LOCUS       sequence-470477-        4666 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0766m 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 4666)
  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 4666)
  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: AiP12623 - pAAV-AiE0766m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4666
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   67..196
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(258..846)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(338..357)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1017..1808,1809..1877))
                     /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     1640..1659
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1878..1982)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2050..2068
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2106..2128)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2228..2247
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2264..2719)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2401..2422)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2613..2632
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2794..2934
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2805..2934
                     /label=AAV2 ITR
     polyA_signal    3052..3163
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3350..3370
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3463..4173,4174..4176,4177..4179))
                     /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(3505..3526)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3854..3873
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4113..4134
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4176..4185
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 agttgatttt aaggtgatat atctcaattt agccactcca caaatacatg ttacgcgtgc
       61 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
      121 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggcgg cctcagtgag cgagcgagcg
      181 cgcagctgcc tgcaggacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
      241 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
      301 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
      361 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
      421 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
      481 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
      541 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
      601 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
      661 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
      721 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
      781 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
      841 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
      901 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
      961 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     1021 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     1081 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     1141 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     1201 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     1261 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     1321 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     1381 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     1441 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     1501 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     1561 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     1621 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     1681 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     1741 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     1801 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     1861 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     1921 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     1981 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     2041 acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt
     2101 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     2161 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
     2221 aactatgcgg catcagagca gattgtactg agagtgcacc ataaaattgt aaacgttaat
     2281 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaatagacc
     2341 gaaatcggca aaatccctta taaatcaaaa gaatagcccg agatagagtt gagtgttgtt
     2401 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     2461 accgtctatc agggcgatgg cccactacgt gaaccatcac ccaaatcaag ttttttgggg
     2521 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     2581 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     2641 aaggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     2701 gcgccgctac agggcgcgta ctatggttgc tttgacgtat gcggtgtgaa ataccgcaca
     2761 gatgcgtaag gagaaaatac cgcatcaggc gcccctgcag gcagctgcgc gctcgctcgc
     2821 tcactgaggc cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag
     2881 tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt tcctgcggcc
     2941 gctcggtccg cacgtgagat ctcatgcctg ctattgtctt cccaatcctc ccccttgctg
     3001 tcctgcccca ccccaccccc cagaatagaa tgacacctac tcagacaatg cgatgcaatt
     3061 tcctcatttt attaggaaag gacagtggga gtggcacctt ccagggtcaa ggaaggcacg
     3121 ggggaggggc aaacaacaga tggctggcaa ctagaaggca cagtcgaaga tctctcgagc
     3181 cacggaattg tcagtgccca acagccgagc ccctgtccag cagcgggcaa ggcaggcggc
     3241 gatgagttcc gccgtggcaa gaactaacca ggatttatac aaggaggaga aaatgaaagc
     3301 catacgggaa gcaatagcat gatacaaagg cattaaagca gcgtatccac atagcgtaaa
     3361 aggagcaaca tagttaagaa taccagtcaa tctttcacaa attttgtaat ccagaggttg
     3421 attatgatat cgaattcgcg gccgcggcgc gccgtcgact tacttgtaca gctcgtccat
     3481 gccgagagtg atcccggcgg cggtcacgaa ctccagcagg accatgtgat cgcgcttctc
     3541 gttggggtct ttgctcagct tggactggta gctcaggtag tggttgtcgg gcagcagcac
     3601 ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg gcgagctgca cgccgccgtc
     3661 ctcgatgttg tggcggatct tgaagttggc cttgatgccg ttcttctgct tgtcggcggt
     3721 gatatagacg ttgtggctgt tgtagttgta ctccagcttg tgccccagga tgttgccgtc
     3781 ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt
     3841 cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtgc gctcctggac
     3901 gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg
     3961 ggcgaagcac tgcacgccgt agcccagggt ggtcacgagg gtgggccagg gcacgggcag
     4021 cttgccggtg gtgcagatca gcttcagggt cagcttgccg taggtggcat cgccctcgcc
     4081 ctcgccggac acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg
     4141 caccaccccg gtgaacagct cctcgccctt gctcaccatg gtggcgaccg gtagcgctag
     4201 cttcgaaaga tctggatccc agaagcaaat gtaagcaata gatggctctg ccctgacttt
     4261 tatgcccagc ccgagctcat gaaaccatgc catcccctgc taccctgagg ctgcagagag
     4321 gtcgcagagg taaagtttcc accgtctgcc tgtgctgaca gagtcatgct cttctcagca
     4381 tggcatttat cctggaggca aaggcacagc agcacaccca ggaaatctcc agggagctga
     4441 aagccatacc atataagata agcactatgc tcaagaaatc agactgattc ttaaactgaa
     4501 aataactagt attttgaatt gaaaatgact cgcatgtggg aagagcttgg ctaaagaaca
     4561 caagactcca gttacacagg aagtgtaaga tctgggctct actgcactgc atggcgactg
     4621 tggttaggaa caaggtctta cttactgtgc tcttcaaatt gctggg
//
