactgat tagctgttcc tctgtcccac
      541 tggctgtggc tcagtgttta tttattggct gtttagtatg gagaaacatc cagtctttca
      601 acattgacaa caaatccaaa taatgagcct gcccatcacg ctacactggg tatcatcttt
      661 gcctgttagg ctcttggctg gttttttgct gtatttgact atcctcaaag tttgggagtt
      721 ctgactgcgg gttgttattt tccaattctt attaacactt gccaaatctt ttgtcatgca
      781 tctttatact catgacagct tctaggtcaa attttagttt tgatcactgg atctttagaa
      841 ggaaagctaa ggaactgaca gacatagcca gctccttttt ttatgctggc aatggatact
      901 gattagctgt tcctctgtcc cactggctgt ggctcagtgt ttatttattg gctgtttagt
      961 atggagaaac atccagtctt tcaacattga caacaaatcc aaataatgag cctgcccatc
     1021 acgctacact gggtatcatc tttgcctgtt aggctcttgg ctggtttttt gctgtatttg
     1081 actatcctca aagtttggga gttctgactg cgggttgtta ttttccaatt cttattaaca
     1141 cttgccaaat cttttgtcat gcatctttat actcatgaca gcttctaggt caaattttag
     1201 ttttgatcac tggatcttta gaaggaaagc taaggaactg acagacatag ccagctcctt
     1261 tttttatgct ggcaatggat actgattagc tgtgagctcg ggctgggcat aaaagtcagg
     1321 gcagagccat ctattgctta catttgcttc tgggatccag atctttcgaa gctagcgcta
     1381 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     1441 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     1501 gatgccacct acggcaagct gaccctgaag ctgatctgca ccaccggcaa gctgcccgtg
     1561 ccctggccca ccctcgtgac caccctgggc tacggcgtgc agtgcttcgc ccgctacccc
     1621 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     1681 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     1741 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1801 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat caccgccgac
     1861 aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcggc
     1921 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1981 cccgacaacc actacctgag ctaccagtcc aagctgagca aagaccccaa cgagaagcgc
     2041 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     2101 ctgtacaagt aagtcgacgg cgcgccgcgg ccgcgaattc gatatcataa tcaacctctg
     2161 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     2221 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     2281 ttctcctcct tgtataaatc ctggttagtt cttgccacgg cggaactcat cgccgcctgc
     2341 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggctcgagag
     2401 atcttcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     2461 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     2521 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     2581 ggaggattgg gaagacaata gcaggcatga gatctcacgt gcggaccgag cggccgcagg
     2641 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
     2701 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag
     2761 cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     2821 tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc
     2881 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc
     2941 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     3001 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     3061 acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3121 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3181 caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg
     3241 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt
     3301 tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     3361 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     3421 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     3481 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     3541 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     3601 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3661 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     3721 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     3781 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     3841 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     3901 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     3961 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     4021 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     4081 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     4141 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     4201 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     4261 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     4321 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     4381 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     4441 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     4501 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     4561 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     4621 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     4681 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     4741 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     4801 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     4861 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4921 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4981 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5041 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5101 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     5161 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     5221 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     5281 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     5341 gttcctggcc ttttgctggc cttttgctca catgt
//
