LOCUS       sequence-470568-        4507 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0681m 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 4507)
  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 4507)
  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: AiP12879 - pAAV-AiE0681m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4507
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(257..1048,1049..1117))
                     /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     880..899
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1118..1222)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1290..1308
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1346..1368)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1468..1487
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1504..1959)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1641..1662)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1853..1872
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2034..2174
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2045..2174
                     /label=AAV2 ITR
     polyA_signal    2292..2403
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2590..2610
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(2703..3413,3414..3416,3417..3419))
                     /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(2745..2766)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3094..3113
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3353..3374
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3416..3425
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   3814..3943
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(join(4005..4507,1..86))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4085..4104)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
       61 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
      121 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
      181 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
      241 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
      301 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
      361 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
      421 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
      481 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
      541 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
      601 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
      661 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
      721 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
      781 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
      841 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
      901 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
      961 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     1021 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     1081 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     1141 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     1201 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     1261 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
     1321 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     1381 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     1441 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     1501 ataaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     1561 cattttttaa ccaatagacc gaaatcggca aaatccctta taaatcaaaa gaatagcccg
     1621 agatagagtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     1681 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     1741 ccaaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     1801 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     1861 aagcgaaagg agcgggcgct aaggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     1921 ccacacccgc cgcgcttaat gcgccgctac agggcgcgta ctatggttgc tttgacgtat
     1981 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gcccctgcag
     2041 gcagctgcgc gctcgctcgc tcactgaggc cgcccgggca aagcccgggc gtcgggcgac
     2101 ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat
     2161 cactaggggt tcctgcggcc gctcggtccg cacgtgagat ctcatgcctg ctattgtctt
     2221 cccaatcctc ccccttgctg tcctgcccca ccccaccccc cagaatagaa tgacacctac
     2281 tcagacaatg cgatgcaatt tcctcatttt attaggaaag gacagtggga gtggcacctt
     2341 ccagggtcaa ggaaggcacg ggggaggggc aaacaacaga tggctggcaa ctagaaggca
     2401 cagtcgaaga tctctcgagc cacggaattg tcagtgccca acagccgagc ccctgtccag
     2461 cagcgggcaa ggcaggcggc gatgagttcc gccgtggcaa gaactaacca ggatttatac
     2521 aaggaggaga aaatgaaagc catacgggaa gcaatagcat gatacaaagg cattaaagca
     2581 gcgtatccac atagcgtaaa aggagcaaca tagttaagaa taccagtcaa tctttcacaa
     2641 attttgtaat ccagaggttg attatgatat cgaattcgcg gccgcggcgc gccgtcgact
     2701 tacttgtaca gctcgtccat gccgagagtg atcccggcgg cggtcacgaa ctccagcagg
     2761 accatgtgat cgcgcttctc gttggggtct ttgctcagct tggactggta gctcaggtag
     2821 tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg
     2881 gcgagctgca cgccgccgtc ctcgatgttg tggcggatct tgaagttggc cttgatgccg
     2941 ttcttctgct tgtcggcggt gatatagacg ttgtggctgt tgtagttgta ctccagcttg
     3001 tgccccagga tgttgccgtc ctccttgaag tcgatgccct tcagctcgat gcggttcacc
     3061 agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt agttgccgtc gtccttgaag
     3121 aagatggtgc gctcctggac gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc
     3181 ttcatgtggt cggggtagcg ggcgaagcac tgcacgccgt agcccagggt ggtcacgagg
     3241 gtgggccagg gcacgggcag cttgccggtg gtgcagatca gcttcagggt cagcttgccg
     3301 taggtggcat cgccctcgcc ctcgccggac acgctgaact tgtggccgtt tacgtcgccg
     3361 tccagctcga ccaggatggg caccaccccg gtgaacagct cctcgccctt gctcaccatg
     3421 gtggcgaccg gtagcgctag cttcgaaaga tctggatccc agaagcaaat gtaagcaata
     3481 gatggctctg ccctgacttt tatgcccagc ccgagctctt aggtcccagt ctggcattgt
     3541 catacaatct ccttacatcg gaagggcagc tagagggatg cacagtgaac agagtaacag
     3601 tggctcctac aaggacacga ctgtgaggaa caatgactag actctgtcta gcaaagctca
     3661 caggattctc agtgggcggc aggagcagag caacgggtgt acttctgctg cacaactgag
     3721 cccatgtagt ggctagaagg aaggagagac actcgcctct atttaggcgc agtgcctttc
     3781 aaagaaaaga cttctaagca cgcgtgcggc cgcaggaacc cctagtgatg gagttggcca
     3841 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     3901 cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggacatgtg agcaaaaggc
     3961 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     4021 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     4081 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     4141 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     4201 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     4261 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     4321 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     4381 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     4441 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     4501 ggtagct
//
