LOCUS       sequence-462823-        4847 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0523h_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 4847)
  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 4847)
  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: AiP15469 -
            pAAV-AiE0523h_3xC2-minBG-SYFP2-WPRE...
FEATURES             Location/Qualifiers
     source          1..4847
                     /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      859..868
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(865..867,868..870,871..1581)
                     /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(910..931)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1171..1190)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1518..1539
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1674..1694)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1881..1992
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2110..2250
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2110..2239
                     /label=AAV2 ITR
     rep_origin      2325..2780
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2412..2431)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2622..2643
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2797..2816)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2916..2938
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2976..2994)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3062..3166
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3167..3235,3236..4027)
                     /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(3385..3404)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4198..4786
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4687..4706
                     /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 gcgtgggtgc tagaagaagt tacatttacc caatgcattg
      181 gatattagat atacccaaga taactgtcag gtcaggtggt gtgaagacag ggaagatttt
      241 ggagccagta atcgggaata taatggggga aggaagagaa gccagaaaac aaagactaaa
      301 gtagtggttc ttaactctgg ctgtatatta cagcacatat ggtcctttgg agggtgctag
      361 aagaagttac atttacccaa tgcattggat attagatata cccaagataa ctgtcaggtc
      421 aggtggtgtg aagacaggga agattttgga gccagtaatc gggaatataa tgggggaagg
      481 aagagaagcc agaaaacaaa gactaaagta gtggttctta actctggctg tatattacag
      541 cacatatggt cctttggagg gtgctagaag aagttacatt tacccaatgc attggatatt
      601 agatataccc aagataactg tcaggtcagg tggtgtgaag acagggaaga ttttggagcc
      661 agtaatcggg aatataatgg gggaaggaag agaagccaga aaacaaagac taaagtagtg
      721 gttcttaact ctggctgtat attacagcac atatggtcct ttggagagct cgggctgggc
      781 ataaaagtca gggcagagcc atctattgct tacatttgct tctgggatcc agatctttcg
      841 aagctagcgc taccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg
      901 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
      961 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agctgatctg caccaccggc
     1021 aagctgcccg tgccctggcc caccctcgtg accaccctgg gctacggcgt gcagtgcttc
     1081 gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     1141 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     1201 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     1261 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     1321 atcaccgccg acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacatc
     1381 gaggacggcg gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     1441 cccgtgctgc tgcccgacaa ccactacctg agctaccagt ccaagctgag caaagacccc
     1501 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     1561 ggcatggacg agctgtacaa gtaagtcgac ggcgcgccgc ggccgcgaat tcgatatcat
     1621 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     1681 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     1741 atggctttca ttttctcctc cttgtataaa tcctggttag ttcttgccac ggcggaactc
     1801 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     1861 gtggctcgag agatcttcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     1921 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     1981 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     2041 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gagatctcac gtgcggaccg
     2101 agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
     2161 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag
     2221 tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc
     2281 atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg
     2341 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     2401 cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     2461 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     2521 cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac
     2581 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     2641 tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat
     2701 ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     2761 aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata
     2821 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     2881 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     2941 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     3001 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     3061 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     3121 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     3181 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     3241 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     3301 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     3361 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     3421 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     3481 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     3541 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     3601 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     3661 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     3721 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     3781 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     3841 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     3901 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     3961 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     4021 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     4081 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     4141 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     4201 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     4261 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     4321 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     4381 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4441 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4501 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4561 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4621 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4681 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4741 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     4801 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgt
//
