LOCUS       sequence-470565-        4966 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0670m 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 4966)
  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 4966)
  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: AiP12878 - pAAV-AiE0670m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4966
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   217..346
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(408..996)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(488..507)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1167..1958,1959..2027))
                     /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     1790..1809
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2028..2132)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2200..2218
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2256..2278)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2378..2397
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2414..2869)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2551..2572)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2763..2782
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2944..3084
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2955..3084
                     /label=AAV2 ITR
     polyA_signal    3202..3313
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3500..3520
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3613..4323,4324..4326,4327..4329))
                     /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(3655..3676)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4004..4023
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4263..4284
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4326..4335
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 catttttatt ttgtttaaac cctgaattat acgtaataca acagctcatt gacaattata
       61 tagcttaatc tatacattca ggaaatatac tacagcatac tctgaaggta ttcattcctt
      121 cctgtaagtc catgtactta tttaattatt tatacatagt tctaaaattt atattgttca
      181 tttgcataag tcaagttgtc ctacgcgtgc ggccgcagga acccctagtg atggagttgg
      241 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
      301 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggacat gtgagcaaaa
      361 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
      421 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
      481 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
      541 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
      601 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
      661 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
      721 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
      781 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
      841 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
      901 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
      961 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     1021 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     1081 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     1141 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     1201 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     1261 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     1321 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     1381 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     1441 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     1501 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     1561 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     1621 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     1681 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     1741 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     1801 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     1861 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     1921 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     1981 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     2041 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     2101 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     2161 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     2221 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     2281 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     2341 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     2401 agagtgcacc ataaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt
     2461 taaatcagct cattttttaa ccaatagacc gaaatcggca aaatccctta taaatcaaaa
     2521 gaatagcccg agatagagtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     2581 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     2641 gaaccatcac ccaaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     2701 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     2761 gaagggaaga aagcgaaagg agcgggcgct aaggcgctgg caagtgtagc ggtcacgctg
     2821 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgta ctatggttgc
     2881 tttgacgtat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
     2941 gcccctgcag gcagctgcgc gctcgctcgc tcactgaggc cgcccgggca aagcccgggc
     3001 gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg
     3061 ccaactccat cactaggggt tcctgcggcc gctcggtccg cacgtgagat ctcatgcctg
     3121 ctattgtctt cccaatcctc ccccttgctg tcctgcccca ccccaccccc cagaatagaa
     3181 tgacacctac tcagacaatg cgatgcaatt tcctcatttt attaggaaag gacagtggga
     3241 gtggcacctt ccagggtcaa ggaaggcacg ggggaggggc aaacaacaga tggctggcaa
     3301 ctagaaggca cagtcgaaga tctctcgagc cacggaattg tcagtgccca acagccgagc
     3361 ccctgtccag cagcgggcaa ggcaggcggc gatgagttcc gccgtggcaa gaactaacca
     3421 ggatttatac aaggaggaga aaatgaaagc catacgggaa gcaatagcat gatacaaagg
     3481 cattaaagca gcgtatccac atagcgtaaa aggagcaaca tagttaagaa taccagtcaa
     3541 tctttcacaa attttgtaat ccagaggttg attatgatat cgaattcgcg gccgcggcgc
     3601 gccgtcgact tacttgtaca gctcgtccat gccgagagtg atcccggcgg cggtcacgaa
     3661 ctccagcagg accatgtgat cgcgcttctc gttggggtct ttgctcagct tggactggta
     3721 gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg tgttctgctg
     3781 gtagtggtcg gcgagctgca cgccgccgtc ctcgatgttg tggcggatct tgaagttggc
     3841 cttgatgccg ttcttctgct tgtcggcggt gatatagacg ttgtggctgt tgtagttgta
     3901 ctccagcttg tgccccagga tgttgccgtc ctccttgaag tcgatgccct tcagctcgat
     3961 gcggttcacc agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt agttgccgtc
     4021 gtccttgaag aagatggtgc gctcctggac gtagccttcg ggcatggcgg acttgaagaa
     4081 gtcgtgctgc ttcatgtggt cggggtagcg ggcgaagcac tgcacgccgt agcccagggt
     4141 ggtcacgagg gtgggccagg gcacgggcag cttgccggtg gtgcagatca gcttcagggt
     4201 cagcttgccg taggtggcat cgccctcgcc ctcgccggac acgctgaact tgtggccgtt
     4261 tacgtcgccg tccagctcga ccaggatggg caccaccccg gtgaacagct cctcgccctt
     4321 gctcaccatg gtggcgaccg gtagcgctag cttcgaaaga tctggatccc agaagcaaat
     4381 gtaagcaata gatggctctg ccctgacttt tatgcccagc ccgagctcgt gttgatcaca
     4441 gcagcataaa taaaaataga acatctgttt ccactttaat ttcctattag tgcgtaagtc
     4501 aatggttttc attaagaata cttacaggat ttgggcattt atagaaagca ttgtgacaaa
     4561 ccagtggtga catcactgaa aatattctat ctccttgtac ataaaattac cattcaaatt
     4621 aactgcctac atgctcccag ggagggcaga agtttcatga gccattcttg gctccataat
     4681 gaaatgtaga tggatacaat cttctgaagg tctgtgcagc tgttaggtca agaccgcagt
     4741 agtcatgcgt tgccttggag caacttacaa gagcatgcta ctctgtacat tccctctccc
     4801 cctgtatcaa catgttctct gagcctcaca gagatttttt tctacttaaa aataaacatt
     4861 tacaggtcaa ttattttcat cactttgagc gggtatgaat cattataaaa agtgtgggat
     4921 tatcattgat tcaaaaatac atgcatttgc ataaaacata tgcaca
//
