LOCUS       sequence-470571-        4530 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0668m 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 4530)
  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 4530)
  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: AiP12886 - pAAV-AiE0668m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4530
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   127..256
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(318..906)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(398..417)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1077..1868,1869..1937))
                     /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     1700..1719
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1938..2042)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2110..2128
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2166..2188)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2288..2307
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2324..2779)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2461..2482)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2673..2692
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2854..2994
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2865..2994
                     /label=AAV2 ITR
     polyA_signal    3112..3223
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3410..3430
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3523..4233,4234..4236,4237..4239))
                     /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(3565..3586)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3914..3933
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4173..4194
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4236..4245
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 taaatgtgta tgttttataa ccccaacagt ctttgatgtt gtatttatcc ctttcttagg
       61 tttttaagag ttgtttttgt ttttatttgt gtgtgtgtgt gtgtgtgtgt gtacgcgtgc
      121 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
      181 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggcgg cctcagtgag cgagcgagcg
      241 cgcagctgcc tgcaggacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
      301 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
      361 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
      421 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
      481 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
      541 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
      601 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
      661 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
      721 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
      781 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
      841 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
      901 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
      961 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     1021 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     1081 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     1141 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     1201 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     1261 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     1321 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     1381 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     1441 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     1501 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     1561 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     1621 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     1681 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     1741 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     1801 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     1861 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     1921 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     1981 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     2041 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     2101 acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt
     2161 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     2221 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
     2281 aactatgcgg catcagagca gattgtactg agagtgcacc ataaaattgt aaacgttaat
     2341 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaatagacc
     2401 gaaatcggca aaatccctta taaatcaaaa gaatagcccg agatagagtt gagtgttgtt
     2461 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     2521 accgtctatc agggcgatgg cccactacgt gaaccatcac ccaaatcaag ttttttgggg
     2581 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     2641 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     2701 aaggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     2761 gcgccgctac agggcgcgta ctatggttgc tttgacgtat gcggtgtgaa ataccgcaca
     2821 gatgcgtaag gagaaaatac cgcatcaggc gcccctgcag gcagctgcgc gctcgctcgc
     2881 tcactgaggc cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag
     2941 tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt tcctgcggcc
     3001 gctcggtccg cacgtgagat ctcatgcctg ctattgtctt cccaatcctc ccccttgctg
     3061 tcctgcccca ccccaccccc cagaatagaa tgacacctac tcagacaatg cgatgcaatt
     3121 tcctcatttt attaggaaag gacagtggga gtggcacctt ccagggtcaa ggaaggcacg
     3181 ggggaggggc aaacaacaga tggctggcaa ctagaaggca cagtcgaaga tctctcgagc
     3241 cacggaattg tcagtgccca acagccgagc ccctgtccag cagcgggcaa ggcaggcggc
     3301 gatgagttcc gccgtggcaa gaactaacca ggatttatac aaggaggaga aaatgaaagc
     3361 catacgggaa gcaatagcat gatacaaagg cattaaagca gcgtatccac atagcgtaaa
     3421 aggagcaaca tagttaagaa taccagtcaa tctttcacaa attttgtaat ccagaggttg
     3481 attatgatat cgaattcgcg gccgcggcgc gccgtcgact tacttgtaca gctcgtccat
     3541 gccgagagtg atcccggcgg cggtcacgaa ctccagcagg accatgtgat cgcgcttctc
     3601 gttggggtct ttgctcagct tggactggta gctcaggtag tggttgtcgg gcagcagcac
     3661 ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg gcgagctgca cgccgccgtc
     3721 ctcgatgttg tggcggatct tgaagttggc cttgatgccg ttcttctgct tgtcggcggt
     3781 gatatagacg ttgtggctgt tgtagttgta ctccagcttg tgccccagga tgttgccgtc
     3841 ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt
     3901 cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtgc gctcctggac
     3961 gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg
     4021 ggcgaagcac tgcacgccgt agcccagggt ggtcacgagg gtgggccagg gcacgggcag
     4081 cttgccggtg gtgcagatca gcttcagggt cagcttgccg taggtggcat cgccctcgcc
     4141 ctcgccggac acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg
     4201 caccaccccg gtgaacagct cctcgccctt gctcaccatg gtggcgaccg gtagcgctag
     4261 cttcgaaaga tctggatccc agaagcaaat gtaagcaata gatggctctg ccctgacttt
     4321 tatgcccagc ccgagctcac ctcatgtctt ctgtagtttt caggattgga tgccaacata
     4381 tgttagcttg ggaagcagat cgggaaagct catggtctgg gcttcctttg ctatgctctc
     4441 tagttgttta tgttgcacaa gaataatttg ttctgtgaca ttttggatac tcatttgtat
     4501 gtaattttag tagcatcttg gaaatgtcta
//
