LOCUS       sequence-461914-        5030 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0468m_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 5030)
  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 5030)
  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: AiP13035 -
            pAAV-AiE0468m_3xC2-minBG-SYFP2-WPRE...
FEATURES             Location/Qualifiers
     source          1..5030
                     /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      1042..1051
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1048..1050,1051..1053,1054..1764)
                     /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(1093..1114)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1354..1373)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1701..1722
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1857..1877)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2064..2175
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2293..2433
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2293..2422
                     /label=AAV2 ITR
     rep_origin      2508..2963
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2595..2614)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2805..2826
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2980..2999)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3099..3121
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3159..3177)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3245..3349
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3350..3418,3419..4210)
                     /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(3568..3587)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4381..4969
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4870..4889
                     /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 gcgtgagaat atcaaaccca tcaaaaatca ccgagatcag
      181 tgcaaagact gaagcagaaa gacacagcct ctaaatagga gcggccttaa tgaaagtgca
      241 tcacctcaga gccaacatga agtctgcaga tgggatgcac ctgcctcctg agaacctcac
      301 agagggaaat caagcatcag aacccagaat ggaagcagaa aacccccaat cagactcctc
      361 ggagacactt cagaaaagga actctcagct tgcaaaacaa caggtgcatc tcgagaatat
      421 caaacccatc aaaaatcacc gagatcagtg caaagactga agcagaaaga cacagcctct
      481 aaataggagc ggccttaatg aaagtgcatc acctcagagc caacatgaag tctgcagatg
      541 ggatgcacct gcctcctgag aacctcacag agggaaatca agcatcagaa cccagaatgg
      601 aagcagaaaa cccccaatca gactcctcgg agacacttca gaaaaggaac tctcagcttg
      661 caaaacaaca ggtgcatctc gagaatatca aacccatcaa aaatcaccga gatcagtgca
      721 aagactgaag cagaaagaca cagcctctaa ataggagcgg ccttaatgaa agtgcatcac
      781 ctcagagcca acatgaagtc tgcagatggg atgcacctgc ctcctgagaa cctcacagag
      841 ggaaatcaag catcagaacc cagaatggaa gcagaaaacc cccaatcaga ctcctcggag
      901 acacttcaga aaaggaactc tcagcttgca aaacaacagg tgcatctcga gctcgggctg
      961 ggcataaaag tcagggcaga gccatctatt gcttacattt gcttctggga tccagatctt
     1021 tcgaagctag cgctaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg
     1081 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     1141 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagctgat ctgcaccacc
     1201 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgggctacgg cgtgcagtgc
     1261 ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     1321 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     1381 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     1441 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     1501 tatatcaccg ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac
     1561 atcgaggacg gcggcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     1621 ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccaagct gagcaaagac
     1681 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     1741 ctcggcatgg acgagctgta caagtaagtc gacggcgcgc cgcggccgcg aattcgatat
     1801 cataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     1861 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     1921 cgtatggctt tcattttctc ctccttgtat aaatcctggt tagttcttgc cacggcggaa
     1981 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     2041 tccgtggctc gagagatctt cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     2101 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     2161 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     2221 gcaggacagc aagggggagg attgggaaga caatagcagg catgagatct cacgtgcgga
     2281 ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     2341 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     2401 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta
     2461 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag
     2521 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     2581 cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     2641 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     2701 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata
     2761 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     2821 aactggaaca acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc
     2881 gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     2941 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc
     3001 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     3061 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     3121 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     3181 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     3241 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     3301 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     3361 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     3421 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     3481 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     3541 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     3601 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     3661 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     3721 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     3781 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     3841 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     3901 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     3961 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     4021 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     4081 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     4141 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     4201 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     4261 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     4321 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     4381 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     4441 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     4501 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     4561 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     4621 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     4681 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     4741 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     4801 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     4861 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     4921 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     4981 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
