LOCUS       sequence-466354-        4789 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE3051m 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 4789)
  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 4789)
  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: AiP20042 - pAAV-AiE3051m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4789
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     34..53
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   215..355
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   226..355
                     /label=AAV2 ITR
     polyA_signal    473..584
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     771..791
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(884..1594,1595..1597,1598..1600))
                     /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(926..947)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1275..1294
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1534..1555
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1597..1606
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   2277..2406
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2468..3056)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2548..2567)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3227..4018,4019..4087))
                     /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     3850..3869
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4088..4192)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4260..4278
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4316..4338)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4438..4457
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(join(4474..4789,1..140))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4611..4632)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
       61 taaggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
      121 tgcgccgcta cagggcgcgt actatggttg ctttgacgta tgcggtgtga aataccgcac
      181 agatgcgtaa ggagaaaata ccgcatcagg cgcccctgca ggcagctgcg cgctcgctcg
      241 ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg cccggcctca
      301 gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg ttcctgcggc
      361 cgctcggtcc gcacgtgaga tctcatgcct gctattgtct tcccaatcct cccccttgct
      421 gtcctgcccc accccacccc ccagaataga atgacaccta ctcagacaat gcgatgcaat
      481 ttcctcattt tattaggaaa ggacagtggg agtggcacct tccagggtca aggaaggcac
      541 gggggagggg caaacaacag atggctggca actagaaggc acagtcgaag atctctcgag
      601 ccacggaatt gtcagtgccc aacagccgag cccctgtcca gcagcgggca aggcaggcgg
      661 cgatgagttc cgccgtggca agaactaacc aggatttata caaggaggag aaaatgaaag
      721 ccatacggga agcaatagca tgatacaaag gcattaaagc agcgtatcca catagcgtaa
      781 aaggagcaac atagttaaga ataccagtca atctttcaca aattttgtaa tccagaggtt
      841 gattatgata tcgaattcgc ggccgcggcg cgccgtcgac ttacttgtac agctcgtcca
      901 tgccgagagt gatcccggcg gcggtcacga actccagcag gaccatgtga tcgcgcttct
      961 cgttggggtc tttgctcagc ttggactggt agctcaggta gtggttgtcg ggcagcagca
     1021 cggggccgtc gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc acgccgccgt
     1081 cctcgatgtt gtggcggatc ttgaagttgg ccttgatgcc gttcttctgc ttgtcggcgg
     1141 tgatatagac gttgtggctg ttgtagttgt actccagctt gtgccccagg atgttgccgt
     1201 cctccttgaa gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact
     1261 tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg cgctcctgga
     1321 cgtagccttc gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc
     1381 gggcgaagca ctgcacgccg tagcccaggg tggtcacgag ggtgggccag ggcacgggca
     1441 gcttgccggt ggtgcagatc agcttcaggg tcagcttgcc gtaggtggca tcgccctcgc
     1501 cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg
     1561 gcaccacccc ggtgaacagc tcctcgccct tgctcaccat ggtggcgacc ggtagcgcta
     1621 gcttcgaaag atctggatcc cagaagcaaa tgtaagcaat agatggctct gccctgactt
     1681 ttatgcccag cccgagctcc gccggcgctg cccagagttt atggtgtgat tttcttttct
     1741 taccaagaag ctttcagaaa aataggtttt gaaccatatt ctccatttca cctaagaaat
     1801 caatgttagc ctaaggagac taggaattga gagggtaaat acagacaaga ttatattccg
     1861 tcccaggctt atatctacaa tgtggtcaga aatgatggag agataacggg tagggggcgt
     1921 gatgccactc agttttctgt aaagagcttc cttgatctgt tcagcccaag ggggacttac
     1981 ctggtaagtc ccaaagcctt tctagaacta cagcaggaga gagcaggttt catgacatag
     2041 agctggcagg tgcacagact atcaggctag acagatagtc actgacactt ctaagcctaa
     2101 tttagtcgtc tccacagaag cagcctctcc tagctgatgc attttatttt tctagggtct
     2161 gaagatggat caaggactta agtgctggtt accttgaggg aattcagagc ataggtgacc
     2221 ctgctgtgct ggggttgaag tcatggttgt tctagtgaat aaacgcgtgc ggccgcagga
     2281 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     2341 gcgaccaaag gtcgcccgac gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     2401 tgcaggacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     2461 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     2521 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     2581 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     2641 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     2701 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     2761 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     2821 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     2881 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     2941 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     3001 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     3061 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     3121 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     3181 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     3241 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     3301 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     3361 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     3421 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     3481 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     3541 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     3601 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     3661 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     3721 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     3781 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     3841 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     3901 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     3961 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     4021 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     4081 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     4141 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     4201 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     4261 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     4321 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     4381 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     4441 catcagagca gattgtactg agagtgcacc ataaaattgt aaacgttaat attttgttaa
     4501 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaatagacc gaaatcggca
     4561 aaatccctta taaatcaaaa gaatagcccg agatagagtt gagtgttgtt ccagtttgga
     4621 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     4681 agggcgatgg cccactacgt gaaccatcac ccaaatcaag ttttttgggg tcgaggtgcc
     4741 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttg
//
